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17 pages, 9855 KiB  
Article
A Rod-like Bi2O3 Photocatalyst Derived from Bi-Based MOFs for the Efficient Adsorption and Catalytic Reduction of Cr(VI)
by Qin Fang, Luying Chen, Qiucheng Fu, Yongjuan Chen, Jiao He, Liang Jiang, Zhiying Yan and Jiaqiang Wang
Int. J. Mol. Sci. 2024, 25(23), 13052; https://doi.org/10.3390/ijms252313052 - 4 Dec 2024
Abstract
Heavy metal ion pollution poses a serious threat to the natural environment and human health. Photoreduction through Bi-based photocatalysts is regarded as an advanced green technology for solving environmental problems. However, their photocatalytic activity is limited by the rapid recombination of photogenerated e [...] Read more.
Heavy metal ion pollution poses a serious threat to the natural environment and human health. Photoreduction through Bi-based photocatalysts is regarded as an advanced green technology for solving environmental problems. However, their photocatalytic activity is limited by the rapid recombination of photogenerated e and h+ pairs and a low photo-quantum efficiency. In this work, an optimal precursor of Bi-based MOFs was identified by using different solvents, and rod-like Bi2O3 materials were derived by in situ oxidation of Bi atoms in the precursor. The adsorption and photocatalytic reduction efficiency of the prepared Bi2O3 materials for Cr(VI) were evaluated under visible light irradiation. The results showed that the prepared materials had a large specific surface area and enhanced visible light absorption. Bi2O3(DMF/MeOH-3)-400 had a large specific surface area and many active adsorption sites, and it had the highest adsorption of Cr(VI) (49.13%) among the materials. Bi2O3(DMF/MeOH-3)-400 also had the highest photocatalytic reduction efficiency, and it achieved 100% removal of 10 mg·L−1 Cr(VI) within 90 min under light. In addition, the material showed remarkable stability after three consecutive photocatalytic cycles. The enhanced photocatalytic performance was mainly attributed to the fast separation of electron–hole pairs and efficient electron transfer in the MOF-derived materials, which was confirmed by electrochemical tests and PL spectroscopy. Reactive species trapping experiments confirmed that electrons were the main active substances; accordingly, a possible photocatalytic mechanism was proposed. In conclusion, this work provides a new perspective for designing novel photocatalysts that can facilitate the removal of Cr(VI) from water. Full article
(This article belongs to the Special Issue Properties and Applications of Nanoparticles and Nanomaterials)
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Figure 1

Figure 1
<p>XRD patterns of Bi-BTC precursors and the as-prepared samples at 300 °C, 400 °C, and 500 °C (<b>a</b>), and using different solvents (<b>b</b>).</p>
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<p>SEM images: (<b>a</b>) Bi-BTC(DMF/MeOH-3), (<b>b</b>) Bi<sub>2</sub>O<sub>3</sub>(DMF/MeOH-3)-400, (<b>c</b>) Bi<sub>2</sub>O<sub>3</sub>(DMF)-400, and (<b>d</b>) Bi<sub>2</sub>O<sub>3</sub>(MeOH)-400.</p>
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<p>(<b>a</b>–<b>c</b>) TEM images of Bi<sub>2</sub>O<sub>3</sub>(DMF/MeOH-3)-400, and (<b>d</b>) HRTEM image of Bi<sub>2</sub>O<sub>3</sub> (DMF/MeOH-3)-400.</p>
Full article ">Figure 4
<p>N<sub>2</sub> adsorption–desorption isotherms were used to obtain the specific surface area and pore structure of Bi-BTC (<b>a</b>) and the as-prepared samples (<b>b</b>).</p>
Full article ">Figure 5
<p>XPS spectra of Bi<sub>2</sub>O<sub>3</sub>(DMF/MeOH-3)-400: (<b>a</b>) survey scan, (<b>b</b>) Bi 4f, (<b>c</b>) O 1s, and (<b>d</b>) C 1s.</p>
Full article ">Figure 6
<p>FT−IR spectrum of the Bi<sub>2</sub>O<sub>3</sub>(DMF/MeOH-3)-400 samples.</p>
Full article ">Figure 7
<p>(<b>a</b>) UV−Vis DRS spectra and (<b>b</b>) Tauc plots of as-prepared samples.</p>
Full article ">Figure 8
<p>Photocatalytic reduction performance for Cr(VI) by the as-prepared samples and commercial Bi<sub>2</sub>O<sub>3</sub> (<b>a</b>). The corresponding apparent first−order reaction kinetics curve (<b>b</b>).</p>
Full article ">Figure 9
<p>Performance of the photocatalyst in the reduction of Cr(VI) at different initial pH values (<b>a</b>). The corresponding apparent first−order reaction kinetics curve (<b>b</b>).</p>
Full article ">Figure 10
<p>(<b>a</b>) Reusability of Bi<sub>2</sub>O<sub>3</sub>(DMF/MeOH-3)-400 in the photocatalytic reduction of Cr(VI). Comparison of (<b>b</b>) XRD patterns, (<b>c</b>) XPS survey before and after Bi<sub>2</sub>O<sub>3</sub>(DMF/MeOH-3)-400 was used. (<b>d</b>) Cr 2p spectra after Bi<sub>2</sub>O<sub>3</sub>(DMF/MeOH-3)-400 was used.</p>
Full article ">Figure 11
<p>(<b>a</b>) Steady−state PL spectra, (<b>b</b>) EIS Nyquist plots, and (<b>c</b>) TPR plots of prepared Bi<sub>2</sub>O<sub>3</sub> and commercial Bi<sub>2</sub>O<sub>3</sub>. (<b>d</b>) Mott−Schottky plots of prepared Bi<sub>2</sub>O<sub>3</sub> at 500, 1000, 1500 Hz.</p>
Full article ">Figure 12
<p>Reactive species trapping experiments of Bi<sub>2</sub>O<sub>3</sub>(DMF/MeOH-3)-400.</p>
Full article ">Scheme 1
<p>Synthetic route of the rod−like Bi<sub>2</sub>O<sub>3</sub> samples.</p>
Full article ">Scheme 2
<p>Proposed photocatalytic mechanism of Bi<sub>2</sub>O<sub>3</sub>(DMF/MeOH−3)-400.</p>
Full article ">
18 pages, 6357 KiB  
Article
Liposomal and Nanostructured Lipid Nanoformulations of a Pentacyclic Triterpenoid Birch Bark Extract: Structural Characterization and In Vitro Effects on Melanoma B16-F10 and Walker 256 Tumor Cells Apoptosis
by Dumitriţa Rugină, Mihai Adrian Socaciu, Madalina Nistor, Zorita Diaconeasa, Mihai Cenariu, Flaviu Alexandru Tabaran and Carmen Socaciu
Pharmaceuticals 2024, 17(12), 1630; https://doi.org/10.3390/ph17121630 - 4 Dec 2024
Abstract
Background/Objectives: Pentacyclic triterpenoids are increasingly studied as anticancer agents with many advantages compared to synthetic chemotherapeutics. The aim of this study was to prepare liposomal and nanostructured lipid formulations including a standardized extract of silver birch (Betula pendula) outer bark [...] Read more.
Background/Objectives: Pentacyclic triterpenoids are increasingly studied as anticancer agents with many advantages compared to synthetic chemotherapeutics. The aim of this study was to prepare liposomal and nanostructured lipid formulations including a standardized extract of silver birch (Betula pendula) outer bark (TTs) and to evaluate their potential as anticancer agents in vitro, using Melanoma B16-F10 and Walker carcinoma cells. Methods: Appropriate solvents were selected for efficient TTs extraction, and original recipes were used to obtain Pegylated liposomes and nanolipid complexes with entrapped TTs, comparative to pure standards (betulinic acid and doxorubicin) in similar conditions. The composition, morphology, and sizes of all nanoformulations were checked by high-performance liquid chromatography/mass spectrometry, Transmission Electronic Microscopy, and Diffraction Light Scattering. The entrapment efficiency and its impact on cell viability, cell cycle arrest, and apoptosis by flow cytometry was also measured on both cancer cell lines. Conclusions: The standardized TTs, including betulin, lupeol, and betulinic acid, showed good stability and superior activity compared to pure betulinic acid. According to experimental data, TTs showed good entrapment in liposomal and NLC nanoformulations, both delivery systems including natural, biodegradable ingredients and enhanced bioavailability. The apoptosis and necrosis effects were more pronounced for TTs liposomal formulations in both types of cancer cells, with lower cytotoxicity compared to Doxorubicin, and can be correlated with their increased bioavailability. Full article
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Figure 1

Figure 1
<p>Most representative pentacyclic lupane-type triterpenoids in birch bark extracts: structure, molecular weight [MW], and maximal UV absorption.</p>
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<p>Morphology of Lipo and NLC nanoformulations containing AB, TTs, and Doxo, comparative to controls, as determined by TEM. Size distribution diameters (mean ± SD) (nm) of the formulations, the encapsulation efficiency (EE% ± SD), and the PDI values are included. The final concentrations (C<sub>f</sub>) for each nanoformulation including AB, TT, and Doxo were mentioned.</p>
Full article ">Figure 3
<p>(<b>a</b>–<b>l</b>) The flow cytometry distribution of cell cycle affected in Melanoma B16-F10 cell population. (<b>a</b>) Non-treated cells. (<b>b</b>,<b>c</b>) After incubation with Lipo and NLC controls. (<b>d</b>–<b>f</b>) After incubation with AB, TTs, and Doxo. (<b>g</b>–<b>i</b>) After incubation with Lipo-AB, Lipo-TTs and Lipo-Doxo. (<b>j</b>–<b>l</b>) After incubation with NLC-AB, NLC-TTs, NLC-Doxo.</p>
Full article ">Figure 3 Cont.
<p>(<b>a</b>–<b>l</b>) The flow cytometry distribution of cell cycle affected in Melanoma B16-F10 cell population. (<b>a</b>) Non-treated cells. (<b>b</b>,<b>c</b>) After incubation with Lipo and NLC controls. (<b>d</b>–<b>f</b>) After incubation with AB, TTs, and Doxo. (<b>g</b>–<b>i</b>) After incubation with Lipo-AB, Lipo-TTs and Lipo-Doxo. (<b>j</b>–<b>l</b>) After incubation with NLC-AB, NLC-TTs, NLC-Doxo.</p>
Full article ">Figure 4
<p>(<b>a</b>–<b>l</b>) The flow cytometry distribution of cell cycle affected Walker 256 cacinoma cell population. (<b>a</b>) Non-treated cells. (<b>b</b>,<b>c</b>) After incubation with Lipo and NLC controls. (<b>d</b>–<b>f</b>) After incubation with AB, TTs, and Doxo. (<b>g</b>–<b>i</b>) After incubation with Lipo-AB, Lipo-TTs and Lipo-Doxo. (<b>j</b>–<b>l</b>) After incubation with NLC-AB, NLC-TTs, and NLC-Doxo.</p>
Full article ">Figure 4 Cont.
