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Search Results (507)

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38 pages, 4078 KiB  
Article
Morphological, Toxicological, and Biochemical Characterization of Two Species of Gambierdiscus from Bahía de La Paz, Gulf of California
by Leyberth José Fernández-Herrera, Erick Julián Núñez-Vázquez, Francisco E. Hernández-Sandoval, Daniel Octavio Ceseña-Ojeda, Sara García-Davis, Andressa Teles, Marte Virgen-Félix and Dariel Tovar-Ramírez
Mar. Drugs 2024, 22(9), 422; https://doi.org/10.3390/md22090422 - 16 Sep 2024
Abstract
We describe five new isolates of two Gambierdiscus species from Bahía de La Paz in the southern Gulf of California. Batch cultures of Gambierdiscus were established for morphological characterization using light microscopy (LM) and scanning electron microscopy (SEM). Pigment and amino acid profiles [...] Read more.
We describe five new isolates of two Gambierdiscus species from Bahía de La Paz in the southern Gulf of California. Batch cultures of Gambierdiscus were established for morphological characterization using light microscopy (LM) and scanning electron microscopy (SEM). Pigment and amino acid profiles were also analyzed using high-performance liquid chromatography (HPLC-UV and HPLC-DAD). Finally, toxicity (CTX-like and MTX-like activity) was evaluated using the Artemia salina assay (ARTOX), mouse assay (MBA), marine fish assay (MFA), and fluorescent receptor binding assay (fRBA). These strains were identified as Gambierdiscus cf. caribaeus and Gambierdiscus cf. carpenteri. Toxicity for CTX-like and MTX-like activity was confirmed in all evaluated clones. Seven pigments were detected, with chlorophyll a, pyridine, Chl2, and diadinoxanthin being particularly noteworthy. For the first time, a screening of the amino acid profile of Gambierdiscus from the Pacific Ocean was conducted, which showed 14 amino acids for all strains except histidine, which was only present in G. cf. caribeaus. We report the presence of Gambierdiscus and Fukuyoa species in the Mexican Pacific, where ciguatera fish poisoning (CFP) cases have occurred. Full article
(This article belongs to the Special Issue Commemorating the Launch of the Section "Marine Toxins")
16 pages, 5383 KiB  
Article
Enhanced Corrosion Resistance of CuAl/BN Coatings through the Addition of Rare Earth Elements and High-Temperature Oxidation Treatment
by Yongjun Liu, Chuanbing Huang, Hao Yang, Xiaoming Sun, Huifeng Zhang, Yonghui Sun, Weigang Zhang, Hao Lan and Shouquan Yu
Crystals 2024, 14(9), 808; https://doi.org/10.3390/cryst14090808 - 12 Sep 2024
Viewed by 318
Abstract
Abradable seal coatings represent a critical technology within the realm of advanced power systems, designed to minimize airflow channel leakage, thereby reducing energy consumption and enhancing overall efficiency. In the present study, CuAl/BN, CuAlLaF3/BN, and CuAlY/BN abradable seal coatings were prepared [...] Read more.
Abradable seal coatings represent a critical technology within the realm of advanced power systems, designed to minimize airflow channel leakage, thereby reducing energy consumption and enhancing overall efficiency. In the present study, CuAl/BN, CuAlLaF3/BN, and CuAlY/BN abradable seal coatings were prepared using plasma spraying technology. Both the as-deposited coatings and high-temperature oxidation-treated coatings were comprehensively investigated by means of scanning electron microscopy (SEM), open-circuit potentials (OCP), potentiodynamic polarization, electrochemical impedance spectroscopy (EIS), salt-spray corrosion testing, and bond strength evaluations. The results show that the addition of rare earth elements to the CuAl/BN coatings does not enhance the corrosion resistance of the coatings and even leads to a decrease in the corrosion resistance of the coatings. In contrast, the CuAlY/BN coatings exhibited a significant improvement in corrosion resistance following an oxidation treatment at 550 °C. This enhancement is attributed to the yttrium (Y) element, which facilitates the formation of passivation films and confers a resistance effect, thereby bolstering the coatings’ resistance to corrosion. The bond strength of the high-temperature oxidation-treated CuAlY/BN coating was improved by about 30% after 960 h of salt-spray corrosion. Full article
(This article belongs to the Section Crystalline Metals and Alloys)
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Figure 1

Figure 1
<p>Cross-section images of the (<b>a</b>) CuAl/BN coating, (<b>b</b>) CuAlY/BN coating, and (<b>c</b>) CuAlLaF<sub>3</sub>/BN coating; (<b>d</b>) X-ray diffraction (XRD) images of the three coatings.</p>
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<p>(<b>a</b>) Open-circuit potential test of the three coatings. (<b>b</b>) Results of the coating polarization curve test.</p>
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<p>Electrochemical impedance spectroscopy (EIS) data for the three coatings: (<b>a</b>) Nyquist plots; (<b>b</b>,<b>c</b>) Bode plots; (<b>d</b>) impedance spectroscopy equivalent circuits.</p>
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<p>(<b>a</b>) Surface scan of CuAl/BN after corrosion for 960 h. (<b>b</b>) Cross-section of CuAlLaF<sub>3</sub>/BN after corrosion for 960 h. (<b>c</b>) Surface scan of CuAlY/BN after corrosion for 960 h.</p>
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<p>XRD patterns of the as-sprayed coatings.</p>
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<p>Bond strength of the as-sprayed coatings versus salt-spray test time.</p>
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<p>(<b>a</b>) Open-circuit potential test of the three coatings. (<b>b</b>) Results of the coating polarization curve test.</p>
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<p>Electrochemical impedance spectroscopy (EIS) data for the three coatings: (<b>a</b>) Nyquist plots; (<b>b</b>,<b>c</b>) Bode plots; (<b>d</b>) impedance spectroscopy equivalent circuits.</p>
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<p>Localized electrochemical impedance spectroscopy (LEIS): (<b>a</b>) CuAl/BN coating; (<b>b</b>) CuAl/BN coating after oxidation; (<b>c</b>) CuAlLaF<sub>3</sub>/BN coating; (<b>d</b>) CuAlLaF<sub>3</sub>/BN coating after oxidation; (<b>e</b>) CuAlY/BN coating; (<b>f</b>) CuAlY/BN coating after oxidation.</p>
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<p>Bond strength of coatings after oxidation pretreatment as a function of salt-spray test time.</p>
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24 pages, 7912 KiB  
Article
Altered Endoplasmic Reticulum Integrity and Organelle Interactions in Living Cells Expressing INF2 Variants
by Quynh Thuy Huong Tran, Naoyuki Kondo, Hiroko Ueda, Yoshiyuki Matsuo and Hiroyasu Tsukaguchi
Int. J. Mol. Sci. 2024, 25(18), 9783; https://doi.org/10.3390/ijms25189783 - 10 Sep 2024
Viewed by 295
Abstract
The cytoskeleton mediates fundamental cellular processes by organizing inter-organelle interactions. Pathogenic variants of inverted formin 2 (INF2) CAAX isoform, an actin assembly factor that is predominantly expressed in the endoplasmic reticulum (ER), are linked to focal segmental glomerulosclerosis (FSGS) and Charcot–Marie–Tooth (CMT) neuropathy. [...] Read more.
The cytoskeleton mediates fundamental cellular processes by organizing inter-organelle interactions. Pathogenic variants of inverted formin 2 (INF2) CAAX isoform, an actin assembly factor that is predominantly expressed in the endoplasmic reticulum (ER), are linked to focal segmental glomerulosclerosis (FSGS) and Charcot–Marie–Tooth (CMT) neuropathy. To investigate how pathogenic INF2 variants alter ER integrity, we used high-resolution live imaging of HeLa cells. Cells expressing wild-type (WT) INF2 showed a predominant tubular ER with perinuclear clustering. Cells expressing INF2 FSGS variants that cause mild and intermediate disease induced more sheet-like ER, a pattern similar to that seen for cells expressing WT-INF2 that were treated with actin and microtubule (MT) inhibitors. Dual CMT-FSGS INF2 variants led to more severe ER dysmorphism, with a diffuse, fragmented ER and coarse INF2 aggregates. Proper organization of both F-actin and MT was needed to modulate the tubule vs. sheet conformation balance, while MT arrays regulated spatial expansion of tubular ER in the cell periphery. Pathogenic INF2 variants also induced mitochondria fragmentation and dysregulated mitochondria distribution. Such mitochondrial abnormalities were more prominent for cells expressing CMT-FSGS compared to those with FSGS variants, indicating that the severity of the dysfunction is linked to the degree of cytoskeletal disorganization. Our observations suggest that pathogenic INF2 variants disrupt ER continuity by altering interactions between the ER and the cytoskeleton that in turn impairs inter-organelle communication, especially at ER–mitochondria contact sites. ER continuity defects may be a common disease mechanism involved in both peripheral neuropathy and glomerulopathy. Full article
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Figure 1
<p>Domain structure and locations of disease variants of human INF2. (<b>A</b>) Human INF2 is a multidomain and homo-dimeric protein that mainly consists of two major components: a regulatory domain that comprises DID and DAD and an actin-organizing unit that includes the FH1 and FH2 domains. The amino acid numbering is shown below each box. There are two isoforms of human INF2: INF2-CAAX (ER-resident) and INF2-non-CAAX (cytoplasmic form) [<a href="#B18-ijms-25-09783" class="html-bibr">18</a>,<a href="#B36-ijms-25-09783" class="html-bibr">36</a>]. INF2-CAAX predominates in the kidney [<a href="#B30-ijms-25-09783" class="html-bibr">30</a>]. INF2 variants exclusively cluster in the DID domain: CMT + FSGS variants are located in the N-terminal half of DID Leu57-Glu183, whereas FSGS variants are found in the C-terminal half of the DID Glu184-Leu245. DID: Diaphanous inhibitory domain. DAD: Diaphanous autoregulatory domain. FH1: Formin homology 1. FH2: Formin homology 2. (<b>B</b>) Schematic diagram showing the domain organization of the ER, with tubular and sheet morphologies. ER membranes are enriched in the perinuclear region (sheet ER), whereas the peripheral ER spreads throughout the cytoplasm as an interconnected tubular network. (<b>C</b>) Subdomains of the peripheral ER. Three components comprising the peripheral ER are shown. A tubule-to-sheet conformation is dynamically regulated by ER proteins, as well as actin-microtubule networks. A cluster of tubules forms the matrix [<a href="#B1-ijms-25-09783" class="html-bibr">1</a>,<a href="#B2-ijms-25-09783" class="html-bibr">2</a>,<a href="#B5-ijms-25-09783" class="html-bibr">5</a>].</p>
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<p>Wild-type INF2 localizes in the peripheral ER in living HeLa cells. (<b>A</b>) Colocalization of wild-type INF2 with the ER compartment. HeLa cells were transiently co-transfected with eGFP tagged, wild-type INF2 (WT, CAAX isoform) (green) and the ER-marker mCherry-calreticulin (red). High-resolution imaging with DragonFly spinning-disk microscopy revealed that WT-INF2 resides in a dispersed reticular network having both tubule and sheet structures (asterisk) that were labeled with the ER marker calreticulin. These structures appear as lace-like, interconnected tubules with a three-way junction (TWJ) and perinuclear sheet. The ER pattern in WT-INF2-expressing HeLa cells is indistinguishable from the control cells expressing calreticulin alone. Bars = 10 µm and 1 µm. (<b>B</b>). Quantitative analysis of colocalization of WT-INF2 with the ER compartment. Colocalization of INF2 with the ER marker (calreticulin) and nuclei (Hoechst) was analyzed using Fiji (version 2.15.1, NIH). The proportion of the merged punctae was calculated using the Manders coefficient. INF2 punctae co-localize preferentially with the ER marker, and the distribution is distinct from the nuclear compartment (<span class="html-italic">p</span> ≤ 0.0001). Data were analyzed by ANOVA test (Prism 8, <span class="html-italic">n</span> = 10 images per subgroup); ns, not significant; ****, <span class="html-italic">p</span> ≤ 0.0001.</p>
