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Search Results (1,357)

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12 pages, 22317 KiB  
Article
Biomimetic Cooling: Functionalizing Biodegradable Chitosan Films with Saharan Silver Ant Microstructures
by Markus Zimmerl, Richard W. van Nieuwenhoven, Karin Whitmore, Wilfried Vetter and Ille C. Gebeshuber
Biomimetics 2024, 9(10), 630; https://doi.org/10.3390/biomimetics9100630 - 17 Oct 2024
Viewed by 165
Abstract
The increasing occurrence of hot summer days causes stress to both humans and animals, particularly in urban areas where temperatures can remain high, even at night. Living nature offers potential solutions that require minimal energy and material costs. For instance, the Saharan silver [...] Read more.
The increasing occurrence of hot summer days causes stress to both humans and animals, particularly in urban areas where temperatures can remain high, even at night. Living nature offers potential solutions that require minimal energy and material costs. For instance, the Saharan silver ant (Cataglyphis bombycina) can endure the desert heat by means of passive radiative cooling induced by its triangular hairs. The objective of this study is to transfer the passive radiative cooling properties of the micro- and nanostructured chitin hairs of the silver ant body to technically usable, biodegradable and bio-based materials. The potential large-scale transfer of radiative cooling properties, for example, onto building exteriors such as house facades, could decrease the need for conventional cooling and, therefore, lower the energy demand. Chitosan, a chemically altered form of chitin, has a range of medical uses but can also be processed into a paper-like film. The procedure consists of dissolving chitosan in diluted acetic acid and uniformly distributing it on a flat surface. A functional structure can then be imprinted onto this film while it is drying. This study reports the successful transfer of the microstructure-based structural colors of a compact disc (CD) onto the film. Similarly, a polyvinyl siloxane imprint of the silver ant body shall make it possible to transfer cooling functionality to technically relevant surfaces. FTIR spectroscopy measurements of the reflectance of flat and structured chitosan films allow for a qualitative assessment of the infrared emissivity. A minor decrease in reflectance in a relevant wavelength range gives an indication that it is feasible to increase the emissivity and, therefore, decrease the surface temperature purely through surface-induced functionalities. Full article
(This article belongs to the Special Issue The Latest Progress in Bionics Research)
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Graphical abstract

Graphical abstract
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<p>Incoming and outgoing radiation energy intensity and the absorption spectrum of the atmosphere. The bulk of the outgoing energy lies within the atmospheric window from 8 to 13 µm [<a href="#B17-biomimetics-09-00630" class="html-bibr">17</a>].</p>
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<p>(<b>a</b>) SEM micrograph of cross-sections cut with a focused ion beam (FIB) through triangular chitin hairs of a Sahara silver ant gaster (hind part). Scale bar—2 µm. (<b>b</b>) Illustration of the triangular cross-section of a silver ant hair. Incoming solar radiation undergoes Mie scattering at the small indentations of the top sides. The light that enters the silver ant hair can be reflected on the bottom side when the conditions for total reflection are met (incidence angle and difference in refractive index between silver ant hair and air gap) [<a href="#B14-biomimetics-09-00630" class="html-bibr">14</a>]. Scale bar—1 µm.</p>
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<p>(<b>a</b>) SEM sample holder with exposed and unexposed shrimp shell sample, as well as silver ant gaster (rear segment of the silver ant). Scale bar—1 cm. (<b>b</b>) Climate chamber cycles (programmed).</p>
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<p>Process of creating a copy of the silver ant surface structure in chitosan with the help of a PVS stamp.</p>
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<p>(<b>a</b>) Confocal image of scratched shrimp shell before and after exposure in the climate chamber. Scale bar—200 µm. (<b>b</b>) Chitosan film with iridescent microstructures transferred from a CD. Scale bar—0.5 cm.</p>
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<p>Confocal images of (<b>a</b>) an ’unstructured’ area of the PVS stamp, which shows the structure of the cardboard that surrounded the silver ant gaster. (<b>b</b>) The PVS–cardboard structure transferred onto chitosan. (<b>c</b>) A structured area of the PVS stamp structured with an ant gaster. (<b>d</b>) The PVS–ant structure transferred onto chitosan. Scale bars—200 µm.</p>
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<p>(<b>a</b>) Average reflectance (* in relation to a reference gold mirror) of structured and unstructured chitosan films. The structured areas in both samples exhibit a slightly higher reflectance for wavelengths greater than 6 µm. However, this difference is less than the calculated standard deviation. Below 6 µm, the two samples feature inconsistent differences in reflectance. (<b>b</b>) Zoom into the respective region of the atmospheric window in (<b>a</b>).</p>
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24 pages, 12513 KiB  
Article
Genome-Wide Identification of the Maize Chitinase Gene Family and Analysis of Its Response to Biotic and Abiotic Stresses
by Tonghan Wang, Changjin Wang, Yang Liu, Kunliang Zou, Minghui Guan, Yutong Wu, Shutong Yue, Ying Hu, Haibing Yu, Kaijing Zhang, Degong Wu and Junli Du
Genes 2024, 15(10), 1327; https://doi.org/10.3390/genes15101327 (registering DOI) - 15 Oct 2024
Viewed by 378
Abstract
Background/Objectives: Chitinases, enzymes belonging to the glycoside hydrolase family, play a crucial role in plant growth and stress response by hydrolyzing chitin, a natural polymer found in fungal cell walls. This study aimed to identify and analyze the maize chitinase gene family, assessing [...] Read more.