<p>(<b>a</b>–<b>l</b>) The flow cytometry distribution of cell cycle affected Walker 256 cacinoma cell population. (<b>a</b>) Non-treated cells. (<b>b</b>,<b>c</b>) After incubation with Lipo and NLC controls. (<b>d</b>–<b>f</b>) After incubation with AB, TTs, and Doxo. (<b>g</b>–<b>i</b>) After incubation with Lipo-AB, Lipo-TTs and Lipo-Doxo. (<b>j</b>–<b>l</b>) After incubation with NLC-AB, NLC-TTs, and NLC-Doxo.</p>
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<p>Comparative effects of Lipo- and NLC-nanoformulations and free extracts (TTs, AB and Doxo) on Melanoma B16-F10 (<b>A</b>) and Walker 256 (<b>B</b>) cell cycle.</p>
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<p>Comparative results of Annexin V/propidium iodide-stained flow cytometry for Melanoma B16F10 (<b>a</b>,<b>c</b>,<b>e</b>,<b>g</b>) and Walker 256 cells (<b>b</b>,<b>d</b>,<b>f</b>,<b>h</b>) after 24 h of incubation. (<b>a</b>) Melanoma B16F10 untreated cells; (<b>b</b>) Walker 256 untreated cells; (<b>c</b>,<b>e</b>,<b>g</b>) Melanoma B16F10 incubated with AB, TTs and Doxo; (<b>d</b>,<b>f</b>,<b>h</b>) Walker 256 cells incubated with AB, TTs, and Doxo. Q1 + Q2 = A (early and late apoptosis); Q3—viable cells; Q4—necrosis.</p>
Full article ">Figure 6 Cont.
<p>Comparative results of Annexin V/propidium iodide-stained flow cytometry for Melanoma B16F10 (<b>a</b>,<b>c</b>,<b>e</b>,<b>g</b>) and Walker 256 cells (<b>b</b>,<b>d</b>,<b>f</b>,<b>h</b>) after 24 h of incubation. (<b>a</b>) Melanoma B16F10 untreated cells; (<b>b</b>) Walker 256 untreated cells; (<b>c</b>,<b>e</b>,<b>g</b>) Melanoma B16F10 incubated with AB, TTs and Doxo; (<b>d</b>,<b>f</b>,<b>h</b>) Walker 256 cells incubated with AB, TTs, and Doxo. Q1 + Q2 = A (early and late apoptosis); Q3—viable cells; Q4—necrosis.</p>
Full article ">Figure 7
<p>Histograms showing the comparative percentages of apoptotic cells ((<b>A</b>) melanoma B16-F10; (<b>B</b>) Walker 256) and necrotic cells ((<b>C</b>) melanoma B16-F10; (<b>D</b>) Walker 256) after the treatment with different Lipo- and NLC-formulations comparative to free molecules Doxo, AB, and TTs controls. A = Q1 + Q2 (early and late apoptosis); Q4—necrosis.</p>
Full article ">
15 pages, 14323 KiB  
Article
Synergistic Effect of Sono-Photocatalysis on the Degradation of Rhodamine B Dye Using BiFeO3 Nanoparticles Synthesized via a Modified Combustion Method
by Anel I. Robles-Cortes, Daniel Flores-Ramírez, Lissette Medina-de la Rosa, Denisse F. González-Ramírez, José Ortiz-Landeros and Issis C. Romero-Ibarra
Ceramics 2024, 7(4), 1880-1894; https://doi.org/10.3390/ceramics7040118 (registering DOI) - 4 Dec 2024
Abstract
Water contamination has become a global concern, and the prevalence of complex substances known as emerging contaminants constitute a risk to human health and the environment. This work focused on an innovative approach of integrating sonolysis and photocatalysis to remove a standard textile [...] Read more.
Water contamination has become a global concern, and the prevalence of complex substances known as emerging contaminants constitute a risk to human health and the environment. This work focused on an innovative approach of integrating sonolysis and photocatalysis to remove a standard textile dye efficiently. A highly photo-active, bismuth ferrite (BiFeO3) nanocatalyst with single particle sizes between 86 and 265 nm was obtained by a novel one-pot combustion method using a deep eutectic solvent as a precursor. The said catalyst was thoroughly characterized and evaluated for photocatalytic and sono-photocatalytic degradation of rhodamine B (RhB). Photocatalytic experiments were conducted under visible light irradiation (450–600 nm). Sono-photocatalytic (SPC) experiments were conducted, focusing on the influence of operational parameters (frequency, power, and pH) on the degradation performance. High-frequency values of 578, 866, and 1138 kHz were explored to promote cavitation dynamics and reactive species generation, improving removal efficiency. Results demonstrated that when sonolysis and photocatalysis were performed separately, the degradation efficiency ranged between 85 and 87%. Remarkably, when the combined SPC degradation was carried out, the RhB removal reached about 99.9% after 70 min. It is discussed that this behavior is due to the increased generation of OH radicals as a product of the cavitation phenomena related to the ultrasound-assisted process. Moreover, it is argued that SPC significantly improves reaction kinetics and mass transfer rates, facilitating catalyst dispersion and contact with the RhB molecules. Finally, the stability of the catalyst was evaluated in five repeated RhB removal cycles, where the activity remained consistently strong. Full article
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Graphical abstract

Graphical abstract
Full article ">Figure 1
<p>(<b>a</b>) TGA-DTA curve and (<b>b</b>) X-ray diffraction pattern of the as-synthesized sponge-like BiFeO<sub>3</sub> sample at 550 °C for 2.5 h.</p>
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<p>(<b>a</b>) SEM image of the as-synthesized BiFeO<sub>3</sub>, (<b>b</b>) energy dispersive spectroscopy (EDS), (<b>c</b>) TEM image of the BiFeO<sub>3,</sub> wherein circle illustrates lattice fringes, and (<b>d</b>) particle size distribution.</p>
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<p>(<b>a</b>) Adsorption–desorption isotherm; inset shows the pore size distribution. (<b>b</b>) Typical determination of the bandgap (<span class="html-italic">E<sub>g</sub></span>); the calculated <span class="html-italic">E<sub>g</sub></span> values are within the visible light absorption range of 590 nm. (Insert shows UV-Vis NIR spectrum).</p>
Full article ">Figure 4
<p>(<b>a</b>) XPS spectrum of the BiFeO<sub>3</sub> catalyst. Core levels of (<b>b</b>) Bi 4f, (<b>c</b>) Fe 2p, and (<b>d</b>) O 1s.</p>
Full article ">Figure 5
<p>Effect of the operating parameters on the sono-photocatalytic degradation of RhB. (<b>a</b>) Effect of frequency, (<b>b</b>) effect of pH, (<b>c</b>) zeta potential values as a function of pH, and (<b>d</b>) effect of ultrasonic power. All experiments were conducted at 25 °C, but the parameters varied.</p>
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<p>UV-vis spectra of RhB collected during the sono-photocatalytic degradation of Rhodamine B at 866 kHz, pH = 3, 80% power (insert shows dye discoloration over time).</p>
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<p>(<b>a</b>) RhB degradation rates under photolysis, sonolysis, photocatalysis, and sono-photocatalysis and (<b>b</b>) BiFeO<sub>3</sub> reusability experiments over five consecutive cycles.</p>
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<p>Characterization of the catalyst used for five consecutive cycles: (<b>a</b>) XRD pattern and inset (<b>b</b>) SEM image.</p>
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<p>Schematic illustration of the possible sono-photocatalytic mechanism occurring in the BiFeO<sub>3</sub> catalyst under light irradiation.</p>
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13 pages, 13135 KiB  
Article
PER1 Oscillation in Rat Parathyroid Hormone and Calcitonin Producing Cells
by Birgitte Georg, Henrik L. Jørgensen and Jens Hannibal
Int. J. Mol. Sci. 2024, 25(23), 13006; https://doi.org/10.3390/ijms252313006 - 3 Dec 2024
Viewed by 258
Abstract
Many endocrine glands exhibit circadian rhythmicity, but the interplay between the central circadian clock in the suprachiasmatic nucleus (SCN), the peripheral endocrine clock, and hormones is sparsely understood. We therefore studied the cellular localizations of the clock protein PER1, parathyroid hormone (PTH) and [...] Read more.