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<p>ER morphology in living HeLa cells expressing wild-type and pathogenic INF2 variants. eGFP tagged, wild-type INF2 (WT, CAAX isoform) and pathogenic variants R218W (FSGS), T161N (intermediate), and G73D (CMT + FSGS) (green) were transiently co-transfected with the ER-maker mCherry-calreticulin in HeLa cells. High-resolution live images were captured with a DragonFly microscope. Boxed areas in the peripheral ER are magnified. Cells expressing WT-INF2 show a disperse ER pattern composed of a lace-like, interconnected tubule meshwork. Cells expressing FSGS variants (R218W and T161N) exhibit a predominant sheet-like pattern with fine granular aggregates, while some residual tubular structures remain (asterisk). The clustering of tubules occurs frequently at the cell edges. In contrast, cells expressing the CMT + FSGS variant (G73D) accumulate more sheet-like materials with coarse granular aggregates, which often appear to be fragmented or swollen (double asterisk). Boxes indicate the magnified areas. Bars = 10 µm and 1 µm.</p>
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<p>Comparison of ER phenotypes among live HeLa cells expressing wild-type or variant INF2. (<b>A</b>) Quantification of ER patterns. Representative images of three ER patterns in live HeLa cells expressing WT-INF2 and pathogenic variants are shown. The diffuse reticular ER morphology usually seen in normal control cells is classified as Tubular ER “Class 1”; “Class 2 Mixed tubule and sheet” is defined by a mixed pattern in which tubules coexist with sheets; “Class 3 Sheet” shows predominantly round or ellipsoid-shaped sheet-like materials and aggregation. Boxes present the magnification areas. Cells expressing WT-INF2 or pathogenic variants were categorized by visual inspection, and the proportion of the subtypes was compared. At least 50 cells from at least three experiments were analyzed for each INF2 variant. Statistical analysis was performed using Fisher’s exact test (R, version 4.3.0, 2023); *, <span class="html-italic">p</span> ≤ 0.05; ****, <span class="html-italic">p</span> ≤ 0.0001. (<b>B</b>) Comparison of INF2-ER colocalization among INF2 variants. Intensity correlation of INF2 wild-type and pathogenic variants (eGFP) with an ER marker (mCherry-calreticulin) was analyzed using the Manders coefficient (Fiji, version 2.15.1, NIH). The proportion of INF2 colocalization with the ER was compared between cells expressing WT-INF2 and cells expressing the pathogenic variants. Mild variants R218W and N202S co-localize preferentially with the ER to a similar degree as that for WT-INF2. In contrast, cells expressing the severe G73D variant showed much less ER colocalization (<span class="html-italic">p</span> &lt; 0.05). The T161N variant showed an intermediate colocalization frequency. Data were analyzed by an ANOVA test (Prism 8, <span class="html-italic">n</span> = 10 images/group); ns, not significant; *, <span class="html-italic">p</span> ≤ 0.05. The schematic representation shows hypothetical models for INF2 colocalization with ER markers.</p>
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<p>Effects of the actin inhibitor CytoD on the peripheral ER pattern in living HeLa cells expressing wild-type INF2 variants. Living HeLa cells were transiently transfected with eGFP-INF2 WT (green) and either mScarlet-LifeAct or mCherry-calreticulin (red). The effects of actin polymerization in the ER morphology were examined after treating the cells with cytochalasin D (CytoD; 1 mM for 30 min) [<a href="#B5-ijms-25-09783" class="html-bibr">5</a>]. Before CytoD treatment, cells expressing eGFP-WT-INF2 generated robust, central stress fibers (arrows), as well as peripheral bundles (arrowheads) to produce a disperse, reticular pattern of ER that was labeled with eGFP-INF2. After CytoD treatment, the ER pattern in the WT-INF2 cells acquired a sheet-like appearance (S) rather than tubular structures (T), as labeled with eGFP-INF2. The calreticulin labeling showed a predominant sheet-like pattern with scattered granular aggregates. The data indicate that actin depolymerization disrupts the tubule ER pattern in WT-INF2 expressing cells, leading to a tubule-to-sheet ER transformation. Boxes indicate the magnified areas. Bars = 10 µm and 1 µm.</p>
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<p>Relationship of actin organization and peripheral ER structures in living HeLa cells expressing wild-type or pathogenic INF2 variants. Living HeLa cells were transiently transfected with eGFP-WT-INF2, intermediate (T161N), or severe (G73D) variants (green) and either an F-actin marker (mScarlet-LifeAct) or an ER marker (mCherry-calreticulin, red). T161N variant cells generated fewer central actin cables (arrowheads) and instead have more sheets or granular ER components, in addition to small punctate aggregation of INF2. G73D variant cells produced shorter and thinner F-actin filaments (arrows) and accumulated more coarse granular sheet-like ER components with fragmented INF2 aggregates in the peripheral ER. The predominant sheet-like pattern mirrored that of control WT-INF2 cells treated with CytoD to induce actin depolymerization (<a href="#ijms-25-09783-f005" class="html-fig">Figure 5</a>). Boxes indicate the magnified areas. Bars = 10 µm and 1 µm.</p>
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<p>Effects of a microtubule (MT) inhibitor on the peripheral ER structure in living HeLa cells expressing wild-type and pathogenic INF2 variants. HeLa cells were transiently cotransfected with eGFP-INF2 WT (green) and mScarlet EMTB (red). Cells expressing eGFP-WT-INF2 showed a reticular, tubular ER pattern, along with an MT array having appropriate spacing. The effects of MT depolymerization on ER morphology were examined by treating cells with nocodazole (Noc, 2.5 µg/mL) for 30 min. In eGFP-WT-INF2 cells, INF2-labeled ER structures form an expansive network reaching to the farthest edge of the cells. Noc-induced MT depolymerization altered the tubular structure (T) such that it was sparser in some areas. Sheet (S) or matrix structures formed in the remaining network. There was partial retraction of the ER network towards the cell center (asterisk), suggesting a role for MT in enabling the ER to expand throughout the cell. Boxes indicate the magnified areas. Dashed lines depict the cell contour. Bars = 10 µm and 2 µm.</p>
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<p>Effects of a microtubule inhibitor on the peripheral ER in living HeLa cells expressing wild-type or pathogenic INF2 variants. HeLa cells were transiently cotransfected with eGFP-INF2 (WT, T161N, G73D) (green) and mCherry-calreticulin (red). The effects of inhibiting MT polymerization on ER morphology were examined by treating cells with nocodazole (Noc, 2.5 µg/mL) for 30 min. In eGFP-WT-INF2 cells, Noc decreased peripheral tubular components with concomitant mild and focal enrichment of sheet or matrix structures, while leaving tubule structures intact in some areas (asterisk). T161N cells with Noc treatment show more tiny granular sheet-like ER components (mCherry-calreticulin) and diffusely disperse INF2 distribution (eGFP). G73D cells accumulated more coarse, sheet-like ER materials and punctate INF2 aggregates throughout the cytoplasm than T161N cells. Boxes indicate the magnified areas. Bars = 10 µm and 1 µm.</p>
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<p>ER–mitochondria interaction in living HeLa cells expressing WT-INF2 or pathogenic variants. (<b>A</b>). ER and mitochondria colocalization. ER and mitochondria were labeled by eGFP-INF2 and MitoTracker, respectively. Cells expressing WT-INF2 had lace-like polygonal ER tubules spread towards the periphery, with mitochondria aligned in the perinuclear region, forming ample ERMCSs (ER–mitochondria contact sites). Cells expressing FSGS variants (R218W, N202S, T161N) exhibited a predominant aberrant mitochondria cluster (C) at the periphery, with fragmentation and disrupted ERMCSs. T161N variant cells showed more pronounced misdistribution and fragmentation than WT. CMT + FSGS variant (G73D) cells showed even more marked dysmorphic ER changes, with a diffuse, coarse granular ER appearance, leading to severe dissociation (D) of ERMCS. Bars = 10 µm and 2 µm. (<b>B</b>). Three subclasses of mitochondrial shape were observed: Class 1: predominant tubular-shape, Class 2: mixture of tubular and fragmented mitochondria, and Class 3: fragmented. Mitochondria were more fragmented in cells expressing INF2 variants (R218W, N202S, T161N, G73D) than those expressing WT-INF2. (<b>C</b>) Mitochondrial distribution is subclassified as either perinuclear (normal distribution) or peripheral (misdistribution). Data are from three independent experiments (<span class="html-italic">n</span> ≥ 30 cells per each variant); ns, not significant; *, <span class="html-italic">p</span> ≤ 0.05.</p>
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<p>Impaired mitochondrial respiratory function in HeLa cells expressing INF2 variants. Living HeLa cells were co-transfected with eGFP-tagged WT-INF2, T161N, or G73D variants. After 12 h, cells were analyzed for mitochondrial function by measuring the OCR in response to the indicated reagents. (<b>A</b>) Diagram of mitochondrial respiration. Cells were treated consecutively with a series of complex inhibitor or coupling reagent (oligomycin, carbonyl cyanide 4-(trifluoromethoxy) phenylhydrazone (FCCP), and rotenone + antimycin A) and measured with a Flux analyzer (XFp Agilent). Data are normalized by transfection efficiency, and the mean ± SE from three independent experiments is shown. (<b>B</b>) Comparison of mitochondrial respiration for INF2 variants. WT-INF2 cells exhibited high basal respiration and respiratory capacity during the treatments, while G73D variant cells showed much lower respiratory parameters, suggesting more severely compromised mitochondrial function (<span class="html-italic">p</span> ≤ 0.05). Cells expressing the T161N variant had an intermediate performance. Data were analyzed by ANOVA test (Prism 8, <span class="html-italic">n</span> = 3 experiments); ns, not significant; *, <span class="html-italic">p</span> ≤ 0.05.</p>
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<p>Effects of INF2 variants on ER morphology and organelle contacts. (<b>A</b>) ER integrity and ER–organelle interactions. Under physiological conditions, the ER is canonically classified as two simple structures: tubule (peripheral ER) and sheet (perinuclear cisterna ER). Tubular ER exhibits a reticular pattern with polygons connected by a three-way junction (TWJ). The ER network is mainly shaped and maintained by interplay with actin and MT. The ER networks contact organelles in close opposition and contribute to a variety of processes, including (1) mitochondria dynamics, (2) vesicle trafficking (with arrow shows the movement direction), and (3) plasma membrane Ca exchange. (<b>B</b>) Schematic of representative ER pattern in cells expressing INF2 variants. WT-INF2 cells showed a disperse reticular network pattern, appearing as lace-like interconnected tubules with a TWJ and occasional sheet. FSGS variant cells have strikingly altered ER patterns, evidenced by more abundant sheet-like structures with polygonal structures left intact in some areas. Cells expressing CMT + FSGS variants show a diffuse dysmorphic ER appearance, with a loss of continuity marked by irregular dilatation and fragmentation.</p>
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16 pages, 2191 KiB  
Article
Heat-Killed Lactobacillus acidophilus Promotes Growth by Modulating the Gut Microbiota Composition and Fecal Metabolites of Piglets
by Huabiao Miao, Jing Liang, Ganqiu Lan, Qian Wu and Zunxi Huang
Animals 2024, 14(17), 2528; https://doi.org/10.3390/ani14172528 - 30 Aug 2024
Viewed by 431
Abstract
Probiotics can improve animal growth performance and intestinal health. However, understanding the effects of paraprobiotics on the growth performance and gut microbiota of piglets and how the paraprobiotics exert their impact are still limited. The present study was conducted to investigate the effects [...] Read more.