Background/Objectives: Chitinases, enzymes belonging to the glycoside hydrolase family, play a crucial role in plant growth and stress response by hydrolyzing chitin, a natural polymer found in fungal cell walls. This study aimed to identify and analyze the maize chitinase gene family, assessing their response to various biotic and abiotic stresses to understand their potential role in plant defense mechanisms and stress tolerance. Methods: We employed bioinformatics tools to identify 43 chitinase genes in the maize B73_V5 genome. These genes were characterized for their chromosomal positions, gene and protein structures, phylogenetic relationships, functional enrichment, and collinearity. Based on previous RNA-seq data, the analysis assessed the expression patterns of these genes at different developmental stages and under multiple stress conditions. Results: The identified chitinase genes were unevenly distributed across maize chromosomes with a history of tandem duplications contributing to their divergence. The ZmChi protein family was predominantly hydrophilic and localized mainly in chloroplasts. Expression analysis revealed that certain chitinase genes were highly expressed at specific developmental stages and in response to various stresses, with ZmChi31 showing significant responsiveness to 11 different abiotic and biotic stresses. Conclusions: This study provides new insights into the role of chitinase genes in maize stress response, establishing a theoretical framework for exploring the molecular basis of maize stress tolerance. The identification of stress-responsive chitinase genes, particularly ZmChi31, offers potential candidates for further study in enhancing maize resistance to environmental challenges. Full article
(This article belongs to the Special Issue Maize Molecular Genetics and Functional Genomics in 2024)
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Figure 1
<p>A heat map depicting the subcellular localization of all ZmChi genes within plant cells, including the plasmid, endoplasmic reticulum, Golgi apparatus, vesicles, mitochondria, chloroplasts, nucleus, cytoplasm, ectoplasm, and peroxisomes. Blue indicates the lack of the gene in that area, gray indicates a small functional presence, and red indicates the highest functional importance of the gene in that specific region. Note: nucl: nucleus; golg: Golgi; vacu: vesicle; cyto: cytoplasm; plas: plasmid; mito: mitochondrion; chlo: chloroplast; extr: outer matrix; E.R.: endoplasmic reticulum; pero: peroxisome.</p>
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<p>Chromosome distribution of chitinase genes in maize. Note: Genes are color-coded based on duplication type: green for tandem duplicates, blue for segmental duplicates, and red for those that are both tandem and segmental duplicates.</p>
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<p>Phylogenetic analysis of rice, maize, and Arabidopsis chitinase proteins. GH18 comprises family members from groups III and V; GH19 encompasses family members from groups I, II, and IV; and GH20 consists of family members from group VIII.</p>
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<p>A visual representation showing the preserved pattern of protein motifs and the arrangement of exons and introns in the ZmChi genes of maize. GH18 comprises family members from groups III and V; GH19 encompasses family members from groups I, II, and IV; and GH20 consists of family members from group VIII.</p>
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<p>Investigation of cis-regulatory elements within the promoter regions of the maize chitinase gene family: (<b>A</b>) Examination of the prevalence of diverse cis-regulatory elements across these promoters. (<b>B</b>) Assessment of the frequency of various cis-regulatory element types within these promoter sequences.</p>
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<p>Fold enrichment diagram highlighting the shared functions of ZmChi genes: (<b>A</b>) GO terms associated with biological processes; (<b>B</b>) GO terms associated with cellular components; (<b>C</b>) GO terms associated with molecular functions. Note: Red dot plots indicate a greater number of genes participating in each respective process, and blue dots of smaller size indicate fewer genes.</p>
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<p>Duplication and synteny of maize chitinase genes. (<b>A</b>) The diagram illustrates the homologous relationships of maize chitinase genes with counterparts in <span class="html-italic">Arabidopsis thaliana</span> and rice, where green lines denote homologous gene pairs between maize and rice, and blue lines represent those between maize and <span class="html-italic">Arabidopsis thaliana</span>. (<b>B</b>) The horizontal axis represents tandem repeats, segmental repeats, and maize repeats with <span class="html-italic">Arabidopsis thaliana</span> (Zm-At) and rice (Zm-Os). The vertical axis denotes the corresponding numerical values.</p>
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<p>A heatmap shows the heterogeneity in expression patterns of chitinase genes across maize tissues. 24H_Germinating Seed: Seed 24 h after germination begins; 6DAS_GH_Primary Root: Root at 6 days after seed germination; V3_Stem and SAM: Stem and Shoot Apical Meristem (SAM) at the V3 corn growth stage; V5_Tip of stage-2 Leaf: Tip of the stage-2 leaf at the V5 corn growth stage; V9_Immature Leaves: Immature leaves at the V9 corn growth stage; V9_Thirteenth Leaf: Thirteenth leaf at the V9 corn growth stage; V9_Eleventh Leaf: Eleventh leaf at the V9 corn growth stage; V9_Eighth Leaf: Eighth leaf at the V9 corn growth stage; VT_Thirteenth Leaf: Thirteenth leaf at the Tasseling (VT) corn growth stage; R2_Thirteenth Leaf: Thirteenth leaf at the R2 reproductive stage of corn; 10DAP_Whole seed: Whole seed at 10 days after pollination; 12DAP_Whole seed: Whole seed at 12 days after pollination; 14DAP_Whole seed: Whole seed at 14 days after pollination; 16DAP_Whole seed: Whole seed at 16 days after pollination; 12DAP_Endopsperm: Endosperm at 12 days after pollination; 14DAP_Endopsperm: Endosperm at 14 days after pollination; 16DAP_Endopsperm: Endosperm at 16 days after pollination; 16DAP_Embryo: Embryo at 16 days after pollination. Note: The boxed data represent raw FPKM values.</p>