Many endocrine glands exhibit circadian rhythmicity, but the interplay between the central circadian clock in the suprachiasmatic nucleus (SCN), the peripheral endocrine clock, and hormones is sparsely understood. We therefore studied the cellular localizations of the clock protein PER1, parathyroid hormone (PTH) and calcitonin (CT) in the parathyroid and thyroid glands, respectively. Thyroid glands, including the parathyroids, were dissected at different time-points from rats housed in 12 h:12 h light–darkness cycles, and were double-immunostained for PER1 and PTH or CT. Sera were analyzed for PTH, CT, phosphate, and calcium. In both glands, PER1 expression peaked late at night, while limited staining was seen during the daytime. High-resolution microscopy revealed cytosolic PER1 at zeitgeber time (ZT)12, and nucleic staining at ZT24 in both PTH and CT cells. PTH peaked at Z12–ZT16, while neither CT staining nor serum CT oscillated during the daily cycle. Serum PTH was significantly higher at ZT12 than ZT24, but only phosphate was found to exhibit significant diurnal oscillation. The staining of the calcium-sensitive receptor (CSR) did not demonstrate circadian oscillation. In conclusion, PER1 expression peaked late at night/early in the morning in hormone-producing cells of both the thyroid and parathyroid glands. In the parathyroids, this was preceded by a PTH peak, while neither CT nor CRS were found to oscillate. Full article
(This article belongs to the Special Issue Molecular Advances in Circadian Rhythm and Metabolism)
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Figure 1

Figure 1
<p>Daily oscillation of PER1 immunostaining in rat parathyroid glands. Confocal images of PER1 staining of glands taken at different time points during a 12 h:12 h light–dark cycle. Glands of four animals at each timepoint were stained, and representative pictures are shown. (<b>a</b>) The presence of PER1 in a parathyroid gland taken at ZT2; (<b>b</b>) at midday (ZT6), very little PER1 staining is seen; (<b>c</b>,<b>d</b>) there is almost an absence of staining in glands from ZT10 and ZT12; (<b>e</b>,<b>f</b>) at ZT14 and ZT16, more evident staining is appearing; (<b>g</b>,<b>h</b>) intense PER1 staining in parathyroids from ZT20 and ZT24. Scale bars: 25 µm.</p>
Full article ">Figure 2
<p>Intracellular distribution of PER1 rat parathyroid glands during the daily cycle. High-power confocal images of PER1 immunostaining (green) and DAPI nuclear staining (red) at selected time points during a 12 h:12 h light–dark cycle. (<b>a</b>) At ZT2, PER1 is primarily localized in the cell nuclei; (<b>b</b>) the primarily cytoplasmic PER1 localization at ZT12; (<b>c</b>) at ZT14, PER1 is seen to appear in the nuclei; (<b>d</b>) at ZT20, PER1 is almost entirely present in the cell nuclei. Scale bars: 10 µm.</p>
Full article ">Figure 3
<p>Co-localization of PER1 and PTH in the same cells. (<b>a</b>) Immunostaining of PER1 in green and PTH in red in the same cells; (<b>b</b>) DAPI staining in blue, confirming the nucleic localization of PER1, now appearing in turquoise. Scale bars: 10 µm.</p>
Full article ">Figure 4
<p>Results of in situ hybridization (ISH) detecting PTH mRNA during the 12 h:12 h light–dark cycle. (<b>a</b>) Graphical illustration of the semi-quantification of ISH results showing higher PTH mRNA expression at the beginning and early in the night (ZT12-ZT14) than late at night (n = 2–4); the difference was, however, not statistically significant; (<b>b</b>) representative photomicrograph of PTH ISH at ZT10; (<b>c</b>) ISH of PTH mRNA at ZT14. Scale bars: 25 µm.</p>
Full article ">Figure 5
<p>Confocal images of immunofluorescent PTH staining of rat parathyroid glands taken during a 12 h:12 h light–dark cycle. (<b>a</b>,<b>b</b>): No or very low staining in early morning and midday; (<b>c</b>,<b>d</b>): Intense staining in early night; (<b>e</b>,<b>f</b>): Decreasing staining in late night. Scale bares: 25 µm.</p>
Full article ">Figure 6
<p>Confocal images of PER1 (green) and calcitonin (red) double immunofluorescence staining of the thyroid gland taken during a 12 h:12 h light–dark cycle. PER1 is seen to vary in both intensity and localization during the daily cycle. (<b>b</b>) shows absence in the day; (<b>d</b>–<b>f</b>) increasing PER1 staining in the night; (<b>f</b>) Shows nucleic localization at dawn. Scale bars: 25 µm.</p>
Full article ">Figure 7
<p>Confocal images of calcitonin immunostaining of thyroid glands taken during a 12 h:12 h light–dark cycle. Although differing, diurnal oscillation in calcitonin staining intensity was not found. Scale bares: 25 µm.</p>
Full article ">Figure 8
<p>Confocal microphotographs of CSR immunostaining of the parathyroid and thyroid glands. (<b>a</b>) Picture of both glands taken at ZT10; (<b>b</b>) picture of both glands taken ZT20; (<b>c</b>) higher magnification of the parathyroid gland showing CSR in green and DAPI in red; (<b>d</b>) higher magnification of CSR (green) and DAPI (red); (<b>e</b>) detailed high-resolution analysis in and 3D reconstruction (x, Y, Z) showing the cell membrane localization of CSR. Scale bars: (<b>a</b>,<b>b</b>): 50 µm; (<b>c</b>): 20 µm; (<b>d</b>): 25 µm; (<b>e</b>): 8 µm.</p>
Full article ">Figure 9
<p>Graphical illustration of serum concentrations of PTH (<b>a</b>), calcitonin (<b>b</b>), phosphate (<b>c</b>), and calcium (<b>d</b>) during a 12 h:12 h LD cycle. PTH and calcitonin were measured by commercial ELISA kits, and phosphate and calcium concentrations were measured on Cobas 8000. Only the concentration of phosphate displayed significant 24 h sinusoidal oscillation. The concentration of PTH at ZT12 was, however, significantly higher (<span class="html-italic">p</span> &lt; 0.05) than at ZT24. NS: Not Significant.</p>
Full article ">
15 pages, 5941 KiB  
Article
Lighting Patterns Regulate Flowering and Improve the Energy Use Efficiency of Calendula Cultivated in Plant Factories with Artificial Lighting
by Maitree Munyanont, Na Lu, Dannisa Fathiya Rachma, Thanit Ruangsangaram and Michiko Takagaki
Agriculture 2024, 14(12), 2208; https://doi.org/10.3390/agriculture14122208 - 3 Dec 2024
Viewed by 302
Abstract
Calendula is an edible flower with highly beneficial bioactive compounds for human health. Environmental factors such as light influence flower yield and quality. Calendula is cultivated under controlled environments in plant factories with artificial lighting (PFALs), which enhance its productivity. However, electricity is [...] Read more.
Calendula is an edible flower with highly beneficial bioactive compounds for human health. Environmental factors such as light influence flower yield and quality. Calendula is cultivated under controlled environments in plant factories with artificial lighting (PFALs), which enhance its productivity. However, electricity is the main operating cost, with fees based on the time of use in some countries. This study aimed to investigate the effects of lighting patterns on calendula growth and yield. Two varieties of calendula seedlings were cultivated in a PFAL and subjected to six different lighting patterns, i.e., 6 h/6 h, 12 h/12 h, 6 h/2 h, and 18 h/6 h (light/dark) and two continuous lighting patterns with varying light intensities (24 h-200 and 24 h-400). The results indicated that plants cultivated under the 6 h/2 h, 18 h/6 h, 24 h-200, and 24 h-400 conditions showed a more rapid appearance of the first flower bud than those cultivated under the 6 h/6 h and 12 h/12 h conditions. The number of flowers and the fresh and dried weights tended to increase with a longer photoperiod. Interestingly, the total carotenoid content (TCC) of “Citrus Orange” increased under 6 h/6 h and 12 h/12 h conditions compared with the others. For “Orange Gem”, continuous lighting (24 h) increased the total phenolic content (TPC) and 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity in flowers compared with the 6 h/6 h 12 h/12 h, and 6 h/2 h treatments. The energy use efficiency (EUE) under the 24 h-200 condition was the highest in terms of flower yield and secondary metabolite production. These results suggest that lighting patterns can be used to modulate the growth and flowering of calendula and to maximize EUE. Full article
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Figure 1
<p>Morphology of calendula plants grown in a PFAL under different lighting patterns at 55 DAS.</p>
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<p>Plant dry weight (<b>A</b>) and flower-to-shoot ratio (<b>B</b>) of calendula grown in a PFAL under different lighting patterns. Data are shown as the mean ± SD; <span class="html-italic">n</span> = 6. The different letters show significant differences among treatments applied to the same variety based on Tukey’s test (<span class="html-italic">p</span> &lt; 0.05).</p>
Full article ">Figure 2 Cont.
<p>Plant dry weight (<b>A</b>) and flower-to-shoot ratio (<b>B</b>) of calendula grown in a PFAL under different lighting patterns. Data are shown as the mean ± SD; <span class="html-italic">n</span> = 6. The different letters show significant differences among treatments applied to the same variety based on Tukey’s test (<span class="html-italic">p</span> &lt; 0.05).</p>
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<p>Number of days from sowing to the appearance of the first flower bud (<b>A</b>), and the fresh and dry weights of calendula flowers (<b>B</b>,<b>C</b>) obtained from plants grown in a PFAL under different lighting patterns. Data are shown as the mean ± SD; <span class="html-italic">n</span> = 6. The different letters show significant differences among treatments applied to the same variety based on Tukey’s test (<span class="html-italic">p</span> &lt; 0.05).</p>
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<p>Number of days from sowing to the appearance of the first flower bud (<b>A</b>), and the fresh and dry weights of calendula flowers (<b>B</b>,<b>C</b>) obtained from plants grown in a PFAL under different lighting patterns. Data are shown as the mean ± SD; <span class="html-italic">n</span> = 6. The different letters show significant differences among treatments applied to the same variety based on Tukey’s test (<span class="html-italic">p</span> &lt; 0.05).</p>
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<p>Flower diameter (<b>A</b>) and flower redness (<b>B</b>) of calendula plants grown in a PFAL under different lighting patterns. Data are shown as the mean ± SD; <span class="html-italic">n</span> = 6. The different letters show significant differences among treatments applied to the same variety based on Tukey’s test (<span class="html-italic">p</span> &lt; 0.05).</p>
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<p>Yield of calendula (total number of flowers (<b>A</b>), total fresh and dry weights of flowers (<b>B</b>,<b>C</b>)) grown in a PFAL under different lighting patterns. Data are shown as the mean ± SD; <span class="html-italic">n</span> = 6. The different letters show significant differences among treatments applied to the same variety based on Tukey’s test (<span class="html-italic">p</span> &lt; 0.05).</p>
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<p>Yield of calendula (total number of flowers (<b>A</b>), total fresh and dry weights of flowers (<b>B</b>,<b>C</b>)) grown in a PFAL under different lighting patterns. Data are shown as the mean ± SD; <span class="html-italic">n</span> = 6. The different letters show significant differences among treatments applied to the same variety based on Tukey’s test (<span class="html-italic">p</span> &lt; 0.05).</p>
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<p>Total carotenoid content (<b>A</b>), total phenolic content (<b>B</b>), DPPH scavenging activity (<b>C</b>), and the correlation between secondary metabolites and antioxidative activity (<b>D</b>) of calendula grown in a PFAL under different lighting patterns. Data are shown as the mean ± SD; <span class="html-italic">n</span> = 6. The different letters show significant differences among treatments applied to the same variety based on Tukey’s test (<span class="html-italic">p</span> &lt; 0.05).</p>
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<p>Total carotenoid content (<b>A</b>), total phenolic content (<b>B</b>), DPPH scavenging activity (<b>C</b>), and the correlation between secondary metabolites and antioxidative activity (<b>D</b>) of calendula grown in a PFAL under different lighting patterns. Data are shown as the mean ± SD; <span class="html-italic">n</span> = 6. The different letters show significant differences among treatments applied to the same variety based on Tukey’s test (<span class="html-italic">p</span> &lt; 0.05).</p>
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<p>EUE of yield (total number of flowers; (<b>A</b>) total flower fresh weight; (<b>B</b>) and total flower dry weight; (<b>C</b>)), secondary metabolites (total carotenoid content; (<b>D</b>) and total phenolic compound content; (<b>E</b>)) and DPPH scavenging activity (<b>F</b>) of calendula grown in a PFAL under different lighting patterns. Data are shown as the mean ± SD; <span class="html-italic">n</span> = 6. The different letters show significant differences among treatments applied to the same variety based on Tukey’s test (<span class="html-italic">p</span> &lt; 0.05).</p>
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15 pages, 2380 KiB  
Article
Effect of a Broiler-Specific Light Spectrum on Growth Performance and Adrenocortical Activity in Chickens: A Pilot Study on a Commercial Farm
by Livio Galosi, Luca Todini, Laura Menchetti, Annaïs Carbajal, Rupert Palme, Nicola Ruggiero, Roberto Falconi and Alessandra Roncarati
Vet. Sci. 2024, 11(12), 618; https://doi.org/10.3390/vetsci11120618 - 2 Dec 2024
Viewed by 256
Abstract
This study evaluated the effect of a broiler-specific light spectrum on productive performance corticosterone (fCC) and androgen dehydroepiandrosterone (fDHEA) concentrations in feathers, and glucocorticoid (GCMs) and androgen (AMs) metabolites in droppings of broilers. Two groups of female Ross 308 broilers were reared under [...] Read more.