Probiotics can improve animal growth performance and intestinal health. However, understanding the effects of paraprobiotics on the growth performance and gut microbiota of piglets and how the paraprobiotics exert their impact are still limited. The present study was conducted to investigate the effects of heat-killed Lactobacillus acidophilus IFFI 6005 supplementation on the growth performance, intestinal microbiota, and fecal metabolites of piglets. First, a feed-additive sample of heat-killed Lactobacillus acidophilus IFFI 6005 was prepared by culture. Second, 96 (initial BW = 14.38 ± 0.67 kg, weaning age of 40 days) healthy piglets were selected and randomized into four treatment groups. Each treatment group consisted of three replicates (n = 8). Pigs were fed a basal diet (NC), basal diet plus antibiotics (PC), basal diet plus Lactobacillus acidophilus IFFI 6005 at 600 g/t (LA, 1.0 × 1010 cfu/g), and basal diet plus heat-killed Lactobacillus acidophilus IFFI 6005 at 600 g/t (HKLA), respectively; the trial lasted for 30 days. The results showed that the ratios of feed to gain (F:G) and diarrhea rate of both the HKLA and PC groups were significantly lower compared with the NC and LA groups (p < 0.05); however, there was no significant difference between the HKLA and PC group (p > 0.05). In addition, the average daily weight gain (ADG) of the HKLA group was significantly higher (p < 0.05) than that of the other three groups in terms of growth performance. Finally, 16S rRNA sequencing and metabolome analysis based on fecal samples further elaborated that the addition of heat-killed Lactobacillus acidophilus IFFI 6005 to the feed improved the intestinal microbial diversity and abundance (p < 0.05) and reduced the abundance of pathogenic bacteria (p < 0.05), but it did not affect the abundance of Lactobacillus (p > 0.05). Through the comparison of microbial abundance and metabolite content between the two groups (NC_vs_HKLA), the largest differences were found in six microorganisms and 10 metabolites in the intestine (p < 0.05). These differential metabolites were involved in the digestion, absorption and utilization of protein and starch, as well as in oxidative stress. In summary, addition of heat-killed Lactobacillus acidophilus IFFI 6005 as a new feed additive in piglets has beneficial effects on the growth performance, intestinal bacteria and metabolites, and can be used as an alternative to antibiotics. Full article
(This article belongs to the Section Pigs)
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<p>Culture and preparation of heat-killed <span class="html-italic">Lactobacillus acidophilus</span>. (<b>a</b>) High-density fermentation results of <span class="html-italic">Lactobacillus acidophilus</span> IFFI 6005 in a 50-L fermenter. To determine the biomass and pH, culture samples were collected every 4 h from 0 to 48 h. Biomass and pH are represented by squares and triangles, respectively. (<b>b</b>) Antibacterial activity of the fermentation concentrate with heat-killed <span class="html-italic">Lactobacillus acidophilus</span> IFFI 6005. The test method for antibacterial activity was the agar well diffusion method. CK represents the diameter of the inhibition zone measured by adding an equal amount of blank medium, indicated by a red histogram. The results represent the mean ± SD.</p>
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<p>Alpha diversity in intestinal microorganisms from each treatment group (<span class="html-italic">n</span> = 6 in each treatment). (<b>a</b>) Sob index; (<b>b</b>) PD-tree index; (<b>c</b>) Chao1 index; abbreviations: NC = control group; HKLA = test group. * and ** represent <span class="html-italic">p</span> &lt; 0.05 and <span class="html-italic">p</span> &lt; 0.01, respectively, using a <span class="html-italic">t</span>-test.</p>
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<p>Differential analysis of the microbiota in weaned piglet fecal samples (<span class="html-italic">n</span> = 6 in each treatment). (<b>a</b>) PCoA of the gut bacterial communities between two groups; (<b>b</b>) Venn analysis at the OTU level; (<b>c</b>) analysis of species composition of differential operational taxonomic units at the genus level; (<b>d</b>) statistical analysis of differential species of abundance; (<b>e</b>) the prediction of the function for specific OTUs in the Greengene database. (<b>f</b>) Comparison of the contents of the top 5 <span class="html-italic">Lactobacillus</span> in the two groups. Abbreviations: NC = control group; HKLA = test group. PCoA = intergroup principal coordinate analysis; PCo1 = the principal coordinate components that explain the largest possible variation in the data; PCo2 = the principal coordinate components that explain the largest proportion of the remaining variability. The mean (SD) values of each group were determined using a Student’s <span class="html-italic">t</span>-test. Median (IQR) values were determined using a Mann–Whitney U test. *, **, and *** represent <span class="html-italic">p</span> &lt; 0.05, <span class="html-italic">p</span> &lt; 0.01, and <span class="html-italic">p</span> &lt; 0.001, respectively, using <span class="html-italic">t</span>-tests.</p>
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<p>Metabolites in weaned piglet fecal samples (<span class="html-italic">n</span> =6 in each treatment). (<b>a</b>) Score plot of the PLS-DA model; (<b>b</b>) histogram of differential metabolites with <span class="html-italic">p</span> &lt; 0.05 and |log<sub>2</sub>(FC)| &gt; 2 between two groups; (<b>c</b>) enrichment of differential metabolites in metabolic pathways; (<b>d</b>) The key differential metabolite load map; (<b>e</b>) top 10 metabolite and bacterial boxplots showed differences between two groups; abbreviations: NC = control group; HKLA = test group. The mean (SD) values of each group were determined using a Student’s <span class="html-italic">t</span>-test. Median (IQR) values were determined using a Mann–Whitney U test.</p>
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<p>Association analysis of differential microorganisms and metabolites (<span class="html-italic">n</span> = 6 in each treatment). (<b>a</b>) Correlation analysis of key fecal differential microorganisms with metabolites; (<b>b</b>) correlation analysis of the differential metabolite chenodeoxycholate with the top 10 differential microorganisms. The legend shows the magnitude of Spearman’s correlation coefficient; red indicates a positive correlation, and blue indicates a negative correlation. *, **, and *** represent <span class="html-italic">p</span> &lt; 0.05, <span class="html-italic">p</span> &lt; 0.01, and <span class="html-italic">p</span> &lt; 0.001, respectively, using <span class="html-italic">t</span>-tests.</p>
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13 pages, 3706 KiB  
Article
Anti-Warburg Mechanism of Ginsenoside F2 in Human Cervical Cancer Cells via Activation of miR193a-5p and Inhibition of β-Catenin/c-Myc/Hexokinase 2 Signaling Axis
by Nari Shin, Hyo-Jung Lee, Deok Yong Sim, Chi-Hoon Ahn, Su-Yeon Park, Wonil Koh, Jaeho Khil, Bum-Sang Shim, Bonglee Kim and Sung-Hoon Kim
Int. J. Mol. Sci. 2024, 25(17), 9418; https://doi.org/10.3390/ijms25179418 - 30 Aug 2024
Viewed by 309
Abstract
Though Ginsenoside F2 (GF2), a protopanaxadiol saponin from Panax ginseng, is known to have an anticancer effect, its underlying mechanism still remains unclear. In our model, the anti-glycolytic mechanism of GF2 was investigated in human cervical cancer cells in association with miR193a-5p and [...] Read more.
Though Ginsenoside F2 (GF2), a protopanaxadiol saponin from Panax ginseng, is known to have an anticancer effect, its underlying mechanism still remains unclear. In our model, the anti-glycolytic mechanism of GF2 was investigated in human cervical cancer cells in association with miR193a-5p and the β-catenin/c-Myc/Hexokinase 2 (HK2) signaling axis. Here, GF2 exerted significant cytotoxicity and antiproliferation activity, increased sub-G1, and attenuated the expression of pro-Poly (ADPribose) polymerase (pro-PARP) and pro-cysteine aspartyl-specific protease (procaspase3) in HeLa and SiHa cells. Consistently, GF2 attenuated the expression of Wnt, β-catenin, and c-Myc and their downstream target genes such as HK2, pyruvate kinase isozymes M2 (PKM2), and lactate dehydrogenase A (LDHA), along with a decreased production of glucose and lactate in HeLa and SiHa cells. Moreover, GF2 suppressed β-catenin and c-Myc stability in the presence and absence of cycloheximide in HeLa cells, respectively. Additionally, the depletion of β-catenin reduced the expression of c-Myc and HK2 in HeLa cells, while pyruvate treatment reversed the ability of GF2 to inhibit β-catenin, c-Myc, and PKM2 in GF2-treated HeLa cells. Notably, GF2 upregulated the expression of microRNA139a-5p (miR139a-5p) in HeLa cells. Consistently, the miR139a-5p mimic enhanced the suppression of β-catenin, c-Myc, and HK2, while the miR193a-5p inhibitor reversed the ability of GF2 to attenuate the expression of β-catenin, c-Myc, and HK2 in HeLa cells. Overall, these findings suggest that GF2 induces apoptosis via the activation of miR193a-5p and the inhibition of β-catenin/c-Myc/HK signaling in cervical cancer cells. Full article
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<p>Effect of GF2 on cytotoxicity and colony formation in HeLa and SiHa cells. (<b>A</b>) Chemical structure of GF2 (MW = 785.03). (<b>B</b>) Cytotoxic effect of GF2 in HeLa and SiHa cells. The cells were exposed to various concentrations of GF2 for 24 h, and cell viability was assessed by MTT assay. * <span class="html-italic">p</span> &lt; 0.05, ** <span class="html-italic">p</span> &lt; 0.01, and *** <span class="html-italic">p</span> &lt; 0.001 versus the untreated control. (<b>C</b>) Antiproliferative effect of GF2 in HeLa and SiHa cells. HeLa and SiHa cells were treated with GF2 (0, 50, and 70 μM), and then colony formation took place over 1 week. The colonies were visualized by staining with Diff-Quick solution (Sysmex, Kobe, Japan). The data represent the means ± SD.</p>
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<p>Effect of GF2 on apoptosis in HeLa and SiHa cells. (<b>A</b>) Effect of GF2 on PARP and caspase 3 in HeLa and SiHa cells. (<b>B</b>) Effect of GF2 on sub-G1 phase in HeLa and SiHa cells.</p>
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<p>Effect of GF2 on band level of Wnt, β-catenin, c-Myc, HK2, PKM2, and LDHA in HeLa and SiHa cells. (<b>A</b>) Effect of GF2 on Wnt, β-catenin, and c-Myc in HeLa and SiHa cells. (<b>B</b>) Effect of GF2 on HK2, PKM2, and LDHA in HeLa and SiHa cells. (<b>C</b>) Effect of GF2 on glucose production in HeLa and SiHa cells using a colorimetric assay. The results represent the ±SD; *** <span class="html-italic">p</span> &lt; 0.001. (<b>D</b>) Effect of GF2 on lactate production in HeLa and SiHa cells using a colorimetric assay. The results represent mean ± SD; ** <span class="html-italic">p</span> &lt; 0.01, *** <span class="html-italic">p</span> &lt; 0.001.</p>
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<p>Effect of GF2 on the stability of β-catenin, c-Myc, and HK2 in HeLa cells.</p>
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<p>Binding index and correlation efficient between β-catenin and c-Myc and the effect of β-catenin siRNA or pyruvate on glycolysis-related proteins in HeLa cells. (<b>A</b>) RNA-seq data (cBioportal) confirm a strong correlation between β-catenin and c-Myc with a Spearman correlation coefficient of 0.09. (<b>B</b>) Binding between β-catenin and c-Myc in HeLa cells. Immunoprecipitation was performed in HeLa cells treated with GF2 for 24 h and then subjected to Western blotting to detect β-catenin and c-Myc concentrations in whole-cell lysates. (<b>C</b>) Effect of β-catenin depletion on β-catenin, c-Myc, and HK2 in GF2-treated HeLa cells. (<b>D</b>) Effect of pyruvate treatment on β-catenin and PKM2 in GF2-treated HeLa cells.</p>
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<p>The important role of miR-139a-5p in GF2-induced apoptosis in HeLa cells. (<b>A</b>) Direct binding of miR-139a-5p to the 3′-Untranslated region of β-catenin. (<b>B</b>) Endogenous expression level of miR139a-5p in HeLa cells. The expression level of miR139a-5p was measured in intact HeLa cells by RT-PCR, *** <span class="html-italic">p</span> &lt; 0.001. (<b>C</b>) Effect of miR139a-5p mimic on β-catenin, HK2, and c-Myc in HeLa cells. The expression of β-catenin, c-Myc, HK2, and pro-PARP was evaluated in HeLa cells with or without the miR139a-5p mimic by Western blotting. (<b>D</b>) Effect of miR139a-5p inhibitor on β-catenin, c-Myc, HK2, and pro-PARP in GF2-treated HeLa cells. The expression of β-catenin, c-Myc, HK2, and pro-PARP was evaluated in HeLa cells with or without GF2 by Western blotting.</p>
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<p>Schematic diagram on the anti-Warburg effect of GF2 via the activation of miR-139a-5p and the suppression of the β-catenin/c-Myc/HK2 signaling axis.</p>
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17 pages, 921 KiB  
Article
Characterisation of the Atmosphere in Very High Energy Gamma-Astronomy for Imaging Atmospheric Cherenkov Telescopes
by Dijana Dominis Prester, Jan Ebr, Markus Gaug, Alexander Hahn, Ana Babić, Jiří Eliášek, Petr Janeček, Sergey Karpov, Marta Kolarek, Marina Manganaro and Razmik Mirzoyan
Universe 2024, 10(9), 349; https://doi.org/10.3390/universe10090349 - 30 Aug 2024
Viewed by 485
Abstract
Ground-based observations of Very High Energy (VHE) gamma rays from extreme astrophysical sources are significantly influenced by atmospheric conditions. This is due to the atmosphere being an integral part of the detector when utilizing Imaging Atmospheric Cherenkov Telescopes (IACTs). Clouds and dust particles [...] Read more.