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<p>A heatmap displaying maize chitinase gene expression under different abiotic stress conditions. (<b>A</b>) The expression patterns of maize chitinase genes were studied under various temperature stress conditions, including NT: normal (25 °C), ELT: extremely low (4 °C), MLT: medium-low (10 °C), HT: high (37 °C), LT: low (16 °C), MHT: medium-high (42 °C), and EHT: extremely high (48 °C) temperatures. (<b>B</b>) We analyzed the temporal expression patterns of maize chitinase genes under flooding stress at various intervals: 0 h (ws-0 h), 2 h (ws-2 h), 4 h (ws-4 h), 6 h (ws-6 h), 8 h (ws-8 h), 10 h (ws-10 h), and 12 h (ws-12 h). (<b>C</b>) Analysis of the expression of maize chitinase genes under drought stress conditions revealed patterns using different sample types: YZXC for moisture-treated samples from the drought-intolerant hybrid ZX978, YZXD for drought-treated samples from the drought-intolerant hybrid ZX978, YHNC for moisture-treated samples from the drought-tolerant hybrid ND476, and YHND for drought-treated samples from the drought-tolerant hybrid ND476. (<b>D</b>) The response of maize chitinase genes to salt stress was investigated by subjecting the salt-tolerant inbred line L87 (T-salt) and the salt-sensitive inbred line L29 (S-salt) to a 220 mM NaCl treatment. Concurrently, control groups T-CT and S-CT for L87 and L29, respectively, were treated with 0 mM NaCl. (<b>E</b>) Under varying light conditions, the maize chitinase gene family was evaluated. Here, SNL represented seedlings under standard light conditions; SDT indicated seedlings subjected to four days of darkness; and SRNL denoted seedlings that were first treated with four days of darkness followed by four days of normal light. Note: Each chart depicts the original FPKM value in the left box and the value of log2(multiple changes) in the right box, grey (no significant differences), highlighted in red (up-regulated), and green (down-regulated), |FC| ≥ 1.</p>
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<p>A heatmap displaying maize chitinase gene expression under different biotic stress conditions. (<b>A</b>) Gene expression profiles of maize chitinase genes when subjected to black tassel disease stress. The experimental setup included a control treatment (uninfected plants) and treatments with the strains SG 200 and UMAG_02297, which are live nutrient fungi responsible for causing black tassel disease in maize. Additionally, the knockout mutant strain, KO_UMAG_02297, was included. Data were taken 3 days after infection (3 dpi) on six different types of maize: B73, CML 322, EGB, Ky 21, Oh 43, and Tx 303. (<b>B</b>) The expression patterns of maize chitinase genes under stem rot stress. Two maize varieties, A08 (resistant) and K09 (susceptible), were infested with <span class="html-italic">Fusarium graminearum</span>. The control treatment (CT) consisted of uninfected plants. Plants with pests were examined at different intervals including 6 h after infestation (hpi), 12 hpi, 24 hpi, 48 hpi, and 72 hpi. (<b>C</b>) Furthermore, the expression patterns of maize chitinase genes in response to maize gray spot disease stress were analyzed in two different varieties, Y (‘Yayu 889’) and Z (‘Zhenghong 532’); Y and Z are resistant and susceptible to gray spot disease, respectively. Gray spot disease in these plants was evaluated at 81, 89, 91, and 93 days post-infection (dpi). (<b>D</b>) The maize chitinase gene family’s response to sugarbeet nightshade moth and Asian corn borer infestation was analyzed. The control group (CT) consists of healthy plants, with 1 hpi, 4 hpi, 6 hpi, and 24 hpi indicating 1, 4, 6, and 24 h post-infestation, respectively. (<b>E</b>) The temporal expression of maize chitinase genes following Asian corn borer attack, with time points at 0 hpi, 4 hpi, 12 hpi, and 24 hpi corresponding to 0, 4, 12, and 24 h post-infestation intervals. (<b>F</b>) Maize chitinase gene family expression patterns during aphid stress. CT-0 h vs. CT-96 h correspond to 0 h vs. 96 h control treatments (uninfected plants); Aphid-2 h, Aphid-4 h, Aphid-8 h, Aphid-24 h, Aphid-48 h, and Aphid-96 h correspond to 2 h, 4 h, 8 h, 24 h, 48 h, and 96 h after aphid infestation, respectively. Note: Each chart depicts the original FPKM value in the left box and the value of log2(multiple changes) in the right box, highlighted in red (up-regulated) or green (down-regulated), |FC| ≥ 1.</p>
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<p>Heatmap depicting the different expression levels of maize chitinase genes in response to abiotic and biotic stresses. Here, gray denotes unchanged expression, red signifies upregulation, green indicates downregulation, and yellow represents a mix of both upregulation and downregulation.</p>
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14 pages, 8908 KiB  
Article
CaARP1/CaSGT1 Module Regulates Vegetative Growth and Defense Response of Pepper Plants against Phytophthora capsici
by Xia Li, Yahong Weng, Yufeng Chen, Kaisheng Liu, Yanyan Liu, Kan Zhang, Lanping Shi, Shuilin He and Zhiqin Liu
Plants 2024, 13(20), 2849; https://doi.org/10.3390/plants13202849 - 11 Oct 2024
Viewed by 378
Abstract
Pepper (Capsicum annuum L.) suffers severe quality and yield loss from oomycete diseases caused by Phytophthora capsici. CaSGT1 was previously determined to positively regulate the immune response of pepper plants against P. capsici, but by which mechanism remains elusive. In [...] Read more.