This study evaluated the effect of a broiler-specific light spectrum on productive performance corticosterone (fCC) and androgen dehydroepiandrosterone (fDHEA) concentrations in feathers, and glucocorticoid (GCMs) and androgen (AMs) metabolites in droppings of broilers. Two groups of female Ross 308 broilers were reared under white LED (WL, n = 9000) and broiler-specific LED (BSL, n = 9000) lights. The body weight (BW) of 150 randomly selected animals/groups was measured weekly. Droppings and feathers were collected at the end of the cycle (29 days) from 20 animals/group. The BSL group showed higher final BW than WL (1407 ± 11 vs. 1341 ± 15 g, respectively; p < 0.001) and higher indices of uniformity (76.8% vs. 61.2% animals in the 10% around the mean, respectively; p < 0.001). No difference between groups was found in fCC and fDHEA concentrations or in the fCC–fDHEA, indicating similar long-term HPA axis activity during the cycle. A higher concentration of GCMs was found in the BSL group, indicating higher glucocorticoid secretion before sampling, with neither a difference in AMs nor in GCMs–AMs. Finally, there was a positive correlation between fCC and fDHEA and between GCMs and AMs (p < 0.01). Our findings suggest that the use of broiler-specific light improved the productivity performances of chickens without long-term consequences on HPA activation. However, the results of this pilot study in a commercial farm setting must be interpreted with caution and need confirmation. Full article
(This article belongs to the Section Veterinary Physiology, Pharmacology, and Toxicology)
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<p>Poultry houses used in this trial. (<b>A</b>). Poultry house WL, equipped with compact fluorescent white light. (<b>B</b>). Poultry house BSL equipped with broiler-specific light (NatureDynamics System, ONCE by Signify, Philips).</p>
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<p>Body weight (BW) changes in the white LED (WL) and broiler-specific LED (BSL) groups. Values are means ± standard errors. The asterisks indicate significant differences between the two groups for each time point (*** <span class="html-italic">p</span> &lt; 0.001; n.s. not significant). T1 = 8 days; T2 = 15 days; T3 = 22 days; T4 = 29 days.</p>
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<p>Corticosterone (<b>A</b>) and DHEA (<b>B</b>) in feathers in the white LED (WL) and broiler-specific LED (BSL) groups. Values are means ± standard errors (n.s. = not significant). DHEA = dehydroepiandrosterone.</p>
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<p>Pyramid chart and test statistics for the Mann–Whitney test comparing the fCC/fDHEA (expressed as pg/mm) distribution of white LED (WL) and broiler-specific LED (BSL) groups. Numbers on the x-axis indicate the frequency of the number of animals, while numbers on the Y-axis indicate the value of the fCC–fDHEA ratio. fCC = corticosterone concentrations in feathers; fDHEA = dehydroepiandrosterone concentrations in feathers.</p>
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<p>Glucocorticoid (<b>A</b>) and androgen (<b>B</b>) metabolites in droppings in the white LED (WL) and broiler-specific LED (BSL) groups. Values are means ± standard errors. The asterisk indicates significant differences between the two groups (* <span class="html-italic">p</span> &lt; 0.05; n.s. = not significant).</p>
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<p>Pyramid chart and test statistics for the Mann–Whitney test comparing the distribution of glucocorticoid/androgen metabolites ratio (GCMs/AMs) in the droppings of white LED (WL) and broiler-specific LED (BSL) groups. Numbers on the x-axis indicate the frequency of the number of animals, while numbers on the Y-axis indicate the value of the ratio.</p>
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14 pages, 8507 KiB  
Article
Magnetic Ternary Hybrid Composites as an Efficient Photocatalyst for Degradation of Acid Orange 7 Dye
by Yaohui Xu, Qin Wang, Yuting Li and Zhao Ding
Catalysts 2024, 14(12), 880; https://doi.org/10.3390/catal14120880 - 2 Dec 2024
Viewed by 249
Abstract
Based on the photocatalytic activity and magnetic nature of magnetite and goethite, as well as the oxygen storage characteristic of cerianite, a magnetic ternary hybrid composite including cubic CeO2, cubic Fe3O4 and orthorhombic FeOOH, designated as Fe3 [...] Read more.
Based on the photocatalytic activity and magnetic nature of magnetite and goethite, as well as the oxygen storage characteristic of cerianite, a magnetic ternary hybrid composite including cubic CeO2, cubic Fe3O4 and orthorhombic FeOOH, designated as Fe3O4/FeOOH/CeO2, was successfully synthesized with different Ce:Fe molar ratios using a simple hydrothermal route without subsequent calcination process, and employed as photocatalysts for the degradation of Acid Orange 7 (AO7) dye. The absorption range of light by the Fe3O4/FeOOH/CeO2 composites was broadened, and the intensity was enhanced. Furthermore, there existed a possibility of hybridization and doping among the three crystalline structures, with the elements Ce, Fe and O exhibiting a uniform distribution, significantly enhancing the photocatalytic efficiency of the Fe3O4/FeOOH/CeO2 composites in promoting the photodegradation of AO7. The magnetic response behaviors of hybrid composites synthesized with different Ce:Fe molar ratios were investigated. The adsorptive degradation of AO7 in darkness and the photocatalytic degradation of AO7 under UV light illumination were evaluated. Moreover, ten cycling runs of the photocatalytic degradation of AO7 under simulated UV illumination of Fe3O4/FeOOH/CeO2 synthesized with a Ce:Fe molar ratio of 1:15 were performed. The hybrid ternary composites were proved to have excellent magnetic sensitivity, exhibited outstanding photocatalytic activities and demonstrated remarkable stability. It is anticipated that magnetic Fe3O4/FeOOH/CeO2 ternary hybrid composites may have potential applications in the treatment of organic dye sewage. Full article
(This article belongs to the Special Issue Novel Catalysts for Environmental Catalysis)
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<p>XRD patterns of samples synthesized with different molar ratios of Ce:Fe (1:1, 1:2, 1:5, 1:10, 1:15 and 1:20), and two reference samples synthesized without Fe and without Ce.</p>
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<p>SEM images of reference samples (<b>a</b>) CeO<sub>2</sub> and (<b>b</b>) Fe<sub>3</sub>O<sub>4</sub>; composites synthesized with Ce:Fe molar ratios of (<b>c</b>) 1:1, (<b>d</b>) 1:2, (<b>e</b>) 1:5, (<b>f</b>) 1:10, (<b>g</b>) 1:15 and (<b>h</b>) 1:20.</p>
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<p>(<b>a</b>) Selected area in the SEM micrograph and elemental mappings of (<b>b</b>) Fe (rad), (<b>c</b>) Ce (green) and (<b>d</b>) O (blue) for the Fe<sub>3</sub>O<sub>4</sub>/FeOOH/CeO<sub>2</sub> composite synthesized with Ce:Fe molar ratio of 1:15.</p>
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<p>Magnetic response behaviors of CeO<sub>2</sub>, Fe<sub>3</sub>O<sub>4</sub> and the composites synthesized with Ce:Fe molar ratios of 1:1, 1:2, 1:5, 1:10, 1:15 and 1:20. [samples] = 0.1 g; <span class="html-italic">V</span> = 30 mL. In each picture, the vial on the left, standing upright, shows the suspension after 30 min of ultrasonic dispersion followed by 24 h of settling. The vial on the right, with a magnet, shows the result of 2 min magnetic separation.</p>
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<p>Magnetic response behaviors of CeO<sub>2</sub>, Fe<sub>3</sub>O<sub>4</sub> and the composites synthesized with Ce:Fe molar ratios of 1:1, 1:2, 1:5, 1:10, 1:15 and 1:20. [samples] = 0.1 g; <span class="html-italic">V</span> = 30 mL. In each picture, the top vial shows the suspension after 30 min of ultrasonic dispersion, while the bottom vial with a magnet displays 2 min of magnetic separation.</p>
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<p>UV–VIS absorption spectra of samples synthesized with different molar ratios of Ce:Fe (1:1, 1:2, 1:5, 1:10, 1:15 and 1:20), and two reference samples synthesized without Fe and without Ce.</p>
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<p>Photo-degradation rates of AO7 in the presence of CeO<sub>2</sub>, Fe<sub>3</sub>O<sub>4</sub> and the Fe<sub>3</sub>O<sub>4</sub>/FeOOH/CeO<sub>2</sub> composites synthesized with Ce:Fe molar ratios of 1:1, 1:2, 1:5, 1:10, 1:15 and 1:20. [catalyst] = 0.1 g; [AO7] = 5 mg/L, <span class="html-italic">V</span> = 100 mL; ambient temperature = 25 °C; without pH preadjustment.</p>
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<p>Cycling runs in the photocatalytic degradation of AO7 under simulated UV illumination of the Fe<sub>3</sub>O<sub>4</sub>/FeOOH/CeO<sub>2</sub> composite synthesized with a Ce:Fe molar ratio of 1:15.</p>
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<p>The synthesis of Fe<sub>3</sub>O<sub>4</sub>/FeOOH/CeO<sub>2</sub> magnetic complex catalysts, along with two reference samples: CeO<sub>2</sub> and Fe<sub>3</sub>O<sub>4</sub>.</p>
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18 pages, 2165 KiB  
Article
Towards Bio-Crude Refinery Integration: Hydrodeoxygenation and Co-Hydroprocessing with Light Cycle Oil
by Athanasios Dimitriadis and Stella Bezergianni
Energies 2024, 17(23), 6032; https://doi.org/10.3390/en17236032 - 30 Nov 2024
Viewed by 336
Abstract
Hydrothermal liquefaction of solid waste has been gaining more and more attention over the last few years. However, the properties of the HTL product, i.e., biocrude, are limiting its direct utilization. As a result, HTL biocrude upgrading is essential to improve its quality. [...] Read more.
Hydrothermal liquefaction of solid waste has been gaining more and more attention over the last few years. However, the properties of the HTL product, i.e., biocrude, are limiting its direct utilization. As a result, HTL biocrude upgrading is essential to improve its quality. The main objective of the current research is to study the hydrotreatment stabilization of HTL biocrude, produced from spent coffee grounds, utilizing commercial hydrotreated catalysts, and also to investigate the integration of the stabilized biocrude into a light cycle oil (LCO) hydrotreatment plant for coprocessing to target hybrid fuel production. The results have shown that hydrotreatment is a very promising technology that can successfully remove the oxygen content from raw biocrude by hydrodeoxygenation, decarbonylation and decarboxylation reactions, leading to a stabilized product. The stabilized product can be easily blended with the LCO stream of a typical refinery, leading to the production of jet and diesel boiling range hydrocarbons, favoring at the same time the hydrogen consumption of the process. The findings of this manuscript set the basis for future research targeting the production of renewable advanced biofuels from HTL biocrude from municipal waste. Full article
(This article belongs to the Special Issue New Trends in Biofuels and Bioenergy for Sustainable Development II)
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<p>GC-MS analysis of HTL biocrude oil (<b>A</b>) shows the composition of the feed and (<b>B</b>) shows the carboxylic acid concentration.</p>
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<p>Distillation curves from raw HTL biocrude and from the stabilized products from conditions 1, 2 and 3.</p>
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<p>GC-MS analysis from raw HTL biocrude (feed) and from the stabilized products from conditions 1, 2 and 3.</p>
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<p>Aliphatic hydrocarbon analysis from the products of the examined conditions.</p>
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<p>Process mass yields.</p>
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<p>Coprocessing testing overview of product sulfur content and the corresponding reactor temperature variation during coprocessing.</p>
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<p>Product distillation curve from neat LCO, 10% and 20% <span class="html-italic">v</span>/<span class="html-italic">v</span> blend with stabilized biocrude.</p>
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<p>ΔP plot during coprocessing of LCO and stabilized biocrude blends.</p>
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16 pages, 4601 KiB  
Article
Optimization of the Mechanical Recycling of Phenolic Resins for Household Appliances
by Francesco Valentini, Daniele Rigotti, Matteo Saletti, Alberto Beccaro, Laura Pasquardini, Alessandro Pegoretti and Andrea Dorigato
Polymers 2024, 16(23), 3378; https://doi.org/10.3390/polym16233378 - 30 Nov 2024
Viewed by 362
Abstract
In light of the significant impact of climate change, it is imperative to identify effective solutions to reduce the environmental burdens of industrial production and to promote recycling strategies also for thermosetting polymers. In this work, the mechanical recycling of phenolic resins, obtained [...] Read more.