Ground-based observations of Very High Energy (VHE) gamma rays from extreme astrophysical sources are significantly influenced by atmospheric conditions. This is due to the atmosphere being an integral part of the detector when utilizing Imaging Atmospheric Cherenkov Telescopes (IACTs). Clouds and dust particles diminish atmospheric transmission of Cherenkov light, thereby impacting the reconstruction of the air showers and consequently the reconstructed gamma-ray spectra. Precise measurements of atmospheric transmission above Cherenkov observatories play a pivotal role in the accuracy of the analysed data, among which the corrections of the reconstructed energies and fluxes of incoming gamma rays, and in establishing observation strategies for different types of gamma-ray emitting sources. The Major Atmospheric Gamma Imaging Cherenkov (MAGIC) telescopes and the Cherenkov Telescope Array Observatory (CTAO), both located on the Observatorio del Roque de los Muchachos (ORM), La Palma, Canary Islands, use different sets of auxiliary instruments for real-time characterisation of the atmosphere. In this paper, historical data taken by MAGIC LIDAR (LIght Detection And Ranging) and CTAO FRAM (F/Photometric Robotic Telescope) are presented. From the atmospheric aerosol transmission profiles measured by the MAGIC LIDAR and CTAO FRAM aerosol optical depth maps, we obtain the characterisation of the clouds above the ORM at La Palma needed for data correction and optimal observation scheduling. Full article
(This article belongs to the Collection Women Physicists in Astrophysics, Cosmology and Particle Physics)
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<p>Overview plot used in data quality checks for a single observing night at the MAGIC telescope site. Four panels (enumerated from top to bottom) allow for an overview of the most important parameters, which describe the data-taking conditions and/or impact on the data quality. Panel 1: Zenith angle of the telescope pointing, with vertical lines marking the wobble position or source changes. Panel 2: Mean direct current (DC; green and red) and the median discriminator threshold (TH; blue) for the camera PMTs. The colour of the DC plot roughly captures the dark time (green) and moon time (orange) observations, with dark time observations further separated into extragalactic (dark green) and galactic (light green). Panel 3: Coincidence trigger rates between the two MAGIC telescopes. Panel 4: Atmospheric aerosol transmission, based on measurements by the LIDAR and the pyrometer. Two transmission measures are given based on the LIDAR profiles, from 3 km and from 9 km above the site, quantifying the dust intrusions (calima) and cloud effects, respectively. Colour shading gives a rough guidance whether the data may be used without corrections, with nominal or increased energy threshold (green, light yellow), may be corrected for atmosphere effects (dark yellow), or cannot be used at all due to poor quality (red).</p>
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<p>Scheme for the determination of new Stable Time Intervals (STIs) and new Monte Carlo (MC) simulated instrument response functions (IRFs): on the top, two possible aerosol extinction profiles at two wavelengths obtained from LIDAR are sketched, with nested VAOD maps obtained from FRAM during a typical CTAO observation block of 25 min. In this particular case, a typical aerosol ground layer of a clear night that ranges up to 2.5 km above ground evolves slowly towards a higher Ångström exponent (smaller average particulate sizes) at a similar vertical profile, whereas a cloud at a typical height of about 10 km moves into the FRAM field of view. The joint analysis of FRAM and LIDAR (and possibly CTC) data divides the tessellated VAOD maps from FRAM first into a slow and a fast component and then produces the aerosol transmission hypercubes through interpolation, both in time and in altitude. The offline science data analysis calculates systematic errors introduced by the not perfectly matching aerosol conditions and the aerosol extinction cube used for the production of the given IRFs, and decides to start new STIs as the systematic error exceeds a predefined limit. Each STI is then analysed with its (simplified) AEC used for the production of a tailored IRF. See text for further details.</p>
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<p>Vertical standard deviation of clouds (“extension”) as a function of mean altitude. A smoothing on 3 × 3 consecutive cells (“k3a”) was applied. Left: summer clouds (July/August); right: clouds from the rest of the year.</p>
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<p>Distributions of normalised extinction vs. vertical distance from mean altitude (see text for definitions). Left: all extinction profiles (colour map), with the binwise means (black), medians (white), and 10% and 90% quantiles (orange). All parameters were fitted to Equation (<a href="#FD1-universe-10-00349" class="html-disp-formula">1</a>). Right: the mean extinction profiles for different cloud cases: low clouds (<math display="inline"><semantics> <mrow> <mover> <mi>H</mi> <mo>¯</mo> </mover> <mo>&lt;</mo> <mn>7</mn> <mspace width="3.33333pt"/> <mi>km</mi> <mspace width="3.33333pt"/> <mi mathvariant="normal">a</mi> <mo>.</mo> <mi mathvariant="normal">s</mi> <mo>.</mo> <mi mathvariant="normal">l</mi> <mo>.</mo> </mrow> </semantics></math>, black), medium altitude clouds (<math display="inline"><semantics> <mrow> <mn>7</mn> <mspace width="3.33333pt"/> <mi>km</mi> <mspace width="3.33333pt"/> <mi mathvariant="normal">a</mi> <mo>.</mo> <mi mathvariant="normal">s</mi> <mo>.</mo> <mi mathvariant="normal">l</mi> <mo>.</mo> <mo>&lt;</mo> <mover> <mi>H</mi> <mo>¯</mo> </mover> <mo>&lt;</mo> <mn>14</mn> <mspace width="3.33333pt"/> <mi>km</mi> <mspace width="3.33333pt"/> <mi mathvariant="normal">a</mi> <mo>.</mo> <mi mathvariant="normal">s</mi> <mo>.</mo> <mi mathvariant="normal">l</mi> <mo>.</mo> </mrow> </semantics></math>) for three different VOD ranges: <math display="inline"><semantics> <mrow> <mi>VOD</mi> <mo>&lt;</mo> <mn>0.2</mn> </mrow> </semantics></math> (lilac), <math display="inline"><semantics> <mrow> <mn>0.2</mn> <mo>&lt;</mo> <mi>VOD</mi> <mo>&lt;</mo> <mn>0.5</mn> </mrow> </semantics></math> (green) and <math display="inline"><semantics> <mrow> <mi>VOD</mi> <mo>&gt;</mo> <mn>0.5</mn> </mrow> </semantics></math> (pink), and summer clouds (orange). These profiles are also shown together with their fits to Equation (<a href="#FD1-universe-10-00349" class="html-disp-formula">1</a>).</p>
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<p>Time series of zero points from scans. The green line connects values accepted during real-time processing; the red line connects values excluding outliers.</p>
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<p>Distribution of VAOD values measured in tiles with different cuts applied.</p>
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<p>Distribution of VAOD values measured in tiles with different cuts applied, normalised to the number of entries after each cut.</p>
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<p>Distribution of VAOD values measured in tiles after all cuts compared with the distribution of the precise VAOD measurements from scans without and with a smearing with a Gaussian with <math display="inline"><semantics> <mrow> <mi>σ</mi> <mo>=</mo> <mn>0.04</mn> </mrow> </semantics></math>.</p>
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<p>Distribution of VAOD values measured in entire images with different cuts applied, normalised to the number of entire images after each cut.</p>
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<p>Distribution of VAOD values measured in entire images after all cuts compared with the distribution of the precise VAOD measurements from scans without and with a smearing with a Gaussian with <math display="inline"><semantics> <mrow> <mi>σ</mi> <mo>=</mo> <mn>0.01</mn> </mrow> </semantics></math>.</p>
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9 pages, 878 KiB  
Article
An Expected Goals on Target (xGOT) Metric as a New Metric for Analyzing Elite Soccer Player Performance
by Anselmo Ruiz-de-Alarcón-Quintero and Blanca De-la-Cruz-Torres
Data 2024, 9(9), 102; https://doi.org/10.3390/data9090102 - 28 Aug 2024
Viewed by 646
Abstract
Introduction. Football analysis is an applied research area that has seen a huge upsurge in recent years. More complex analysis to understand the soccer players’ or teams’ performances during matches is required. The objective of this study was to prove the usefulness of [...] Read more.