Pepper (Capsicum annuum L.) suffers severe quality and yield loss from oomycete diseases caused by Phytophthora capsici. CaSGT1 was previously determined to positively regulate the immune response of pepper plants against P. capsici, but by which mechanism remains elusive. In the present study, the potential interacting proteins of CaSGT1 were isolated from pepper using a yeast two-hybrid system, among which CaARP1 was determined to interact with CaSGT1 via bimolecular fluorescence complementation (BiFC) and microscale thermophoresis (MST) assays. CaARP1 belongs to the auxin-repressed protein family, which is well-known to function in modulating plant growth. The transcriptional and protein levels of CaARP1 were both significantly induced by infection with P. capsici. Silencing of CaARP1 promotes the vegetative growth of pepper plants and attenuates its disease resistance to P. capsici, as well as compromising the hypersensitive response-like cell death in pepper leaves induced by PcINF1, a well-characterized typical PAMP from P. capsici. Chitin-induced transient expression of CaARP1 in pepper leaves enhanced its disease resistance to P. capsici, which is amplified by CaSGT1 co-expression as a positive regulator. Taken together, our result revealed that CaARP1 plays a dual role in the pepper, negatively regulating the vegetative growth and positively regulating plant immunity against P. capsici in a manner associated with CaSGT1. Full article
(This article belongs to the Section Plant Protection and Biotic Interactions)
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Figure 1
<p>Confirmation of CaARP1/CaSGT1 interaction by bimolecular fluorescent complementary and microscale thermophoresis assays. (<b>A</b>) BiFC assay revealed the interaction between CaARP1 and CaSGT1. <span class="html-italic">Agrobacterium</span> cell carrying CaARP1-YFP<sup>N</sup> and CaSGT1-YFP<sup>C</sup> was co-infiltrated into the leaves of <span class="html-italic">N. benthamiana</span> using a needleless syringe. At 36–48 h post infiltration, the infiltrated leaves were sampled for the detection of the YFP signal using a confocal microscope. Bar = 25 μm. (<b>B</b>) Microscale thermophoresis experiments demonstrated that CaARP1 directly interacts with CaSGT1. Purified GST-CaSGT1 protein expressed from <span class="html-italic">E. coli</span> and purified CaARP1-GFP expressed from <span class="html-italic">N. benthamiana</span> were used in this experiment. The experiments were repeated thrice with similar results.</p>
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<p>Alignments of amino acid sequence for CaARP1 protein and its orthologs from other plant species. Alignments of amino acid sequence of CaARP1 and its orthologs from <span class="html-italic">Nicotiana tabacum</span> (NtARP1), <span class="html-italic">Solanum lycopersicum</span> (SlARP1), <span class="html-italic">Solanum stenotomum</span> (StARP1), <span class="html-italic">Arabidopsis thaliana</span> (AtARP1), <span class="html-italic">Glycine max</span> (ZmARP2), <span class="html-italic">Oryza sativa</span> (OsDAAP1) and others.</p>
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<p>Expression profiles of pepper <span class="html-italic">CaARP1</span> and <span class="html-italic">CaSGT1</span> in response to <span class="html-italic">P. capsici</span> inoculation. (<b>A</b>,<b>B</b>) Quantitative RT-PCR assay showed that the transcript accumulations of <span class="html-italic">CaARP1</span> (<b>A</b>) and <span class="html-italic">CaSGT1</span> (<b>B</b>) were both up-regulated in pepper leaves challenged with <span class="html-italic">P. capsici</span>. Leaves of pepper plants were inoculated with the spores of <span class="html-italic">P. capsici</span>, and the inoculated leaves were harvested for total RNA extraction at different time-points. The transcript accumulations of <span class="html-italic">CaARP1</span> in pepper leaves without the inoculation of <span class="html-italic">P. capsici</span> were calibrated to a standardized expression level, designated as “1” for relative comparison. Green hollow dots represent four biological replicates from one experiment. The pepper <span class="html-italic">CaACTIN</span> was used to normalize the tested genes. Asterisks indicate significant difference as determined by one-way ANOVA with Tukey’s test (* <span class="html-italic">p</span> &lt; 0.05, ** <span class="html-italic">p</span> &lt; 0.01, *** <span class="html-italic">p</span> &lt; 0.001). The experiments were replicated twice, yielding consistent and comparable outcomes.</p>