In light of the significant impact of climate change, it is imperative to identify effective solutions to reduce the environmental burdens of industrial production and to promote recycling strategies also for thermosetting polymers. In this work, the mechanical recycling of phenolic resins, obtained from industrial production scrap of plastic knobs for household appliances, was optimized. The feasibility of a partial substitution of virgin materials with recycled ones was investigated both at a laboratory and industrial scale. Finally, the environmental benefits arising from the use of recycled material were quantified through a life cycle assessment (LCA). The results of laboratory characterization demonstrated that the thermal properties of the phenolic resins were not influenced by the presence of recycled material, and the mechanical performances were not significantly impaired up to a recycled content of 30 wt%. The industrial production trials demonstrated the feasibility of replacing up to 15 wt% of virgin material without any influence on the aesthetical features of the produced components. Finally, LCA of industrially produced knobs highlighted a limited benefit of virgin material substitution in the case of novolac chromium-plated samples, while an overall environmental impact reduction of around 7–10% was detected in the case of resol-based materials. Full article
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<p>Example of injection moulding sprue used as recyclate in the mechanical recycling process.</p>
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<p>Production scheme for the samples tested at lab scale.</p>
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<p>Pictures of the prepared samples (NOV30) using recycled material: (<b>a</b>) after compression moulding, and (<b>b</b>) after CNC milling.</p>
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<p>Production scheme for the industrial production trials.</p>
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<p>SEM micrographs of the cryofractured surface of (<b>a</b>) NOV, (<b>b</b>) NOV30CR, (<b>c</b>) RES, and (<b>d</b>) RES30 samples. Recycled particles are highlighted by arrows.</p>
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<p>TGA curves of (<b>a</b>) novolac and (<b>b</b>) resol samples containing different amounts of recycled material.</p>
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<p>DSC curves (first heating scan) of novolac (<b>a</b>) and resol (<b>b</b>) samples.</p>
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<p>Representative flexural stress–strain curves of (<b>a</b>) novolac and (<b>b</b>) resol-based samples containing different amounts of recycled material.</p>
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<p>Flexural stress–strain curves of novolac samples produced using recycled material subjected to multiple reprocessing cycles.</p>
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<p>Representative images of knobs produced upon industrial production trials based on (<b>a</b>) NOV and NOV15 samples, and (<b>b</b>) RES and RES15 samples.</p>
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<p>Group analysis on the life cycle impact assessment results of the industrial production of a knob based on (<b>a</b>) NOV and (<b>b</b>) RES samples.</p>
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32 pages, 10673 KiB  
Article
Light-Modulated Circadian Synaptic Plasticity in the Somatosensory Cortex: Link to Locomotor Activity
by Małgorzata Jasińska, Ewa Jasek-Gajda, Marek Ziaja, Jan A. Litwin, Grzegorz J. Lis and Elżbieta Pyza
Int. J. Mol. Sci. 2024, 25(23), 12870; https://doi.org/10.3390/ijms252312870 - 29 Nov 2024
Viewed by 230
Abstract
The circadian clock controls various physiological processes, including synaptic function and neuronal activity, affecting the functioning of the entire organism. Light is an important external factor regulating the day–night cycle. This study examined the effects of the circadian clock and light on synaptic [...] Read more.
The circadian clock controls various physiological processes, including synaptic function and neuronal activity, affecting the functioning of the entire organism. Light is an important external factor regulating the day–night cycle. This study examined the effects of the circadian clock and light on synaptic plasticity, and explored how locomotor activity contributes to these processes. We analyzed synaptic protein expression and excitatory synapse density in the somatosensory cortex of mice from four groups exposed to different lighting conditions (LD 12:12, DD, LD 16:8, and LL). Locomotor activity was assessed through individual wheel-running monitoring. To explore daily and circadian changes in synaptic proteins, we performed double-immunofluorescence labeling and laser scanning confocal microscopy imaging, targeting three pairs of presynaptic and postsynaptic proteins (Synaptophysin 1/PSD95, Piccolo/Homer 1, Neurexins/PICK1). Excitatory synapse density was evaluated by co-labeling presynaptic and postsynaptic markers. Our results demonstrated that all the analyzed synaptic proteins exhibited circadian regulation modulated by light. Under constant light conditions, only Piccolo and Homer 1 showed rhythmicity. Locomotor activity was also associated with the circadian clock’s effects on synaptic proteins, showing a stronger connection to changes in postsynaptic protein levels. Excitatory synapse density peaked during the day/subjective day and exhibited an inverse relationship with locomotor activity. Continued light exposure disrupted cyclic changes in synapse density but kept it consistently elevated. These findings underscore the crucial roles of light and locomotor activity in regulating synaptic plasticity. Full article
(This article belongs to the Special Issue Synapse Dynamics: From Molecular Mechanisms to Functional Plasticity)
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<p>Analysis of daily and circadian rhythmicity. (<b>A</b>) <span class="html-italic">Tau</span>—period of daily/circadian rhythm [h]; (<b>B</b>) <span class="html-italic">Delta</span>—shift (phase advance or phase delay) of the activity onset [h]; (<b>C</b>) <span class="html-italic">Alpha</span>—duration of activity phase [h]; (<b>D</b>) <span class="html-italic">Rho</span>—duration of rest phase; (<b>E</b>) overall activity [wheel revolutions/day]; (<b>F</b>) Qp—robustness of circadian rhythm [%]; (<b>G</b>) night/subjective night activity [wheel revolutions/day]; (<b>H</b>) day/subjective day activity [wheel revolutions/day]; and (<b>I</b>) percentage of activity during the day/subjective day to total activity [%]. The experimental light conditions: light–dark 12 h:12 h (LD 12:12), constant darkness (DD), long photoperiod LD 16 h:8 h (LD 16:8), and constant light (LL). The graphs show means ± SEM (one-way ANOVA; *** <span class="html-italic">p</span> &lt; 0.001 and ** <span class="html-italic">p</span> &lt; 0.01). The asterisks located directly above the bars signify that the difference applies to all groups.</p>
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<p>Daily and circadian locomotor activity. The left panel (<b>A</b>–<b>D</b>) shows representative double-plotted actograms of mice running-wheel activity under different conditions. The right panel (<b>E</b>–<b>H</b>) displays representative periodograms (average waveform of activity) showing mouse locomotor activity as measured over 10 days under different conditions. The time points at which the mice were sacrificed are indicated by the arrows—yellow arrow if in light, black if in darkness. The experimental light conditions: light–dark 12 h:12 h (LD 12:12), constant darkness (DD), long photoperiod LD 16 h:8 h (LD 16:8), and constant light (LL).</p>
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<p>Daily and circadian changes in the expression of presynaptic proteins. The left panel shows the number of presynaptic protein immunopuncta and the area covered by protein (<b>A</b>,<b>B</b>) Synaptophysin 1, (<b>C</b>,<b>D</b>) Piccolo, (<b>E</b>,<b>F</b>) Neurexin 1/2/3, throughout the day–night or the subjective day–night cycle under different conditions. The graphs show means ± SEM (one-way ANOVA; *** <span class="html-italic">p</span> &lt; 0.001, ** <span class="html-italic">p</span> &lt; 0.01, * <span class="html-italic">p</span> &lt; 0.05). The asterisks located directly above the bars signify that the difference applies to all time points within the group. The right panel displays images illustrating the immunopositive reaction for respective presynaptic proteins, highlighting the minimum and maximum values observed in different conditions where statistically significant differences were observed between individual time points: (<b>G</b>) Synaptophysin 1, (<b>H</b>) Piccolo, (<b>I</b>) Neurexin 1/2/3. Scale bar, 10 µm. The experimental light conditions are as follows: light–dark 12 h:12 h (LD), constant darkness (DD), prolonged light LD 16 h:8 h (LD16), and constant light (LL). Time points are defined as follows: 0—ZT0/CT0: the beginning of the day/subjective day; 6—ZT6/CT6: the middle of the day/subjective day; 12—ZT12/CT12: the beginning of the night/subjective night; 18—ZT18/CT18: the middle of the night/subjective night.</p>
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<p>Daily and circadian changes in the expression of postsynaptic proteins. The left panel shows the number of postsynaptic protein immunopuncta and the area covered by protein (<b>A</b>,<b>B</b>) PSD95, (<b>C</b>,<b>D</b>) Homer 1, and (<b>E</b>,<b>F</b>) PICK1 throughout the day–night or the subjective day–night cycle under different conditions. The graphs show means ± SEM (one-way ANOVA; *** <span class="html-italic">p</span> &lt; 0.001, ** <span class="html-italic">p</span> &lt; 0.01, * <span class="html-italic">p</span> &lt; 0.05). The asterisks located directly above the bars signify that the difference applies to all time points within the group. The right panel displays images illustrating the immunopositive reaction for respective presynaptic proteins, highlighting the minimum and maximum values observed in different conditions where statistically significant differences were observed between individual time points: (<b>G</b>) PSD95, (<b>H</b>) Homer 1, (<b>I</b>) PICK1. Scale bar, 10 µm. The experimental light conditions: light–dark 12 h:12 h (LD), constant darkness (DD), prolonged light LD 16 h:8 h (LD16), and constant light (LL). Time points are defined as follows: 0—ZT0/CT0: the beginning of the day/subjective day; 6—ZT6/CT6: the middle of the day/subjective day; 12—ZT12/CT12: the beginning of the night/subjective night; 18—ZT18/CT18: the middle of the night/subjective night.</p>
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<p>Daily and circadian changes in excitatory synapses. The left panel shows the density of excitatory synapses throughout the day–night or the subjective day–night cycle under different conditions. The synaptic protein pairs are (<b>A</b>) Synaptophysin 1—PSD95 (Syp1+/PSD95+), (<b>B</b>) Piccolo—Homer 1 (Pic+/Hom1+), and (<b>C</b>) Neurexin 1/2/3—PICK1 (NRXN+/PICK1+). The graphs show means ± SEM (one-way ANOVA; *** <span class="html-italic">p</span> &lt; 0.001, ** <span class="html-italic">p</span> &lt; 0.01, * <span class="html-italic">p</span> &lt; 0.05). The asterisks located directly above the bars signify that the difference applies to all time points within the group. The photomicrographs in the middle panel display the double-immunopositive reaction (yellow) for respective presynaptic (green) and postsynaptic (red) proteins, highlighting the time points for which the statistically significant differences were found in particular conditions. (<b>D</b>) Syp1+/PSD95+, (<b>E</b>) Pic+/Hom1+, (<b>F</b>) NRXN+/PICK1+. Scale bar, 10 µm. The right panel shows the averaged synapse density waveform, calculated across all protein pairs, for the different conditions. The experimental light conditions: light–dark 12 h:12 h (LD or LD12:12), constant darkness (DD), prolonged light LD 16 h:8 h (LD16 or LD 16:8), and constant light (LL). Time points are defined as follows: 0—ZT0/CT0: the beginning of the day/subjective day; 6—ZT6/CT6: the middle of the day/subjective day; 12—ZT12/CT12: the beginning of the night/subjective night; 18—ZT18/CT18: the middle of the night/subjective night.</p>
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<p>Participation of synaptic proteins in excitatory synapses. (<b>A</b>) Percentage participation of presynaptic and postsynaptic proteins in excitatory synapses. (<b>B</b>) Correlation between the area covered by a protein and the percentage participation of that protein in excitatory synapses for respective presynaptic (left column) and postsynaptic proteins (right column). Syp 1, Synaptophysin 1; Pic, Piccolo; NRXNs, Neurexin 1/2/3; Hom1, Homer 1; r, Pearson correlation coefficient.</p>
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14 pages, 5456 KiB  
Article
A Hybrid Photoplethysmography (PPG) Sensor System Design for Heart Rate Monitoring
by Farjana Akter Jhuma, Kentaro Harada, Muhamad Affiq Bin Misran, Hin-Wai Mo, Hiroshi Fujimoto and Reiji Hattori
Sensors 2024, 24(23), 7634; https://doi.org/10.3390/s24237634 - 29 Nov 2024
Viewed by 278
Abstract
A photoplethysmography (PPG) sensor is a cost-effective and efficacious way of measuring health conditions such as heart rate, oxygen saturation, and respiration rate. In this work, we present a hybrid PPG sensor system working in a reflective mode with an optoelectronic module, i.e., [...] Read more.