Introduction. Football analysis is an applied research area that has seen a huge upsurge in recent years. More complex analysis to understand the soccer players’ or teams’ performances during matches is required. The objective of this study was to prove the usefulness of the expected goals on target (xGOT) metric, as a good indicator of a soccer team’s performance in professional Spanish football leagues, both in the women’s and men’s categories. Method. The data for the Spanish teams were collected from the statistical website Football Reference. The 2023/24 season was analyzed for Spanish leagues, both in the women’s and men’s categories (LigaF and LaLiga, respectively). For all teams, the following variables were calculated: goals, possession value (PV), expected goals (xG) and xGOT. All data obtained for each variable were normalized by match (90 min). A descriptive and correlational statistical analysis was carried out. Results. In the men’s league, this study found a high correlation between goals per match and xGOT (R2 = 0.9248) while in the women’s league, there was a high correlation between goals per match (R2 = 0.9820) and xG and between goals per match and xGOT (R2 = 0.9574). Conclusions. In the LaLiga, the xGOT was the best metric that represented the match result while in the LigaF, the xG and the xGOT were the best metrics that represented the match score. Full article
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<p>R<sup>2</sup> correlation of total goals with possession value (PV), expected goals (xG) and expected goals on target (xGOT) in LigaF (<b>A</b>–<b>C</b>) and LaLiga (<b>D</b>–<b>F</b>).</p>
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<p>Chain on goals model in football.</p>
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13 pages, 2940 KiB  
Project Report
Correlation between Epsilon Wave and Late Potentials in Arrhythmogenic Right Ventricular Cardiomyopathy—Do Late Potentials Define the Epsilon Wave?
by Urszula Skrzypczyńska-Banasik, Olgierd Woźniak, Ilona Kowalik, Aneta Fronczak-Jakubczyk, Karolina Borowiec, Piotr Hoffman and Elżbieta Katarzyna Biernacka
J. Clin. Med. 2024, 13(17), 5038; https://doi.org/10.3390/jcm13175038 - 25 Aug 2024
Viewed by 500
Abstract
Introduction: Arrhythmogenic right ventricular cardiomyopathy (ARVC) is a genetic disorder characterised by progressive fibrosis predominantly of the right ventricular (RV) myocardium, resulting in life-threatening arrhythmias and heart failure. The diagnosis is challenging due to a wide spectrum of clinical symptoms. The important [...] Read more.
Introduction: Arrhythmogenic right ventricular cardiomyopathy (ARVC) is a genetic disorder characterised by progressive fibrosis predominantly of the right ventricular (RV) myocardium, resulting in life-threatening arrhythmias and heart failure. The diagnosis is challenging due to a wide spectrum of clinical symptoms. The important role of ECG was covered in the current diagnostic criteria. The role of the epsilon wave (EW) is still under discussion. Aim: The aim of the study was to examine a potential association between the EW and late ventricular potentials (LPs) in ARVC patients (pts). The correlation between RV dilatation or dysfunction and LPs/EW was also analysed. Methods: The ARVC group consisted of 81 pts (53 men, aged 20–78 years) fulfilling 2010 International Task Force Criteria. 12-lead ECG, LPs, Holter, and ECHO were performed in all pts. The presence of EW was analysed in ECG by 3 investigators. LPs were detected by signal-averaged ECG (SAECG). SAECG was considered positive for LPs when at least two of the three following criteria were met: (1) the filtered QRS duration (fQRS) ≥ 114 msec; (2) the duration of the final QRS fragment in which low-amplitude signals lower than 40 μV are recorded (LAS-40 > 38 msec); and (3) the root mean square amplitude of the last 40 milliseconds of the fQRS complex (RMS-40 < 20 μV). The results were compared with a reference group consisting of 53 patients with RV damage in the course of atrial septum defect (ASD) or Ebstein’s Anomaly (EA). Results: In the ARVC group, a significant relationship was observed between the occurrence of EW and the presence of LPs. EW was more common in the LP+ than in the LP- patients (48.1% vs. 6.9%, p < 0001; OR 12.5; 95% CI [2.691–58.063]). In ARVC pts, RVOT > 36 mm, RVIT > 41 mm, and RV S’ < 9 cm/s were observed significantly more often in the LPs+ than in the LPs− group (OR [95% CI]: 8.3 [2.9–1.5], 6.4 [2.2–19.0] and 3.6 [1.1–12.2], respectively). In the ARVC group, any of fQRS > 114 ms, LAS > 38 ms, and RMS < 20 μV were significantly more frequent in EW+ pts. In multivariate analysis, the independent factors of the EW were LAS-40 and RV S’. In the LPs− subgroup, RVOT > 36 mm was more frequent in ASD/EA than in ARVC (70.4% vs. 25%, p = 0.002). Similarly, in the LPs− subgroup, RVIT > 41 mm was encountered more frequently in ASD/EA than in ARVC (85.2% vs. 48.3%, p = 0.004). Conclusions: In ARVC, there is an association between EW and LPs, with both probably resulting from the same process of fibrofatty substitution of the RV myocardium. Although RV dilatation is common in ASD and EA, it does not correlate with LPs. Full article
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<p>Registration of (<b>A</b>) standard 12-lead ECG and (<b>B</b>) SAECG in an ARVC (EW+, LP+) patient. The arrows indicate epsilon waves (<b>A</b>) and late potentials (<b>B</b>).</p>
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<p>Prevalence of EW and its association with LPs in ARVC and ASD/EA.</p>
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<p>Prevalence of RVOT &gt; 36 mm and association between RVOT diameter and LPs in ARVC and ASD/EA (homogeneity of OR: <span class="html-italic">p</span> = 0.049).</p>
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<p>Prevalence of RVIT &gt; 41 mm and association between RVIT diameter and LPs in ARVC and ASD/EA (homogeneity of OR: <span class="html-italic">p</span> = 0.015).</p>
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<p>Prevalence of RV S’ &lt; 9 cm/s and association between RV S’ and LPs in ARVC and ASD/EA groups (homogeneity of OR: <span class="html-italic">p</span> = 0.049).</p>
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<p>Multivariable logistic regression results for the occurrence of EW.</p>
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<p>Comparison of ROC curves diagnosing the usefulness of explanatory variables in the model defining the occurrence of EW (dependent variable).</p>
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12 pages, 6711 KiB  
Article
Crystal Structure and Microwave Dielectric Characteristics of Novel Ba(Eu1/5Sm1/5Nd1/5Pr1/5La1/5)2Ti4O12 High-Entropy Ceramic
by Qing Wan, Zeping Li, Huifeng Wang, Gang Xiong and Geng Wang
Crystals 2024, 14(9), 754; https://doi.org/10.3390/cryst14090754 - 25 Aug 2024
Viewed by 419
Abstract
High-permittivity Ba(Eu1/5Sm1/5Nd1/5Pr1/5La1/5)2Ti4O12 (BESNPLT) high-entropy ceramics (HECs) were synthesized via a solid-state route. The microstructure, sintering behavior, phase structure, vibration modes, and microwave dielectric characteristics of the BESNPLT HECs [...] Read more.
High-permittivity Ba(Eu1/5Sm1/5Nd1/5Pr1/5La1/5)2Ti4O12 (BESNPLT) high-entropy ceramics (HECs) were synthesized via a solid-state route. The microstructure, sintering behavior, phase structure, vibration modes, and microwave dielectric characteristics of the BESNPLT HECs were thoroughly investigated. The phase structure of the BESNPLT HECs was confirmed to be a single-phase orthorhombic tungsten-bronze-type structure of Pnma space group. Permittivity (εr) was primarily influenced by polarizability and relative density. The quality factor (Q×f) exhibited a significant correlation with packing fraction, whereas the temperature coefficient (TCF) of the BESNPLT HECs closely depended on the tolerance factor and bond valence of B-site. The BESNPLT HECs sintered at 1400 °C, demonstrating high relative density (>97%) and optimum microwave dielectric characteristics with TCF = +38.9 ppm/°C, Q×f = 8069 GHz (@6.1 GHz), and εr = 87.26. This study indicates that high-entropy strategy was an efficient route in modifying the dielectric characteristics of tungsten-bronze-type microwave ceramics. Full article
(This article belongs to the Special Issue Crystal Structure and Dielectric Properties of Ceramics)
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<p>XRD patterns of BESNPLT HECs sintered at 1350–1500 °C.</p>
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<p>(<b>a</b>) Rietveld refinement plots of BESNPLT HECs. (<b>b</b>) Structural diagram of BESNPLT HECs.</p>
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<p>Raman spectra of BESNPLT HECs sintered at varying temperatures.</p>
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<p><span class="html-italic">ρ<sub>re</sub></span> and <span class="html-italic">ρ<sub>bu</sub></span> of BESNPLT HECs sintered at 1350–1500 °C.</p>
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<p>SEM morphology of BESNPLT HECs sintered at (<b>a</b>) 1350 °C; (<b>b</b>) 1400 °C; (<b>c</b>) 1450 °C; (<b>d</b>) 1500 °C.</p>
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<p>EDS mapping of BESNPLT HECs.</p>
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<p>(<b>a</b>) <span class="html-italic">ε<sub>r</sub></span> and <span class="html-italic">ρ<sub>re</sub></span> of BESNPLT HECs; (<b>b</b>) <span class="html-italic">ε<sub>cor</sub></span> and <span class="html-italic">α<sub>the</sub>/<span class="html-italic">V<sub>m</sub></span></span> of BESNPLT HECs.</p>
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<p>(<b>a</b>) <span class="html-italic">Q×f</span> and <span class="html-italic">ρ<sub>re</sub></span> of BESNPLT HECs; (<b>b</b>) <span class="html-italic">Q×f</span> and packing fraction of BESNPLT HECs.</p>
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<p>(<b>a</b>) TCF and B-site bond valence of BESNPLT HECs; (<b>b</b>) BaLa<sub>2</sub>Ti<sub>4</sub>O<sub>12</sub> (BLT) [<a href="#B36-crystals-14-00754" class="html-bibr">36</a>], BaPr<sub>2</sub>Ti<sub>4</sub>O<sub>12</sub> (BPT) [<a href="#B56-crystals-14-00754" class="html-bibr">56</a>], BaNd<sub>2</sub>Ti<sub>4</sub>O<sub>12</sub> (BNT) [<a href="#B57-crystals-14-00754" class="html-bibr">57</a>], BaSm<sub>2</sub>Ti<sub>4</sub>O<sub>12</sub> (BST) [<a href="#B58-crystals-14-00754" class="html-bibr">58</a>], and BESNPLT ceramics.</p>
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23 pages, 5630 KiB  
Article
MLF-PointNet++: A Multifeature-Assisted and Multilayer Fused Neural Network for LiDAR-UAS Point Cloud Classification in Estuarine Areas
by Yingjie Ren, Wenxue Xu, Yadong Guo, Yanxiong Liu, Ziwen Tian, Jing Lv, Zhen Guo and Kai Guo
Remote Sens. 2024, 16(17), 3131; https://doi.org/10.3390/rs16173131 - 24 Aug 2024
Viewed by 518
Abstract
LiDAR-unmanned aerial system (LiDAR-UAS) technology can accurately and efficiently obtain detailed and accurate three-dimensional spatial information of objects. The classification of objects in estuarine areas is highly important for management, planning, and ecosystem protection. Owing to the presence of slopes in estuarine areas, [...] Read more.