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<p>Subcellular localization of CaARP1 in leaves of <span class="html-italic">N. benthamiana</span> upon <span class="html-italic">P. capsici</span> inoculation. The leaves of <span class="html-italic">N. benthamiana</span> were transiently transformed with CaARP1-GFP or empty vector, and the <span class="html-italic">N. benthamiana</span> plants were maintained in growth chamber for 24 h, followed by the inoculation of <span class="html-italic">P. capsici</span> in the leaves. At 24 h post inoculation, the leaves were harvested for the detection of YFP signal. Empty: Empty vector (35S:<span class="html-italic">GFP</span>). Bars = 50 μm.</p>
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<p>The effect of <span class="html-italic">CaARP1</span> silencing on the vegetative growth and disease resistance of pepper plants. (<b>A</b>) The silencing efficiency of <span class="html-italic">CaARP1</span> was determined by quantitative RT-PCR. The leaves of <span class="html-italic">CaARP1</span>-silenced and unsilenced pepper leaves at 14 days post VIGS assays were sampled for total RNA extraction. The transcript level of <span class="html-italic">CaARP1</span> in unsilenced pepper plants was set to a relative expression of “1”, and <span class="html-italic">CaACTIN</span> was used to normalize <span class="html-italic">CaARP1</span> expression. (<b>B</b>) Knock-down of <span class="html-italic">CaARP1</span> expression inhibited the vegetative growth of pepper plants. At 21 days post VIGS assays, the <span class="html-italic">CaARP1</span>-silenced and unsilenced pepper plants were subjected to observation for vegetative growth. (<b>C</b>) The height of pepper plants was suppressed by <span class="html-italic">CaARP1</span> silencing. (<b>D</b>,<b>E</b>) The disease resistance of pepper leaves against <span class="html-italic">P. capsici</span> was impaired by the down-regulation of <span class="html-italic">CaARP1</span>. The phenotype was captured at 4 dpi with <span class="html-italic">P. capsici</span> spores. (<b>F</b>) The inhibitive effect of <span class="html-italic">CaARP1</span> silencing on the disease resistance of pepper in response to <span class="html-italic">P. capsici</span> was also confirmed by soil-drenching inoculation. The phenotype was captured at 7 dpi. (<b>G</b>) The HR-like cell death in <span class="html-italic">CaARP1</span>-silenced and unsilenced pepper leaves transformed with <span class="html-italic">PcINF1</span>, an elicitin from <span class="html-italic">P. capsici</span>, confirmed by phenotype (<b>left</b>), trypan blue (<b>middle</b>) and DAB (<b>right</b>) staining. Asterisks indicate significant difference as determined by one-way ANOVA with Tukey’s test (* <span class="html-italic">p</span> &lt; 0.05, ** <span class="html-italic">p</span> &lt; 0.01). The experiments were replicated three times, yielding consistent and comparable outcomes.</p>
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<p>Transient expression of <span class="html-italic">CaARP1</span> enhanced the disease resistance of pepper leaves, which was amplified by <span class="html-italic">CaSGT1</span> expression. (<b>A</b>) <span class="html-italic">CaARP1</span> expression enhanced the disease resistance of pepper leaves challenged with <span class="html-italic">P. capsici</span>, and the resistance was enhanced by the transient expression of <span class="html-italic">CaSGT1</span>. (<b>B</b>) The necrotic lesion:leaf size ratio of pepper plants inoculated with <span class="html-italic">P. capsici</span>. (<b>A</b>,<b>B</b>) The leaves of pepper plants were transiently transformed with <span class="html-italic">CaARP1</span>, <span class="html-italic">CaSGT1</span>, or empty vector, and the leaves were sprayed with chitin to induce the expression of <span class="html-italic">CaARP1</span> and <span class="html-italic">CaSGT1</span>. At 24 h post spray, the leaves were inoculated with spores of <span class="html-italic">P. capsici</span>. Black hollow dots represent three biological replicates from one experiment. Different letters indicate significant difference as determined by one-way ANOVA with Tukey’s test (<span class="html-italic">p</span> &lt; 0.05). The experiments were replicated three times, yielding consistent and comparable outcomes.</p>
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12 pages, 1494 KiB  
Article
Chitin Extracted from Black Soldier Fly Larvae at Different Growth Stages
by Andrea Marangon, Geo Paul, Riccardo Zaghi, Leonardo Marchese and Giorgio Gatti
Polymers 2024, 16(20), 2861; https://doi.org/10.3390/polym16202861 - 10 Oct 2024
Viewed by 401
Abstract
The black soldier fly (BSF) Hermetia Illucens can grow rapidly and on a wide variety of organic materials, and it is extensively used as a means of disposing of household organic waste. Different phases of the life cycle of BSF larvae (BSFL) are [...] Read more.