A photoplethysmography (PPG) sensor is a cost-effective and efficacious way of measuring health conditions such as heart rate, oxygen saturation, and respiration rate. In this work, we present a hybrid PPG sensor system working in a reflective mode with an optoelectronic module, i.e., the combination of an inorganic light-emitting diode (LED) and a circular-shaped organic photodetector (OPD) surrounding the LED for efficient light harvest followed by the proper driving circuit for accurate PPG signal acquisition. The performance of the hybrid sensor system was confirmed by the heart rate detection process from the PPG using fast Fourier transform analysis. The PPG signal obtained with a 50% LED duty cycle and 250 Hz sampling rate resulted in accurate heart rate monitoring with an acceptable range of error. The effects of the LED duty cycle and the LED luminous intensity were found to be crucial to the heart rate accuracy and to the power consumption, i.e., indispensable factors for the hybrid sensor. Full article
(This article belongs to the Section Biosensors)
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<p>(<b>a</b>) Schematic representation of the amplitude over time to explain PPG signal generation through tissues. (<b>b</b>) Schematic illustrations of transmissive mode PPG (LED and OPD are placed on the opposite side of the sensing location). (<b>c</b>) Schematic illustrations of reflective mode PPG (both LED and OPD are placed side by side on the sensing location).</p>
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<p>Block diagram of the proposed hybrid PPG sensor system.</p>
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<p>The layer structure of the OPD device.</p>
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<p>(<b>a</b>) Block diagram of the driving circuit containing TIA, filter, and amplifier. (<b>b</b>) Customized PCB including the driving circuit and LED.</p>
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<p>(<b>a</b>) FreeCAD design of the device holder. (<b>b</b>) Image of the 3D-printed device holder.</p>
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<p>The steps of arranging the PPG sensor system.</p>
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<p>(<b>a</b>) Schematic representation image of the OPD and actual image of prepared D1 device. (<b>b</b>–<b>d</b>) J–V characteristics of the prepared OPD devices D1, D2, and D3, respectively. The black and red solid lines indicate the dark and photocurrent densities, respectively. The photocurrent was obtained under a red LED illumination with a peak wavelength of 624 nm and a luminous intensity of 150.06 mcd. (<b>e</b>,<b>f</b>) On/off ratio of the OPD at different biasing voltages under the above-mentioned red and green LED illumination conditions.</p>
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<p>Obtained PPG signal with hybrid PPG sensor.</p>
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<p>Process flow chart of heart rate estimation.</p>
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<p>(<b>a</b>) PPG signal before filtering (some disturbance can be seen at the signal peak due to the disturbance introduced while measuring from fingertip). (<b>b</b>) PPG signal after applying a moving average filter with a window size 3 to smooth the signal. (<b>c</b>) PPG signal after moving average and Butterworth bandpass filter with a cut-off frequency of 0.5 Hz–16 Hz to remove unwanted frequency signals.</p>
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14 pages, 1213 KiB  
Review
The Role of Melatonin on Caprine (Capra hircus) Sperm Freezability: A Review
by Alberto Jorge Cardenas-Padilla, Francisco Jimenez-Trejo, Marco Cerbon and Alfredo Medrano
Antioxidants 2024, 13(12), 1466; https://doi.org/10.3390/antiox13121466 - 28 Nov 2024
Viewed by 420
Abstract
In mammals, the pineal hormone melatonin is the most powerful pacemaker of the master circadian clock and is responsible for reproduction in seasonal breeders. It is also well known that melatonin and its metabolites play antioxidant roles in many tissues, including reproductive cells. [...] Read more.
In mammals, the pineal hormone melatonin is the most powerful pacemaker of the master circadian clock and is responsible for reproduction in seasonal breeders. It is also well known that melatonin and its metabolites play antioxidant roles in many tissues, including reproductive cells. Melatonin synthesis and secretion from the pineal gland occurs during scotophase (the dark phase during a day–night cycle), while its inhibition is observed during photophase (period of light during a day–night cycle). Short-day breeders, such as goats, are stimulated to breed in a manner dependent on high endogenous levels of melatonin. This hormone can be synthesized in various extra-pineal tissues, such as retina, gastrointestinal tract, ovaries, and testis, with its main function being as a local antioxidant, given that melatonin and its metabolites are potent scavengers of reactive oxygen and nitrogen species. Moreover, it has been reported that some functions of melatonin can be exerted through plasma membrane and intracellular receptors expressed in the male reproductive system, including germ cells, immature and mature spermatozoa. It has been shown that melatonin may enhance gamete cryosurvival mainly by its addition into the media and/or in exogenous melatonin treatments in several species. In the present review, the physiological effects of endogenous melatonin in mammals are described, with a deeper focus on caprine reproduction. Additionally, results from recent investigations on the roles of exogenous melatonin aimed at improving the reproductive efficiency of goat bucks are discussed. There are contradictory findings and a limited amount of research available in the field of goat sperm cryopreservation associated with the use of melatonin. Understanding and improving goat reproduction and production is essential for many marginalized human populations around the world who directly depend on goats to maintain and improve their lifestyle. Full article
(This article belongs to the Special Issue Redox Regulation in Animal Reproduction)
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<p>Effects of temperature, osmotic, and oxidative stresses in spermatozoa during cryopreservation.</p>
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<p>Topographic localisation of MLT receptors in goat buck sperm plasma membrane and their possible effects on sperm physiology.</p>
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16 pages, 6174 KiB  
Article
Microstructure and Corrosion Behavior of PEO-Coated AA7075 Under Pulsed Unipolar Potential Control Mode
by Salvacion B. Orgen and Eden May B. Dela Pena
Coatings 2024, 14(12), 1498; https://doi.org/10.3390/coatings14121498 - 28 Nov 2024
Viewed by 383
Abstract
Plasma electrolytic oxidation (PEO) has emerged as a promising surface coating technique producing high-quality ceramic coating for light metals like Al, Mg, Ti, and their alloys. AA7075 is one of the commonly used Al alloys for aircraft structures, gears and shafts, and automotives [...] Read more.
Plasma electrolytic oxidation (PEO) has emerged as a promising surface coating technique producing high-quality ceramic coating for light metals like Al, Mg, Ti, and their alloys. AA7075 is one of the commonly used Al alloys for aircraft structures, gears and shafts, and automotives as it provides high yield and tensile strength. However, Al and its alloys have drawbacks that limit their further application. Thus, surface treatments are proposed to improve the metal and its alloy’s properties. In this study, the PEO of AA7075 was carried out with an AC power source under a pulsed unipolar potentiostatic mode at varying voltages of 425 and 450 V in 1000 Hz and at 80% duty cycles of 30 m. The effect of varying voltages on the morphology, coating thickness, and corrosion resistance of the PEO-coated samples was investigated. Surface morphology, elemental distribution, and phase composition were characterized using SEM, EDX, and XRD. A porous structure with a pancake-like shape, a crater, and nodular structures were observed with coating thickness ranges from 12.1 to 55.3 ± 4.67 µm. Al, α-alumina, and γ-alumina were detected in all surface coatings. The PEO-coated sample at 450 V exhibited higher corrosion resistance evaluated via potentiodynamic polarization and EIS. Full article
(This article belongs to the Section Ceramic Coatings and Engineering Technology)
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<p>Schematic of (<b>a</b>) pulsed bipolar and (<b>b</b>) pulsed unipolar waveforms in PEO process.</p>
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<p>SEM image of PEO-coated AA7075 at discharge potentials of (<b>a</b>,<b>b</b>) 425 V and (<b>c</b>,<b>d</b>) 450 V.</p>
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<p>Pore density and average pore diameter of PEO-coated AA7075 at 425 and 450 V.</p>
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<p>Histogram of pore size distribution PEO coating at 425 V and 450 V.</p>
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<p>Cross-section of PEO-coated AA7075 at discharge potentials of (<b>a</b>) 425 and (<b>b</b>) 450 V. Inset figures of PEO coating (dotted line).</p>
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<p>EDX mapping of PEO-coated AA7075 at (<b>a</b>) 425 and (<b>b</b>) 450 V.</p>
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<p>XRD of PEO-coated AA7075 at 425 V and 450 V.</p>
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<p>Potentiodynamic polarization of bare and PEO-coated AA7075.</p>
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<p>EIS measurement of bare and PEO-coated AA7075 samples in 3.5 wt% NaCl (<b>a</b>) Nyquist plot, (<b>b</b>) Bode-phase and (<b>c</b>) Bode-modulus plots.</p>
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<p>Proposed equivalent electrical circuits for the (<b>a</b>) uncoated AA7075 and PEO-coated samples at (<b>b</b>) 425 V and (<b>c</b>) 450 V.</p>
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<p>Photographs of the specimen during PEO coating after (<b>a</b>) 1–3 m and (<b>b</b>) 30 m.</p>
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18 pages, 3034 KiB  
Article
Water Conflicts and Socioterritorial Dynamics: The Hydrosocial Cycle After the São Francisco River Transposition Project in the Northeast of Brazil
by Jaqueline Guimarães Santos and Antonio A. R. Ioris
Land 2024, 13(12), 2032; https://doi.org/10.3390/land13122032 - 28 Nov 2024
Viewed by 325
Abstract
The implementation of large-scale water infrastructure projects, such as the São Francisco River Integration Project with the Northeastern Hydrographic Basins (PISF), causes territorial transformations of great proportions, constituting a new hydrosocial cycle, resulting in hydrosocial territories in which not only water but also [...] Read more.