LiDAR-unmanned aerial system (LiDAR-UAS) technology can accurately and efficiently obtain detailed and accurate three-dimensional spatial information of objects. The classification of objects in estuarine areas is highly important for management, planning, and ecosystem protection. Owing to the presence of slopes in estuarine areas, distinguishing between dense vegetation (lawns and trees) on slopes and the ground at the tops of slopes is difficult. In addition, the imbalance in the number of point clouds also poses a challenge for accurate classification directly from point cloud data. A multifeature-assisted and multilayer fused neural network (MLF-PointNet++) is proposed for LiDAR-UAS point cloud classification in estuarine areas. First, the 3D shape features that characterize the geometric characteristics of targets and the visible-band difference vegetation index (VDVI) that can characterize vegetation distribution are used as auxiliary features to enhance the distinguishability of dense vegetation (lawns and trees) on slopes and the ground at the tops of slopes. Second, to enhance the extraction of target spatial information and contextual relationships, the feature vectors output by different layers of set abstraction in the PointNet++ model are fused to form a combined feature vector that integrates low and high-level information. Finally, the focal loss function is adopted as the loss function in the MLF-PointNet++ model to reduce the effect of imbalance in the number of point clouds in each category on the classification accuracy. A classification evaluation was conducted using LiDAR-UAS data from the Moshui River estuarine area in Qingdao, China. The experimental results revealed that MLF-PointNet++ had an overall accuracy (OA), mean intersection over union (mIOU), kappa coefficient, precision, recall, and F1-score of 0.976, 0.913, 0.960, 0.953, 0.953, and 0.953, respectively, for object classification in the three representative areas, which were better than the corresponding values for the classification methods of random forest, BP neural network, Naive Bayes, PointNet, PointNet++, and RandLA-Net. The study results provide effective methodological support for the classification of objects in estuarine areas and offer a scientific basis for the sustainable development of these areas. Full article
(This article belongs to the Special Issue Remote Sensing in Coastal Vegetation Monitoring)
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<p>Study area: (<b>a</b>) location map of the study area; (<b>b</b>) sample area selection range.</p>
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<p>True-color point cloud map of the sample area: (<b>a</b>) Area 1 true-color point cloud; (<b>b</b>) Area 2 true-color point cloud; (<b>c</b>) Area 3 true-color point cloud.</p>
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<p>Sample area category annotation maps: (<b>a</b>) Area 1 category annotation map; (<b>b</b>) Area 2 category annotation map; (<b>c</b>) Area 3 category annotation map.</p>
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<p>Land classification methods for estuarine areas.</p>
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<p>MLF-PointNet++ network architecture.</p>
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<p>Confusion matrix for point cloud classification in estuarine areas via MLF-PointNet++.</p>
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<p>Classification results of the validation area: (<b>a1</b>–<b>c1</b>) show the true labels of the three validation areas; (<b>a2</b>–<b>c2</b>) show the classification results of MLF-PointNet++; and (<b>a3</b>–<b>c3</b>) show the error distributions of the three validation areas. The red boxes represents the misclassified area.</p>
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<p>Comparison of the results of seven classification models in Area 1.</p>
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<p>Comparison of the errors among the seven classification models in Area 1.</p>
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<p>Comparison of the results of seven classification models in Area 2.</p>
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<p>Comparison of the errors among the seven classification models in Area 2.</p>
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<p>Comparison of the results of seven classification models in Area 3.</p>
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<p>Comparison of the errors among the seven classification models in Area 3.</p>
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<p>Classification results for the validation of Area 4.</p>
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<p>Classification results for the validation of Area 5.</p>
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<p>Classification results for the validation of Area 6.</p>
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<p>Error diagram of the classification results for the auxiliary feature ablation experiments: (<b>a1</b>–<b>c1</b>) represent the distributions of the classification errors for M1 in the three validation areas; (<b>a2</b>–<b>c2</b>) are the distributions of the classification errors for M2 in the three validation areas; (<b>a3</b>–<b>c3</b>) represent the distributions of the classification errors for M3 in the three validation areas; and (<b>a4</b>–<b>c4</b>) represent the error distributions of the M4 classification results in the three validation areas.</p>
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<p>Error diagram of the classification results of the loss function ablation experiment: (<b>a1</b>–<b>c1</b>) represent the error distributions of the M5 classification results in the three validation areas; (<b>a2</b>–<b>c2</b>) represent the error distributions of the M6 classification results in the three validation areas; (<b>a3</b>–<b>c3</b>) represent the error distributions of the M7 classification results in the three validation areas; (<b>a4</b>–<b>c4</b>) represent the error distributions of the M8 classification results in the three validation areas; (<b>a5</b>–<b>c5</b>) represent the error distributions of the M9 classification results in the three validation areas.</p>
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11 pages, 1475 KiB  
Article
The Long-Term Monitoring of Atmospheric Polychlorinated Dibenzo-p-Dioxin Dibenzofurans at a Background Station in Taiwan during Biomass Burning Seasons in El Niño and La Niña Events
by Shih Yu Pan, Yen-Shun Hsu, Yuan Cheng Hsu, Tuan Hung Ngo, Charles C.-K. Chou, Neng-Huei Lin and Kai Hsien Chi
Atmosphere 2024, 15(8), 1002; https://doi.org/10.3390/atmos15081002 - 20 Aug 2024
Viewed by 322
Abstract
To measure the long-range transport of PCDD/Fs, a background sampling site at Mt. Lulin station (Taiwan) was selected based on meteorological information and its location relative to burning events in Southeast Asia. During regular sampling periods, a higher concentration of PCDD/Fs was recorded [...] Read more.
To measure the long-range transport of PCDD/Fs, a background sampling site at Mt. Lulin station (Taiwan) was selected based on meteorological information and its location relative to burning events in Southeast Asia. During regular sampling periods, a higher concentration of PCDD/Fs was recorded in 2008 at Mt. Lulin station during La Niña events, with levels reaching 390 fg I-TEQ/m3. In contrast, a higher concentration of 483 fg I-TEQ/m3 was observed in 2013 during biomass burning events. This indicates that La Niña affects the ambient PCDD/F concentrations. The ratio of ΣPCDD/ΣPCDF was 0.59, suggesting significant long-range transport contributions from 2007 to 2023. From 2007 to 2015, the predominant species was 2,3,4,7,8-PCDF, accounting for 25.3 to 39.6% of the total PCDD/Fs. From 2018 onward, 1,2,3,7,8-PCDD became more dominant, accounting for 15.0 to 27.1%. According to the results from the receptor model PMF (n = 150), the sources of PCDD/Fs were identified as dust storms and monsoon events (19.3%), anthropogenic activity (28.5%), and biomass burning events (52.2%). The PSCF values higher than 0.7 highlighted potential PCDD/F emission source regions for Mt. Lulin during biomass burning events, indicating high PSCF values in southern Thailand, Cambodia, and southern Vietnam. Full article
(This article belongs to the Special Issue Toxicity of Persistent Organic Pollutants and Microplastics in Air)
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<p>The locations of the high-altitude sampling site (Mt. Lulin) in Taiwan.</p>
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<p>The distribution of WSIs during El Niño and La Niña.</p>
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<p>Atmospheric PCDD/Fs, PAHs and total suspended particles measured at Mt. Lulin station during 2007–2023.</p>
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<p>The distribution of atmospheric PCDD/Fs in Mt. Lulin during El Niño and La Niña.</p>
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<p>The map of PSCF value in potential emission source region from 2007 to 2023: (<b>a</b>) backward trajectory. (<b>b</b>) the value of PSCF in ambient PCDD/Fs (n = 150).</p>
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18 pages, 4858 KiB  
Article
Sandwich d/f Heterometallic Complexes [(Ln(hfac)3)2M(acac)3] (Ln = La, Pr, Sm, Dy and M = Co; Ln = La and M = Ru)
by Cristian Grechi, Silvia Carlotto, Massimo Guelfi, Simona Samaritani, Lidia Armelao and Luca Labella
Molecules 2024, 29(16), 3927; https://doi.org/10.3390/molecules29163927 - 20 Aug 2024
Cited by 1 | Viewed by 659
Abstract
Sandwich d/f heterometallic complexes [(Ln(hfac)3)2M(acac)3] (Ln = La, Pr, Sm, Dy and M = Co; Ln = La and M = Ru) were prepared in strictly anhydrous conditions reacting the formally unsaturated fragment [Ln(hfac)3] and [...] Read more.
Sandwich d/f heterometallic complexes [(Ln(hfac)3)2M(acac)3] (Ln = La, Pr, Sm, Dy and M = Co; Ln = La and M = Ru) were prepared in strictly anhydrous conditions reacting the formally unsaturated fragment [Ln(hfac)3] and [M(acac)3] in a 2-to-1 molar ratio. These heterometallic complexes are highly sensitive to moisture. Spectroscopic observation revealed that on hydrolysis, these compounds yield dinuclear heterometallic compounds [Ln(hfac)3M(acac)3], prepared here for comparison purposes only. Quantum mechanical calculations supported, on the one hand, the hypothesis on the geometrical arrangement obtained from ATR-IR and NMR spectra and, on the other hand, helped to rationalize the spontaneous hydrolysis reaction. Full article
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<p>ATR-IR spectra of heterodinuclear [Ln(hfac)<sub>3</sub>Co(acac)<sub>3</sub>] (<b>1Ln</b>) complexes: Ln = La, (black); Pr, (blue); Sm, (green) and Dy (orange).</p>
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<p>The <sup>1</sup>H (<b>left</b>) and <sup>19</sup>F (<b>right</b>) NMR of [La(hfac)<sub>3</sub>Co(acac)<sub>3</sub>], <b>1La</b>.</p>
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<p>ATR-IR spectra of [(Ln(hfac)<sub>3</sub>)<sub>2</sub>Co(acac)<sub>3</sub>] (<b>2Ln</b>): La, (black); Pr, (blue); Sm, (green), Dy, (red).</p>
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<p>ATR-IR spectra of [(La(hfac)<sub>3</sub>)<sub>2</sub>Co(acac)<sub>3</sub>] (black) and [La(hfac)<sub>3</sub>Co(acac)<sub>3</sub>] (blue).</p>
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<p>ATR-IR in the range of 1700–1350 cm<sup>−1</sup>: (i) [(La(hfac)<sub>3</sub>)<sub>2</sub>Co(acac)<sub>3</sub>] (black); (ii) [(La(hfac)<sub>3</sub>)<sub>2</sub>Co(acac)<sub>3</sub>] after 1 h air exposure (blue); (iii) [La(hfac)<sub>3</sub>Co(acac)<sub>3</sub>] (green). Bands increasing (green *) or decreasing (black *) with time are marked.</p>
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<p>The <sup>1</sup>H (<b>left</b>) and <sup>19</sup>F (<b>right</b>) NMR spectra of [(La(hfac)<sub>3</sub>)<sub>2</sub>Co(acac)<sub>3</sub>] in CDCl<sub>3</sub>.</p>
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<p>The <sup>1</sup>H and <sup>19</sup>F NMR spectra of [(La(hfac)<sub>3</sub>)<sub>2</sub>Co(acac)<sub>3</sub>] in CDCl<sub>3</sub>: (i) immediately after preparation (black) and (ii) after a few days (brown).</p>
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<p>Optimized structure of [(La(hfac)<sub>3</sub>)<sub>2</sub>Co(acac)<sub>3</sub>] (<b>2La</b>). The green, grey, cyan, red and violet spheres are F, C, La, O and Co atoms, respectively. H atoms are omitted for clarity. Level of theory: PBE def2/JK, DFT-D3.</p>
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<p>The stability trend, as ∆G values, for the dinuclear [Ln(hfac)<sub>3</sub>Co(acac)<sub>3</sub>] and trinuclear [(Ln(hfac)<sub>3</sub>)<sub>2</sub>Co(acac)<sub>3</sub>] complexes (Ln = La, Pr and Sm). All values are in kcal/mol.</p>
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<p>(<b>Left</b>): ATR-IR of [La(hfac)<sub>3</sub>Ru(acac)<sub>3</sub>] (black) and [La(hfac)<sub>3</sub>Co(acac)<sub>3</sub>] (blue). (<b>Right</b>): ATR-IR of [(La(hfac)<sub>3</sub>)<sub>2</sub>Ru(acac)<sub>3</sub>] (black) and [La(hfac)<sub>3</sub>Ru(acac)<sub>3</sub>] (blue).</p>
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<p>ATR-IR of (i) [(La(hfac)<sub>3</sub>)<sub>2</sub>Ru(acac)<sub>3</sub>] (black); (ii) after 4′ of air exposure (blue); (iii) after 30′ (green) and [La(hfac)<sub>3</sub>Ru(acac)<sub>3</sub>] (red). Bands increasing (red *) or decreasing (black *) with time are marked.</p>
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<p>Numbering of <sup>1</sup>H NMR protons in [Ln(hfac)<sub>3</sub>M(acac)<sub>3</sub>].</p>
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<p>Numbering of <sup>1</sup>H NMR protons in [(La(hfac)<sub>3</sub>)<sub>2</sub>Co(acac)<sub>3</sub>].</p>
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<p>Heterodinuclear metal complex formation, exemplified here for a formally unsaturated [Ln(hfac)<sub>3</sub>] and [M(acac)<sub>3</sub>] with the metal M in an octahedral <span class="html-italic">tris</span>-chelate environment.</p>
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<p>Heterotrinuclear f-d-f sandwich metal complex formation, exemplified here for Ln = La and M = Co or Ru in an octahedral <span class="html-italic">tris</span>-chelate environment.</p>
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22 pages, 7839 KiB  
Article
Allele-Specific Hormone Dynamics in Highly Transgressive F2 Biomass Segregants in Sugarcane (Saccharum spp.)