The black soldier fly (BSF) Hermetia Illucens can grow rapidly and on a wide variety of organic materials, and it is extensively used as a means of disposing of household organic waste. Different phases of the life cycle of BSF larvae (BSFL) are used in this work to extract chitin after the removal of lipids, mineral salts, and proteins. Multiple techniques, such as X-ray diffractometry, infrared spectroscopy, solid-state Nuclear Magnetic Resonance (13C ss-NMR) and thermogravimetric analysis, are used to investigate the chemical and physical characteristics of the extracted samples of chitin, which shows a high degree of acetylation (from 78% to 94%). The extracted chitin shows an increase of the thermal stability of 20 °C in the initial stage of life and 35 °C at the end of the life cycle if compared with a commercial standard. Moreover, the extracted chitin shows an increase in the crystallinity degree during the BSFL growth time (from 72% to 78%). Full article
(This article belongs to the Special Issue Natural Polysaccharides: Chitosan, Chitin, Pectin and Gums)
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Figure 1
<p>Schematic representation of both chitin, where R = COCH<sub>3</sub> (acetyl group) is greater than 60%, and chitosan, where R = H is greater than 60%.</p>
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<p>XRD of BSFL samples at the start and at the end of their life cycle; BSFL-IL (a) and BSFL-EL (b), respectively, compared with a commercial chitin (c).</p>
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<p>ATR-FTIR spectra of chitin samples extracted from BSFL at the start and at the end of their life cycle: BSFL-IL (a) and BSFL-EL (b), respectively. The spectrum of a commercial chitin (c) is reported for comparison.</p>
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<p><sup>13</sup>C CPMAS NMR spectra of the extracted chitin at the start and at the end of the BSFL life cycle for BSFL-IL (a) and BSFL-EL (b), respectively, compared with a commercial chitin (c). * <sup>13</sup>C peaks due to the methylene units of lipids.</p>
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<p>Thermograms (TG) and respective first derivative curves (DTG) of the extracted chitin samples BSFL-IL (a, a′) and BSFL-EL (b, b′), compared with a commercial chitin (c, c′).</p>
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16 pages, 5818 KiB  
Article
Biomineralization Process Inspired In Situ Growth of Calcium Carbonate Nanocrystals in Chitosan Hydrogels
by Xinyue Zeng, Zheng Zhu, Wei Chang, Bin Wu and Wei Huang
Appl. Sci. 2024, 14(20), 9193; https://doi.org/10.3390/app14209193 - 10 Oct 2024
Viewed by 757
Abstract
Biological composites such as bone, nacre, and teeth show excellent mechanical efficiency because of the incorporation of biominerals into the organic matrix at the nanoscale, leading to hierarchical composite structures. Adding a large volume of ceramic nanoparticles into an organic molecular network uniformly [...] Read more.
Biological composites such as bone, nacre, and teeth show excellent mechanical efficiency because of the incorporation of biominerals into the organic matrix at the nanoscale, leading to hierarchical composite structures. Adding a large volume of ceramic nanoparticles into an organic molecular network uniformly has been a challenge in engineering applications. However, in natural organisms, biominerals grow inside organic fibers, such as chitin and collagen, forming perfect ceramic/polymer composites spontaneously via biomineralization processes. Inspired from these processes, the in situ growth of calcium carbonate nanoparticles inside the chitosan network to form ceramic composites was proposed in the current work. The crystal growth of CaCO3 nanoparticles in the chitosan matrix as a function of time was investigated. A weight percentage of ~35 wt% CaCO3 composite was realized, resembling the high weight percentage of mineral phase in bones. Scanning and transmission electron microscopy indicated the integration of CaCO3 nanocrystals with chitosan macromolecules. By growing CaCO3 minerals inside the chitosan matrix, the elastic modulus and tensile strength increases by ~110% and ~90%, respectively. The in situ crystal growth strategy was also demonstrated in organic frameworks prepared via 3D printing, indicating the potential of fabricating ceramic/polymer composites with complicated structures, and further applications in tissue engineering. Full article
(This article belongs to the Section Chemical and Molecular Sciences)
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<p>The design strategy, preparation process, and microstructural characteristics of chitosan/CaCO<sub>3</sub> composites. (<b>a</b>) Schematic diagram illustrating the hierarchical structure of the natural skeleton. (<b>b</b>) Schematic representation of the compositional components of the mantis shrimp’s dactyl club. (<b>c</b>) Preparation method of the chitosan/CaCO<sub>3</sub> composite hydrogel and a schematic diagram of its structure.</p>