The implementation of large-scale water infrastructure projects, such as the São Francisco River Integration Project with the Northeastern Hydrographic Basins (PISF), causes territorial transformations of great proportions, constituting a new hydrosocial cycle, resulting in hydrosocial territories in which not only water but also power relations and socioeconomic and physical resources circulate. In this sense, this article aims to historically analyze the interconnection between water and land issues in the Brazilian Northeast, shedding light on the territorial dynamics produced by the new hydrosocial cycle constituted by the implementation of the largest water project in the country. To this end, we interviewed actors from different groups and in various locations, conducted on-site field research, and collected and systematically analyzed important documents from the institutional websites of the Brazilian government. The main results of the research indicate that since the implementation of the PISF, a context of deep and historical political and socioeconomic inequalities in hydrosocial territories has been impacted, which has deepened conflicts over land and water and increased socio-spatial injustices. Contrary to most available approaches, which consider water scarcity a ‘natural’ (hydrological) problem that requires technical and administrative solutions, the water problem reflects the profound asymmetry of power consolidated over time and the strength of the reactionary agrarian sectors that control the State apparatus. Full article
(This article belongs to the Section Water, Energy, Land and Food (WELF) Nexus)
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<p>Hydrosocial cycle. Source: [<a href="#B28-land-13-02032" class="html-bibr">28</a>] (p. 176).</p>
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<p>Locations of rural communities visited in the field research. Source: Prepared based on data from the IBGE cartographic database (2020).</p>
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<p>Periodization of droughts and main actions and projects around the water issue in the Brazilian Northeast. Source: [<a href="#B12-land-13-02032" class="html-bibr">12</a>].</p>
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<p>PISF hydrosocial territories in Pernambuco, Brazil. Source: Own collection.</p>
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<p>Hydrosocial cycle and multiple relationships.</p>
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21 pages, 5787 KiB  
Article
Only Infant MLL-Rearranged Leukemia Is Susceptible to an Inhibition of Polo-like Kinase 1 (PLK-1) by Volasertib
by Jacqueline Fischer, Estelle Erkner, Pia Radszuweit, Thomas Hentrich, Hildegard Keppeler, Fulya Korkmaz, Julia Schulze-Hentrich, Rahel Fitzel, Claudia Lengerke, Dominik Schneidawind and Corina Schneidawind
Int. J. Mol. Sci. 2024, 25(23), 12760; https://doi.org/10.3390/ijms252312760 - 27 Nov 2024
Viewed by 318
Abstract
MLL-rearranged (MLLr) leukemia is characterized by a poor prognosis. Depending on the cell of origin, it differs in the aggressiveness and therapy response. For instance, in adults, volasertib blocking Polo-like kinase 1 (PLK-1) exhibited limited success. Otherwise, PLK-1 characterizes an [...] Read more.
MLL-rearranged (MLLr) leukemia is characterized by a poor prognosis. Depending on the cell of origin, it differs in the aggressiveness and therapy response. For instance, in adults, volasertib blocking Polo-like kinase 1 (PLK-1) exhibited limited success. Otherwise, PLK-1 characterizes an infant MLLr signature, indicating potential sensitivity. By using our CRISPR/Cas9 MLLr model in CD34+ cells from human cord blood (huCB) and bone marrow (huBM) mimicking the infant and adult patient diseases, we were able to shed light on this phenomenon. The PLK-1 mRNA level was significantly increased in our huCB compared to the huBM model, which was underpinned by analyzing infant and adult MLLr leukemia patients. Importantly, the expression levels correlated with a functional response. Volasertib induced a significant dose-dependent decrease in proliferation and cell cycle arrest, most pronounced in the infant model. Mechanistically, upon volasertib treatment, we uncovered negative feedback only in the huBM model by compensatory upregulation of PLK-1 and related genes like AURKA involved in mitosis. Importantly, the poor response could be overcome by a combinatorial strategy with alisertib, an Aurora kinase A inhibitor. Our study emphasizes the importance of considering the cell of origin in therapeutic decision-making and provides the rationale for evaluating volasertib and alisertib in MLLr leukemia. Full article
(This article belongs to the Special Issue Hallmarks of Cancer: Emerging Insights and Innovations)
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<p><b>Revealing PLK-1 as a potential promising target in infant MLL-AF9 leukemia.</b> (<b>A</b>) CD34+ HSPCs were isolated from huCB and huBM via Ficoll separation and magnetic cell separation and cultured for 48 h. Thereafter, <span class="html-italic">t(9;11)</span> was induced in cultured HSPCs using the CRISPR/Cas9 system. (<b>B</b>) RNA sequencing of human CRISPR/Cas9 <span class="html-italic">MLL-AF9</span> cells derived from huCB (<span class="html-italic">n</span> = 5) and huBM (<span class="html-italic">n</span> = 5) compared with the respective control cells (huCB/huBM-derived CD34+, <span class="html-italic">n</span> = 4). Student’s <span class="html-italic">t</span>-test. * <span class="html-italic">p</span> &lt; 0.05. ns: not significant <span class="html-italic">p</span> &gt; 0.05. (<b>C</b>) Fold change of <span class="html-italic">PLK-1</span> overexpression in huCB <span class="html-italic">MLL-AF4</span> and <span class="html-italic">MLL-AF9</span> cells (<span class="html-italic">n</span> = 5/<span class="html-italic">n</span> = 5); huBM <span class="html-italic">MLL</span>r cells (<span class="html-italic">n</span> = 5); cell lines THP-1, NOMO-1, SEM, RS4;11, KOPN8, and SKM1 (all <span class="html-italic">n</span> = 3); and infant and adult <span class="html-italic">MLL</span>r leukemia patient samples (<span class="html-italic">n</span> = 3/<span class="html-italic">n</span> = 3) compared to CD34+ huCB/BM control cells (ctrl, <span class="html-italic">n</span> = 4), measured by RT-qPCR. One-way ANOVA. * <span class="html-italic">p</span> &lt; 0.05. ns: not significant <span class="html-italic">p</span> &gt; 0.05. Overview of <span class="html-italic">MLL</span> translocation in the cell lines used. (<b>D</b>) Kaplan–Meier survival curve (<a href="http://www.kmplot.com" target="_blank">www.kmplot.com</a>, 26 August 2024). Higher <span class="html-italic">PLK-1</span> expression levels in AML patients show a trend to worse survival rates. Logrank. <span class="html-italic">p</span> = 0.083. Median survival rates [months]: low <span class="html-italic">PLK-1</span> level 16.4, high <span class="html-italic">PLK-1</span> level 12.1.</p>
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<p><b><span class="html-italic">MLL</span>r leukemia cells derived from huCB are more susceptible to a PLK-1 inhibition than those from huBM.</b> (<b>A</b>) Cell counts were assessed following treatment with volasertib (50 nM), vehicle control (DMSO), or no treatment (as a baseline control) in huCB and huBM CRISPR/Cas9 <span class="html-italic">MLL</span>r cells (<span class="html-italic">n</span> = 3 each). Relative cell counts were determined using a Neubauer counting chamber after Trypan blue staining and normalized to the vehicle control (DMSO). Right: Significant difference of proliferation between huCB and huBM CRISPR/Cas9 <span class="html-italic">MLL</span>r cells at 72 h after 50 nM volasertib treatment. One-way ANOVA. * <span class="html-italic">p</span> &lt; 0.05. (<b>B</b>) huCB and huBM CRISPR/Cas9 <span class="html-italic">MLL</span>r (<span class="html-italic">n</span> = 3/<span class="html-italic">n</span> = 3) and CD34+ huCB/BM control cells (ctrl, <span class="html-italic">n</span> = 4) were treated with increasing concentrations of volasertib or vehicle control (DMSO) for 72 h. Relative cell count was determined by counting cells in a Neubauer counting chamber after Trypan blue staining, normalized to vehicle control (DMSO). IC50 values: huBM <span class="html-italic">MLL</span>r 43.5 nM, huCM <span class="html-italic">MLL</span>r 17.9 nM, ctrl 35.1 nM. IC50 values of the dose-dependent curves were interpolated from a four-parameter logistic model. (<b>C</b>) Significant difference between IC50 values of huBM <span class="html-italic">MLL</span>r and huCB <span class="html-italic">MLL</span>r cells compared to ctrl and each other. One-way ANOVA. * <span class="html-italic">p</span> &lt; 0.05. ns: not significant <span class="html-italic">p</span> &gt; 0.05. (<b>D</b>) Representative flow cytometric histograms of Annexin V/PI staining to determine the apoptotic effect of 72 h volasertib treatment (DMSO, 50 nM, 100 nM) on huCB and huBM CRISPR/Cas9 <span class="html-italic">MLL</span>r (<span class="html-italic">n</span> = 3/<span class="html-italic">n</span> = 3) and CD34+ huCB/BM control cells (ctrl, <span class="html-italic">n</span> = 4), measured by flow cytometry. On the right, summarized fractions normalized to their own vehicle control (DMSO). One-way ANOVA. * <span class="html-italic">p</span> &lt; 0.05. (<b>E</b>) Left: THP-1 and NOMO-1 (<span class="html-italic">n</span> = 3/<span class="html-italic">n</span> = 3) cells were treated with increasing concentrations of volasertib or vehicle control (DMSO) for 72 h. Relative cell count was determined by counting cells in a Neubauer counting chamber after Trypan blue staining and normalized to vehicle control (DMSO). IC50 values: THP-1 8.0 nM; NOMO-1 could not be determined, as 50% cell death was not achieved (n.c.). IC50 values of the dose-dependent curves were interpolated from a four-parameter logistic model. Significant difference between endpoints (200 nM volasertib treatment) of NOMO-1 and THP-1. One-way ANOVA. * <span class="html-italic">p</span> &lt; 0.05. Right: Representative flow cytometric histograms and summarized distribution of Annexin V/PI staining to determine the apoptotic effect of volasertib treatment (DMSO, 50 nM, 100 nM, 72 h incubation) on NOMO-1 and THP-1 (<span class="html-italic">n</span> = 3/<span class="html-italic">n</span> = 3) cells measured by flow cytometry. On the right, summarized fractions normalized to their own vehicle control (DMSO). One-way ANOVA. * <span class="html-italic">p</span> &lt; 0.05.</p>