by Noor-ul Ain, Habiba and Ray Ming
Plants 2024, 13(16), 2247; https://doi.org/10.3390/plants13162247 - 13 Aug 2024
Viewed by 469
Abstract
Sugarcane holds global promise as a biofuel feedstock, necessitating a deep understanding of factors that influence biomass yield. This study unravels the intricate dynamics of plant hormones that govern growth and development in sugarcane. Transcriptome analysis of F2 introgression hybrids, derived from the [...] Read more.
Sugarcane holds global promise as a biofuel feedstock, necessitating a deep understanding of factors that influence biomass yield. This study unravels the intricate dynamics of plant hormones that govern growth and development in sugarcane. Transcriptome analysis of F2 introgression hybrids, derived from the cross of Saccharum officinarum “LA Purple” and wild Saccharum robustum “MOL5829”, was conducted, utilizing the recently sequenced allele-specific genome of “LA Purple” as a reference. A total of 8059 differentially expressed genes were categorized into gene models (21.5%), alleles (68%), paralogs (10%), and tandemly duplicated genes (0.14%). KEGG analysis highlighted enrichment in auxin (IAA), jasmonic acid (JA), and abscisic acid (ABA) pathways, revealing regulatory roles of hormone repressor gene families (Aux/IAA, PP2C, and JAZ). Signaling pathways indicated that downregulation of AUX/IAA and PP2C and upregulation of JAZ repressor genes in high biomass segregants act as key players in influencing downstream growth regulatory genes. Endogenous hormone levels revealed higher concentrations of IAA and ABA in high biomass, which contrasted with lower levels of JA. Weighted co-expression network analysis demonstrated strong connectivity between hormone-related key genes and cell wall structural genes in high biomass genotypes. Expression analysis confirmed the upregulation of genes involved in the synthesis of structural carbohydrates and the downregulation of inflorescence and senescence-related genes in high biomass, which suggested an extended vegetative growth phase. The study underscores the importance of cumulative gene expression, including gene models, dominant alleles, paralogs, and tandemly duplicated genes and activators and repressors of disparate hormone (IAA, JA, and ABA) signaling pathways are the points of hormone crosstalk in contrasting biomass F2 segregants and could be applied for engineering high biomass acquiring varieties. Full article
(This article belongs to the Section Plant Genetics, Genomics and Biotechnology)
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<p>Partial least squares-discriminant analysis (PLS-DA), Hierarchical clustering, and volcano plot of DEGs: (<b>A</b>) PLS-DA score plot of FPKM data of DEGs generated by using preprocessed original data shows the clustering of extreme biomass segregants. Component 1 (26%) clearly distinguishes the two biomass groups. (<b>B</b>) (<b>a</b>) Hierarchical clustering heatmap of the DEGs based on FPKM expression values, (<b>b</b>) Six clusters indicating the up and down regulated genes identified in heatmap. (<b>C</b>) Volcano plot overall shows the range of log2FC of DEGs, i.e., 24 to −26 from right to left.</p>
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<p>KEGG and GO analysis of DEGs: (<b>A</b>) Bar plot for the KEGG categories and enriched DEGs in KEGG analysis. The abscissa depicts enriched pathways while gene number is plotted on an ordinate axis. (<b>B</b>) GO shows the enrichment of potential DEGs in different categories, i.e., biological processes, cellular components, and molecular function.</p>
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<p>Heatmap of log2FC values in HB segregants. Red and black values in legend show down and upregulated genes. (<b>A</b>) shows ubiquitin-mediated signaling of auxin and jasmonic acid signaling pathways. (<b>B</b>) depicts the expression patterns of hormone-responsive growth-related genes in cell wall and terminal developmental phases, i.e., inflorescence and senescence. <span class="html-italic">S-40</span>: Senescence regulator, <span class="html-italic">ATHB</span>: ARABIDOPSIS THALIANA HOMEOBOX 7, <span class="html-italic">KNAT1</span>: BREVIPEDICELLUS, <span class="html-italic">PHOX2/ARIX</span>: Transcription factor PHOX2/ARIX, <span class="html-italic">ELF3</span>: EARLY FLOWERING 3 (<span class="html-italic">ELF3</span>), <span class="html-italic">FPF1</span>: FLOWERING PROMOTING FACTOR 1, <span class="html-italic">AGAMOUS-LIKE 12</span>: AGAMOUS-LIKE 12, <span class="html-italic">XXT</span>: galactosyl transferase GMA12/MNN10 family, <span class="html-italic">EXPANSIN</span>: EXPANSIN, <span class="html-italic">CESA</span>: cellulose synthase subfamily, <span class="html-italic">GAE</span>: GDP-mannose 4,6 dehydratase, <span class="html-italic">CSLA02</span>: belongs to the glycosyltransferase 2 family, <span class="html-italic">XTH</span>: Xyloglucan endohydrolysis (<span class="html-italic">XEH</span>) and or endotransglycosylation (<span class="html-italic">XET</span>), and <span class="html-italic">UGT</span>: UDP-glycosyltransferase family.</p>
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<p>Weighted gene co-expression network analysis (WGCNA) of DEGs between HB and LB segregants. (<b>A</b>) eigengene modules, module–trait relationship, and module–module relationship. Module–trait heatmap shows Pearson’s correlation of all the modules with samples, whereas in the module–module relationship, progressive saturation in blue and red color points to high co-expression interconnectedness. Additionally, it shows dendrogram of module clustering, with green and red horizontal lines representing threshold (0.25, 0.3). (<b>B</b>) Cytoscape network shows co-expression network of blue module, which depicts highly upregulated genes in HB segregants (<b>C</b>) Cystoscape network of overrepresented DEGs in module “greenyellow”. Size and color of nodes are proportional to weights, whereas edge colors correspond to module names.</p>
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<p>Heatmap generated using log2FC. Expression dynamics of TFs and PKs involved in high biomass samples. Green and blue scales represent up and downregulated TFs in HB samples, whereas grey color indicates blanks.</p>
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<p>(<b>A</b>) Concentrations of the endogenous hormones, i.e., ABA, JA, and IAA, in the leaves of high and low biomass F2 segregants. Bar charts present the means with error bars showing standard errors, different letters are based on one-way ANOVA and LSD tests at α = 0.05. (<b>B</b>) Linear regression model of hormone content and qPCR values of the genes identified in RNA-Seq. analysis in the respective signaling pathway.</p>
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<p>Confirmation of FPKM by qPCR expression. Green bars represent the relative expression in qPCR, and orange lines represent FPKM values in transcriptome for corresponding genes. Values on the <span class="html-italic">y</span>-axis indicate relative expression levels of qPCR (<b>left</b>) and RNA-Seq (<b>right</b>). Error bars show standard error of the means at (<span class="html-italic">p</span> &lt; 0.05), and “r” is indicative of correlation between qPCR and FPKM expression values.</p>
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26 pages, 5132 KiB  
Article
Microbial Diversity of Soil in a Mediterranean Biodiversity Hotspot: Parque Nacional La Campana, Chile
by Carolina Quinteros-Urquieta, Jean-Pierre Francois, Polette Aguilar-Muñoz, Roberto Orellana, Rodrigo Villaseñor, Andres Moreira-Muñoz and Verónica Molina
Microorganisms 2024, 12(8), 1569; https://doi.org/10.3390/microorganisms12081569 - 31 Jul 2024
Viewed by 605
Abstract
Parque Nacional La Campana (PNLC) is recognized worldwide for its flora and fauna, rather than for its microbial richness. Our goal was to characterize the structure and composition of microbial communities (bacteria, archaea and fungi) and their relationship with the plant communities typical [...] Read more.