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<p>Microstructures of chitosan hydrogel before and after mineralization. (<b>a</b>) Images of 3D printed pure chitosan hydrogel samples at their unmineralized state. (<b>b</b>) Images of chitosan/CaCO<sub>3</sub> composite hydrogel samples after 12 h of mineralization. (<b>c</b>) SEM images of the surface of pure chitosan hydrogel at its unmineralized state. (<b>d</b>) SEM image of the chitosan/CaCO<sub>3</sub> composite hydrogel surface after 12 h of mineralization. (<b>e</b>) SEM images of the cross-section of unmineralized chitosan hydrogel. (<b>f</b>) SEM image of the cross-section of the chitosan/CaCO<sub>3</sub> composite hydrogel after 12 h of mineralization. Nanoparticles are noticed.</p>
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<p>Testing and analysis of chitosan/CaCO<sub>3</sub> composite hydrogels. (<b>a</b>) XRD test curve of the chitosan/CaCO<sub>3</sub> composite hydrogel. (<b>b</b>) TGA of chitosan/CaCO<sub>3</sub> composite hydrogel. (<b>c</b>) Raman spectral profile of chitosan/CaCO<sub>3</sub> composite hydrogels. (<b>d</b>) FTIR analysis of hydrogel before and after mineralization. (<b>e</b>) WAXD diffractogram of unmineralized pure chitosan hydrogel. (<b>f</b>) WAXD diffractogram of chitosan/CaCO<sub>3</sub> composite hydrogel.</p>
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<p>SEM images of chitosan/CaCO<sub>3</sub> composite hydrogels after 5 min and 1 h mineralization. (<b>a</b>–<b>c</b>) show SEM images of the chitosan/CaCO<sub>3</sub> composite hydrogel cross-sections after 5 min of mineralization, with (<b>b</b>) representing the area near the surface and (<b>c</b>) representing the area near the center. (<b>d</b>–<b>f</b>) show SEM images of the chitosan/CaCO<sub>3</sub> composite hydrogel cross-sections after 1 h of mineralization, with (<b>e</b>) representing the area near the surface and (<b>f</b>) representing the area near the center.</p>
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<p>SEM images of chitosan/CaCO<sub>3</sub> composite hydrogel after 3 and 6 h mineralization. (<b>a</b>–<b>c</b>) depict the cross-sections after 3 h of mineralization, where (<b>b</b>) is near the surface and (<b>c</b>) is near the center. (<b>d</b>–<b>f</b>) present the cross-sections after 6 h of mineralization, with (<b>e</b>) representing the area near the surface and (<b>f</b>) representing the area near the center.</p>
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<p>TEM images of chitosan/CaCO<sub>3</sub> composite hydrogel. (<b>a</b>,<b>b</b>) TEM image of nanoparticles formed via in situ growth. (<b>c</b>–<b>f</b>) TEM images and diffraction patterns of chitosan/CaCO<sub>3</sub> composite hydrogel. Nanoparticles shows single crystal-like diffraction pattern. Amorphous regions are noticed inside and on the edge of the calcite nanoparticles. (<b>g</b>,<b>h</b>) TEM images of the nanocomposite coating in mantis shrimp dactyl clubs, showing the combination of chitin macromolecules and hydroxyapatite nanocrystals. Images are taken from [<a href="#B16-applsci-14-09193" class="html-bibr">16</a>] with permission.</p>
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<p>Schematic of in situ growth of CaCO<sub>3</sub> nanocrystals inside chitosan hydrogels via the double diffusion method. Calcium ions are diffused into the 3D-printed chitosan hydrogel first, until an equilibrium is reached. Samples are then immersed in NaHCO<sub>3</sub> solution. As the CO<sub>3</sub><sup>2−</sup> is introduced, calcite crystals start growing at the surface first. Microcrystals are formed on the surface, while crystal sizes are limited to ~100 nm inside the hydrogel due to the confinement of the chitosan molecular network.</p>
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<p>Stress–strain curve of chitosan/CaCO<sub>3</sub> composite hydrogel at different mineralization times. Compared to the unmineralized chitosan films, mineralized films show an increase in both stiffness and tensile strength.</p>
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19 pages, 1578 KiB  
Article
Efficacy of Food Industry By-Product β-Glucan/Chitin–Chitosan on Lipid Profile of Overweight and Obese Individuals: Sustainability and Nutraceuticals
by Victoria Santisteban, Natàlia Muñoz-Garcia, Anallely López-Yerena, Montserrat Puntes, Lina Badimon and Teresa Padro
Nutrients 2024, 16(19), 3420; https://doi.org/10.3390/nu16193420 - 9 Oct 2024
Viewed by 478
Abstract
Fat-binding nutraceutical supplements have gained considerable attention as potential cholesterol-lowering strategies to address dyslipidemia in overweight and obese individuals. This study aimed to evaluate the effects of a polysaccharide-rich compound containing β-glucan/chitin–chitosan (βGluCnCs) on lipid profiles and lipoprotein function. In a prospective, two-arm [...] Read more.