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<p><b>Inhibition of PLK-1 leads to reduced viability and mitotic arrest.</b> (<b>A</b>) The 72 h volasertib treatment (DMSO vehicle control, 50 nM, 100 nM) on huBM and huCB CRISPR/Cas9 <span class="html-italic">MLL</span>r cells (<span class="html-italic">n</span> = 3/<span class="html-italic">n</span> = 3) decreased cell viability, measured by AlamarBlue viability assay. Comparison of the reduction in cell viability after treatment with 100 nM volasertib between huBM and huCB CRISPR/Cas9 <span class="html-italic">MLL</span>r cells (<span class="html-italic">n</span> = 3/<span class="html-italic">n</span> = 3) and CD34+ huCB/BM control cells (ctrl, <span class="html-italic">n</span> = 4). One-way ANOVA. * <span class="html-italic">p</span> &lt; 0.05. ns: not significant <span class="html-italic">p</span> &gt; 0.05. (<b>B</b>) Representative (left) and pooled (right) data of BrdU cell cycle analysis of huBM and huCB CRISPR/Cas9 <span class="html-italic">MLL</span>r cells (<span class="html-italic">n</span> = 3/<span class="html-italic">n</span> = 3), CD34+ huCB control cells (ctrl, <span class="html-italic">n</span> = 3), and THP-1, NOMO-1 (<span class="html-italic">n</span> = 3/<span class="html-italic">n</span> = 3) after volasertib treatment for 48 h (DMSO vehicle control, 50 nM, 100 nM). This shows a significant increase in G2/M-phase and a decrease in S-phase in CB <span class="html-italic">MLL</span>r cells. Normalized to respective vehicle control (DMSO). One-way ANOVA. * <span class="html-italic">p</span> &lt; 0.05. ns: not significant <span class="html-italic">p</span> &gt; 0.05. (<b>C</b>) Images show representative morphologies of huBM and huCB CRISPR/Cas9 <span class="html-italic">MLL</span>r cells after volasertib treatment (DMSO vehicle control, 50 nM, 100 nM). Black arrows point at mitotic figures; cells arrested in M-phase. Pappenheim staining.</p>
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<p><b>Inhibition of PLK-1 leads to reduced viability and mitotic arrest.</b> (<b>A</b>) The 72 h volasertib treatment (DMSO vehicle control, 50 nM, 100 nM) on huBM and huCB CRISPR/Cas9 <span class="html-italic">MLL</span>r cells (<span class="html-italic">n</span> = 3/<span class="html-italic">n</span> = 3) decreased cell viability, measured by AlamarBlue viability assay. Comparison of the reduction in cell viability after treatment with 100 nM volasertib between huBM and huCB CRISPR/Cas9 <span class="html-italic">MLL</span>r cells (<span class="html-italic">n</span> = 3/<span class="html-italic">n</span> = 3) and CD34+ huCB/BM control cells (ctrl, <span class="html-italic">n</span> = 4). One-way ANOVA. * <span class="html-italic">p</span> &lt; 0.05. ns: not significant <span class="html-italic">p</span> &gt; 0.05. (<b>B</b>) Representative (left) and pooled (right) data of BrdU cell cycle analysis of huBM and huCB CRISPR/Cas9 <span class="html-italic">MLL</span>r cells (<span class="html-italic">n</span> = 3/<span class="html-italic">n</span> = 3), CD34+ huCB control cells (ctrl, <span class="html-italic">n</span> = 3), and THP-1, NOMO-1 (<span class="html-italic">n</span> = 3/<span class="html-italic">n</span> = 3) after volasertib treatment for 48 h (DMSO vehicle control, 50 nM, 100 nM). This shows a significant increase in G2/M-phase and a decrease in S-phase in CB <span class="html-italic">MLL</span>r cells. Normalized to respective vehicle control (DMSO). One-way ANOVA. * <span class="html-italic">p</span> &lt; 0.05. ns: not significant <span class="html-italic">p</span> &gt; 0.05. (<b>C</b>) Images show representative morphologies of huBM and huCB CRISPR/Cas9 <span class="html-italic">MLL</span>r cells after volasertib treatment (DMSO vehicle control, 50 nM, 100 nM). Black arrows point at mitotic figures; cells arrested in M-phase. Pappenheim staining.</p>
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<p><b>Transcriptomic analysis</b> revealed a compensatory PLK-1 feedback mechanism only in adult <span class="html-italic">MLL</span>r cells upon volasertib treatment. huBM and huCB CRISPR/Cas9 <span class="html-italic">MLL</span>r cells (<span class="html-italic">n</span> = 4/<span class="html-italic">n</span> = 3) were treated with 50 nM volasertib or vehicle control (DMSO) for 72 h and used for RNA-seq. (<b>A</b>) Analysis revealed in huBM <span class="html-italic">MLL</span>r cells 728 differentially expressed genes (DEGs) and 151 DEGs in huCB <span class="html-italic">MLL</span>r cells after volasertib treatment. Volcano plot of huBM and huCB CRISPR/Cas9 <span class="html-italic">MLL</span>r cells after volasertib treatment highlighting downregulated (blue) and upregulated (red) DEGs. Dotted lines indicate significant thresholds (pFDR ≤ 0.05, |log2(fold-change)| ≥  0.5). (<b>B</b>) log2(fold-change) of DEGs are shown for <span class="html-italic">PLK-1</span> and associated genes, normalized to own vehicle control (DMSO). Black * p_adjust value, white * p_nominal value. huBM <span class="html-italic">MLL</span>r cells show an upregulation of PLK-1-related gene pattern, whereas the pattern of huCB <span class="html-italic">MLL</span>r is downregulated. (<b>C</b>) Interactome of significant altered normalized Reads per Kilobase Millions (nRPKMs) around <span class="html-italic">PLK-1</span> in huBM and huCB CRISPR/Cas9 <span class="html-italic">MLL</span>r cells reveals potential feedback mechanism. Upregulation (red), downregulation (blue). Additional RNA-seq analysis regarding phosphorylation activity and targets around <span class="html-italic">PLK-1</span> are highlighted with red arrows. (<b>D</b>) Fold change of <span class="html-italic">PLK-1</span>, <span class="html-italic">BORA</span>, <span class="html-italic">AURKA,</span> and <span class="html-italic">FOXM1</span> in huBM and huCB CRISPR/Cas9 <span class="html-italic">MLL</span>r cells (<span class="html-italic">n</span> = 4/<span class="html-italic">n</span> = 3), as well as NOMO-1 and THP-1 cells (<span class="html-italic">n</span> = 3/<span class="html-italic">n</span> = 3), after 72 h 50 nM volasertib treatment compared to vehicle control (DMSO) measured by RT-qPCR. One-way ANOVA. * <span class="html-italic">p</span> &lt; 0.05.</p>
Full article ">Figure 4 Cont.
<p><b>Transcriptomic analysis</b> revealed a compensatory PLK-1 feedback mechanism only in adult <span class="html-italic">MLL</span>r cells upon volasertib treatment. huBM and huCB CRISPR/Cas9 <span class="html-italic">MLL</span>r cells (<span class="html-italic">n</span> = 4/<span class="html-italic">n</span> = 3) were treated with 50 nM volasertib or vehicle control (DMSO) for 72 h and used for RNA-seq. (<b>A</b>) Analysis revealed in huBM <span class="html-italic">MLL</span>r cells 728 differentially expressed genes (DEGs) and 151 DEGs in huCB <span class="html-italic">MLL</span>r cells after volasertib treatment. Volcano plot of huBM and huCB CRISPR/Cas9 <span class="html-italic">MLL</span>r cells after volasertib treatment highlighting downregulated (blue) and upregulated (red) DEGs. Dotted lines indicate significant thresholds (pFDR ≤ 0.05, |log2(fold-change)| ≥  0.5). (<b>B</b>) log2(fold-change) of DEGs are shown for <span class="html-italic">PLK-1</span> and associated genes, normalized to own vehicle control (DMSO). Black * p_adjust value, white * p_nominal value. huBM <span class="html-italic">MLL</span>r cells show an upregulation of PLK-1-related gene pattern, whereas the pattern of huCB <span class="html-italic">MLL</span>r is downregulated. (<b>C</b>) Interactome of significant altered normalized Reads per Kilobase Millions (nRPKMs) around <span class="html-italic">PLK-1</span> in huBM and huCB CRISPR/Cas9 <span class="html-italic">MLL</span>r cells reveals potential feedback mechanism. Upregulation (red), downregulation (blue). Additional RNA-seq analysis regarding phosphorylation activity and targets around <span class="html-italic">PLK-1</span> are highlighted with red arrows. (<b>D</b>) Fold change of <span class="html-italic">PLK-1</span>, <span class="html-italic">BORA</span>, <span class="html-italic">AURKA,</span> and <span class="html-italic">FOXM1</span> in huBM and huCB CRISPR/Cas9 <span class="html-italic">MLL</span>r cells (<span class="html-italic">n</span> = 4/<span class="html-italic">n</span> = 3), as well as NOMO-1 and THP-1 cells (<span class="html-italic">n</span> = 3/<span class="html-italic">n</span> = 3), after 72 h 50 nM volasertib treatment compared to vehicle control (DMSO) measured by RT-qPCR. One-way ANOVA. * <span class="html-italic">p</span> &lt; 0.05.</p>
Full article ">Figure 5
<p><b>Combined treatment of volasertib and alisertib in <span class="html-italic">MLL</span>r cells.</b> (<b>A</b>) huCB <span class="html-italic">MLL</span>r and huBM <span class="html-italic">MLL</span>r (<span class="html-italic">n</span> = 3/<span class="html-italic">n</span> = 3) cells were treated for 72 h with increasing concentrations of volasertib alone, alisertib alone, or in combination. The percentage of viable cells (Annexin V-, PI-) was determined by flow cytometry. In the isobologram, IC50 values were mapped, and the Chou–Talalay method was used to measure the CI for the identification of synergistic effects. CI: huBM <span class="html-italic">MLL</span>r 0.25, huCB <span class="html-italic">MLL</span>r 1.52. (<b>B</b>) Representative (<b>left</b>) and pooled (<b>right</b>) data of BrdU cell cycle analysis of huBM <span class="html-italic">MLL</span>r cells (<span class="html-italic">n</span> = 3) after combinatorial treatment with volasertib and alisertib for 72 h (DMSO vehicle control, 25 nM V + 5 µM A, 100 nM V + 50 µM A). This shows a significant increase in G2/M-phase and apoptotic cells and a decrease in S-phase. Normalized to respective vehicle control (DMSO). One-way ANOVA. * <span class="html-italic">p</span> &lt; 0.05.</p>
Full article ">Figure 5 Cont.
<p><b>Combined treatment of volasertib and alisertib in <span class="html-italic">MLL</span>r cells.</b> (<b>A</b>) huCB <span class="html-italic">MLL</span>r and huBM <span class="html-italic">MLL</span>r (<span class="html-italic">n</span> = 3/<span class="html-italic">n</span> = 3) cells were treated for 72 h with increasing concentrations of volasertib alone, alisertib alone, or in combination. The percentage of viable cells (Annexin V-, PI-) was determined by flow cytometry. In the isobologram, IC50 values were mapped, and the Chou–Talalay method was used to measure the CI for the identification of synergistic effects. CI: huBM <span class="html-italic">MLL</span>r 0.25, huCB <span class="html-italic">MLL</span>r 1.52. (<b>B</b>) Representative (<b>left</b>) and pooled (<b>right</b>) data of BrdU cell cycle analysis of huBM <span class="html-italic">MLL</span>r cells (<span class="html-italic">n</span> = 3) after combinatorial treatment with volasertib and alisertib for 72 h (DMSO vehicle control, 25 nM V + 5 µM A, 100 nM V + 50 µM A). This shows a significant increase in G2/M-phase and apoptotic cells and a decrease in S-phase. Normalized to respective vehicle control (DMSO). One-way ANOVA. * <span class="html-italic">p</span> &lt; 0.05.</p>
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