Parque Nacional La Campana (PNLC) is recognized worldwide for its flora and fauna, rather than for its microbial richness. Our goal was to characterize the structure and composition of microbial communities (bacteria, archaea and fungi) and their relationship with the plant communities typical of PNLC, such as sclerophyllous forest, xerophytic shrubland, hygrophilous forest and dry sclerophyllous forest, distributed along topoclimatic variables, namely, exposure, elevation and slope. The plant ecosystems, the physical and chemical properties of organic matter and the soil microbial composition were characterized by massive sequencing (iTag-16S rRNA, V4 and ITS1-5F) from the DNA extracted from the soil surface (5 cm, n = 16). A contribution of environmental variables, particularly related to each location, is observed. Proteobacteria (35.43%), Actinobacteria (32.86%), Acidobacteria (10.07%), Ascomycota (76.11%) and Basidiomycota (15.62%) were the dominant phyla. The beta diversity (~80% in its axes) indicates that bacteria and archaea are linked to their plant categories, where the xerophytic shrub stands out with the most particular microbial community. More specifically, Crenarchaeote, Humicola and Mortierella were dominant in the sclerophyllous forest; Chloroflexi, Cyanobacteria and Alternaria in the xerophytic shrubland; Solicoccozyma in the dry sclerophyllous forest; and Cladophialophora in the hygrophilous forest. In conclusion, the structure and composition of the microbial consortia is characteristic of PNLC’s vegetation, related to its topoclimatic variables, which suggests a strong association within the soil microbiome. Full article
(This article belongs to the Special Issue Advances in Soil Microbial Ecology)
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<p>Map that shows PNLC sampling locations (1, 2, 3, 4). Vegetation map, adapted from Hauck et al. [<a href="#B38-microorganisms-12-01569" class="html-bibr">38</a>].</p>
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<p>Graphical representation of the four sample sites characterized by their exposure and elevation.</p>
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<p>Physicochemical variables of the soil samples: (<b>a</b>) C/N. Significant C/N differences were observed between the hygrophilous forest and the xerophytic shrubland, and between the dry sclerophyllous forest and the xerophytic shrubland. (<b>b</b>) pH. No significant differences between locations were observed. (<b>c</b>) Dry density (g/cm<sup>3</sup>). Significant differences were observed between the hygrophilous forest and the xerophytic shrubland (<b>d</b>) d15N and d13C isotope content.</p>
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<p>Redundancy analysis (RDA) that shows the variability of the environmental conditions associated with the plant communities. (For further information, see <a href="#app1-microorganisms-12-01569" class="html-app">Supplementary Table S3</a>.)</p>
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<p>(<b>a</b>) Prokaryote and (<b>b</b>) fungi soil microorganism’s alpha diversity in PNLC plant communities. Grey dots correspond to replicate outliers.</p>
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<p>PCoA (PERMANOVA <span class="html-italic">p</span>-value &lt; 0.05) that illustrates the microbial variability of the soil of the plant communities at the phylum level of (<b>a</b>) prokaryotes and (<b>b</b>) fungi. The blue arrows represent the environmental parameters that significantly correlate with the microbial communities (envfit, <span class="html-italic">p</span>-value &lt; 0.05).</p>
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<p>PCoA (PERMANOVA <span class="html-italic">p</span>-value &lt; 0.05) that illustrates the microbial variability of the soil of the plant communities at the phylum level of (<b>a</b>) prokaryotes and (<b>b</b>) fungi. The blue arrows represent the environmental parameters that significantly correlate with the microbial communities (envfit, <span class="html-italic">p</span>-value &lt; 0.05).</p>
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<p>Venn diagram that shows the comparison of ASVs richness associated with (<b>a</b>) prokaryotes and (<b>b</b>) fungi, grouped on the basis of the soil of the plant communities. Sclerophilous forest—purple area, Xerophytic shrubland—yellow area, Hygrophilous forest—green area, Dry sclerophilous forest—orange area.</p>
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<p>The box plot shows the relative abundance of bacterial and archaeal phyla in the PNLC soil categorized by the different plant communities.</p>
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<p>The box plots show the relative abundance of the fungal phyla in the PNLC soil categorized by different plant communities.</p>
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<p>Heatmap of the 18 most abundant genera of prokaryotes in the soil (<span class="html-italic">n</span> = 16) analyzed in PNLC, categorized by plant communities.</p>
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<p>Heatmap for the 20 genera of fungi in the soil (<span class="html-italic">n</span> = 16) analyzed in PNLC categorized by plant communities.</p>
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<p>Volcano plot that shows the ASVs of microorganisms (bacteria, archaea and fungi) when comparing the xerophytic shrubland vs. sclerophyllous forest plant communities in PNLC. The red line indicates the significantly different ASVs (<span class="html-italic">p</span>-adj). Complete data in <a href="#app1-microorganisms-12-01569" class="html-app">Supplementary Table S4</a>.</p>
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<p>Volcano plot that shows the ASVs of microorganism when comparing the hygrophilous forest and dry sclerophyllous forest plant communities in PNLC. The red line indicates the significantly different ASVs (<span class="html-italic">p</span>-adj). Complete data in <a href="#app1-microorganisms-12-01569" class="html-app">Supplementary Table S5</a>.</p>
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<p>Predicted potential microbial function profile by comparison of the 4 study sites. (<b>a</b>) Prokaryotes, (<b>b</b>) fungi.</p>
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30 pages, 3349 KiB  
Article
Predictive and Prognostic 18F-Fluorocholine PET/CT Radiomics Nomogram in Patients with Castration-Resistant Prostate Cancer with Bone Metastases Treated with 223Ra
by Marcos Cruz-Montijano, Mariano Amo-Salas, Javier Cassinello-Espinosa, Iciar García-Carbonero, Jose Carlos Villa-Guzman and Ana Maria Garcia-Vicente
Cancers 2024, 16(15), 2695; https://doi.org/10.3390/cancers16152695 - 29 Jul 2024
Viewed by 697
Abstract
Purpose: We aimed to develop a nomogram able to predict treatment failure, skeletal events, and overall survival (OS) in patients with castration-resistant prostate cancer with bone metastases (CRPC-BM) treated with Radium-223 dichloride (223Ra). Patients and Methods: Patients from the Castilla-La Mancha [...] Read more.
Purpose: We aimed to develop a nomogram able to predict treatment failure, skeletal events, and overall survival (OS) in patients with castration-resistant prostate cancer with bone metastases (CRPC-BM) treated with Radium-223 dichloride (223Ra). Patients and Methods: Patients from the Castilla-La Mancha Spanish region were prospectively included in the ChoPET-Rad multicenter study from January 2015 to December 2022. Patients underwent baseline, interim, and end-of-treatment bone scintigraphy (BS) and 18F-Fluorocholine PET/CT (FCH PET/CT) scans, obtaining multiple imaging radiomics as well as clinical and biochemical variables during follow-up and studying their association with the previously defined end-points. Survival analysis was performed using the Kaplan–Meier method and Cox regression. Multivariate logistic and Cox regression models were calculated, and these models were depicted by means of nomograms. Results: Median progression-free survival (PFS) and OS were 4 and 14 months (mo), respectively. The variables that showed independent and significant association with therapeutic failure were baseline alkaline phosphatase (AP) levels (p = 0.022) and the characteristics of BM on the CT portion of PET/CT (p = 0.017). In the case of OS, the significant variables were therapeutic failure (p = 0.038), the number of lines received after 223Ra (p < 0.001), average SUVmax (p = 0.002), bone marrow infiltration in FCH PET/CT (p = 0.006), and interim FCH PET/CT response (p = 0.048). Final nomograms included these variables, showing good discrimination among the 100 patients included in our study. In the study of skeletal events, only OS showed a significant association in the multivariate analysis, resulting in an inconsistent nomogram design. Conclusions: FCH PET/CT appears to be a good tool for evaluating patients eligible for treatment with 223Ra, as well as for their follow-up. Thus, findings derived from it, such as the morphological characteristics of BM in the CT, bone marrow infiltration, or the response to 223Ra in the interim study, have proven to be solid and useful variables in the creation of nomograms for predicting therapeutic failure and OS. Full article
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<p>Kaplan–Meier OS curves of baseline AP levels (<b>upper left panel</b>), LDH levels (<b>upper right panel</b>), and ECOG performance status (<b>lower panel</b>).</p>
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<p>Kaplan–Meier OS curves of baseline FCH PET/CT radiomics: high tumor burden (<b>upper left panel</b>), uptake higher than liver for the most hypermetabolic bone metastases (<b>upper right panel</b>), bone marrow involvement (<b>lower left panel</b>), and soft tissue involvement (<b>lower right panel</b>).</p>
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<p>Kaplan–Meier OS curves of binary response in interim FCH PET/CT scans (<b>left panel</b>) and BS (<b>right panel</b>).</p>
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<p>Designed nomograms to predict therapeutic failure (<b>left panel</b>) and the probability of survival at 12 and 24 months (<b>right panel</b>). The nomograms were developed based on the 100 patients in our population with CRPC-BM who received <sup>223</sup>Ra therapy. To obtain the probability of therapeutic failure and survival, the values for each variable of the patients included in each nomogram are marked. Then, a straight vertical line is drawn up to the “Points” line at the top of the nomogram. This determines how many points are attributed to each variable. Once this is performed for each variable, the sum of all the points obtained is calculated and added to the “Total Points” line at the bottom of the nomogram. This value is then used to assess the individual probability of predicting the risk of therapeutic failure (<b>left panel</b>) and survival at 12 and 24 months (<b>right panel</b>).</p>
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<p>Patient ≠ 1. A 71-year-old man, diagnosed with prostate adenocarcinoma Gleason Score 9 (4 + 5), underwent prostatectomy followed by adjuvant radiotherapy to the surgical bed due to early PSA progression. Three years after diagnosis, bone metastases were detected, prompting the start of systemic treatment lines (Enzalutamide and Docetaxel). <sup>223</sup>Ra was administered as the third line. At the start of treatment, the patient was in good general condition (ECOG 0), experienced pain, and had baseline PSA levels of 50.7 ng/dL, AP of 377 IU/L, and LDH of 377 IU/L. Baseline BS (<b>A</b>) shows polymetastatic disease (&gt;20 lesions) affecting both the axial and extra-axial skeleton. Baseline FCH PET/CT (<b>B</b>) shows the presence of mixed characteristic BM, bone marrow infiltration, and uptake of the most hypermetabolic BM higher than liver and soft tissue involvement at the pelvic lymph nodes. The concordance between both studies was moderate, defining FCH PET/CT more BM with respect to BS.</p>
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<p>Patient ≠ 1. PSA and AP showed a steady increase after <sup>223</sup>Ra initiation. Clinical deterioration was observed after the third <sup>223</sup>Ra administration. Interim BS (<b>A</b>) and FCH PET/CT (<b>B</b>) show disease progression. The patient died 6 months after starting treatment with <sup>223</sup>Ra.</p>
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<p>Patient ≠ 2. An 88-year-old man diagnosed with metastatic prostate cancer, Gleason Score 7 (4 + 3), from the onset. He received first-line treatment with Enzalutamide until biochemical and bone progression after 5 years. Treatment with <sup>223</sup>Ra was proposed as second-line. Baseline BS and FCH PET/CT (<b>A</b>) show oligometastatic disease with only axial involvement and predominantly osteoblastic. At the start of <sup>223</sup>Ra treatment, the patient was in very good general condition (ECOG 0) with baseline PSA levels of 14.9 ng/mL, AP of 73 IU/L, and LDH of 450 IU/L. The patient completed 6 doses of <sup>223</sup>Ra, remaining stable in the interim BS and FCH PET/CT studies (<b>B</b>) but showed bone progression at the end of treatment in both the BS and FCH PET/CT (<b>C</b>). His PSA levels increased during the treatment, while AP levels remained stable. After <sup>223</sup>Ra treatment, the patient received Docetaxel and experienced a bone event that consisted of a painful metastatic bone at 17 months, treated with palliative vertebral radiotherapy. He ultimately died with an OS of 30 months.</p>
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<p>Patient ≠ 3. A 69-year-old man diagnosed with prostate cancer, Gleason score 8 (4 + 4) stage IV T3N1M1 (hepatic, pulmonary, nodal, and bone infiltration). The patient achieved a complete radiologic response after receiving first-line Docetaxel with an anti-androgen blockade. After progression, Abiraterone–Prednisone was administered with a partial biochemical response. <sup>223</sup>Ra was administered as the third line. Baseline BS and FCH PET/CT (<b>A</b>) show oligometastatic axial and extra-axial BM on the BS and polymetastatic (6–20 lesions) axial and extra-axial involvement on the FCH PET/CT without soft tissue involvement, with some BM showing lytic characteristics and higher uptake than the liver. Despite this, the concordance between both techniques was good, with metabolic activity predominating over osteogenic. At the start of treatment, the patient was in very good general condition (ECOG 0) with pain and baseline PSA values of 5 ng/dL, AP of 87 IU/L, and LDH of 363 IU/L. He received 6 doses of <sup>223</sup>Ra, with PSA values progressing from the start of treatment, although AP and LDH values did not. The interim BS study shows rib deposits interpreted as bone progression while the FCH PET/CT showed stability (<b>B</b>)<b>,</b> explained by the interpretation of rib deposits as probable fractures. However, the end-treatment scans (<b>C</b>) show bone and nodal progression of FCH PET/CT with stability in the BS. Subsequently, treatment with Abiraterone–Prednisone was resumed, followed shortly after by Cabazitaxel. The patient died 10 months after <sup>223</sup>Ra initiation.</p>
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