Fat-binding nutraceutical supplements have gained considerable attention as potential cholesterol-lowering strategies to address dyslipidemia in overweight and obese individuals. This study aimed to evaluate the effects of a polysaccharide-rich compound containing β-glucan/chitin–chitosan (βGluCnCs) on lipid profiles and lipoprotein function. In a prospective, two-arm clinical trial, 58 overweight and obese individuals were randomized to receive either 3 g/day of βGluCnCs or a placebo (microcrystalline cellulose) for 12 weeks. Serum lipids and lipoprotein functions were assessed at baseline and at 4-week intervals throughout the study. The administration of βGluCnCs led to a significant increase in HDL cholesterol (HDLc) levels and improved HDLc/non-HDLc and HDLc/total cholesterol (TC) ratios, while reducing apolipoprotein B (ApoB) levels (p < 0.05). However, the intervention did not affect HDL particle diameter, particle number, or lipoprotein functionality. Women demonstrated greater sensitivity to changes in HDLc during βGluCnCs supplementation, whereas men exhibited a significant reduction in ApoB levels. When stratified by baseline LDL cholesterol (LDLc) levels (cut-off: 130 mg/dL), the increase in HDLc and the ApoA1/ApoB ratio was found in the low-LDL group. In contrast, the high-LDL group experienced a significant reduction in atherogenic non-LDLc and LDLc, along with an improvement in HDL’s antioxidant capacity after βGluCnCs intervention. These changes were not statistically significant in the placebo group. In conclusion, our study demonstrated that daily supplementation with βGluCnCs significantly improved lipid profiles, with effects that varied based on sex and baseline LDLc levels. Full article
(This article belongs to the Section Nutrition in Women)
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<p>Study design.</p>
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<p>Mean changes at weeks 4, 8 and 12 with respect to baseline (∆) of serum HDLc in the βGluCnCs and placebo groups. <span class="html-italic">p</span>-Value: Paired Samples <span class="html-italic">t</span> Test. <span class="html-italic">p</span>-Value *: Repeated measures analysis of variance. Statistical significance: <span class="html-italic">p</span> &lt; 0.05.</p>
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<p>Individual average changes at week 4, 8 and 12 with respect to baseline of serum HDLc, and ratios of HDLc/non-HDLc and HDLc/TC in women and men of βGluCnCs groups. Vertical spotted line represents the mean of the average changes at weeks 4, 8 and 12 with respect to baseline of βGluCnCs group. Table presents the mean of the average changes (mean ± SEM) at weeks 4, 8 and 12 with respect to baseline of men and women of βGluCnCs group. The arrows indicate the percentage of subjects with a positive increase in these variables. <span class="html-italic">p</span>-Value: one sample <span class="html-italic">t</span>-test. Statistical significance: <span class="html-italic">p</span> &lt; 0.05.</p>
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<p>Effect of the βGluCnCs in HDLc and ApoA1/ApoB levels during intervention period. Line chart graphics represent mean changes at week 4, 8 and 12 with respect to baseline (∆) of serum HDLc levels of subjects with BMI lower than 30 kg/m<sup>2</sup> or BMI equal or higher than 30 kg/m<sup>2</sup> (<b>A</b>) and subjects with low-LDLc (LDLc &lt; 130 mg/dL) and high-LDLc (LDLc ≥ 130 mg/dL) levels at baseline (<b>B</b>). Bar graphic represents ApoA1/ApoB (<b>C</b>) mean change at week 12 with respect to baseline of subjects with BMI lower than 30 kg/m<sup>2</sup> and with BMI equal or higher than 30 kg/m<sup>2</sup> and subjects with baseline low-LDLc (LDLc &lt; 130 mg/dL) and high-LDLc (LDLc ≥ 130 mg/dL) levels. <span class="html-italic">p</span>-Value: one sample <span class="html-italic">t</span>-test. Statistical significance: <span class="html-italic">p</span> &lt; 0.05. <span class="html-italic">p</span>-Value *: Repeated measures analysis of variance.</p>
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<p>Association between changes in insulin concentration and HOMA-IR (given in tertiles) with HDLc levels after 12-week intervention with βGluCnCs. T1, T2 and T3 refer to the tertile levels of (<b>A</b>) plasma insulin concentration (µU/mL) and (<b>B</b>) HOMA-IR. Bars represent the change (week 12 vs. baseline) of HDLc levels by tertiles of insulin and HOMA-IR. <span class="html-italic">p</span>-Values refer to differences vs. baseline obtained by two sample paired <span class="html-italic">t</span>-test. Statistical significance: <span class="html-italic">p</span> &lt; 0.05.</p>
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15 pages, 6834 KiB  
Article
Investigation of 9 True Weevil (Curculionidae Latreille, 1802) Species for Chitin Extraction
by Zhenying Mei, Luc Vincent, Caroline R. Szczepanski, René-Paul Godeau, Pavel Kuzhir and Guilhem Godeau
Biomimetics 2024, 9(10), 608; https://doi.org/10.3390/biomimetics9100608 - 8 Oct 2024
Viewed by 428
Abstract
Chitin, the second most abundant biopolymer after cellulose, is an important resource for biosourced materials. The global demand for chitin is rapidly increasing, however, the majority of industrial chitin is sourced from crustacean shells, which may be less accessible in regions without seafood [...] Read more.
Chitin, the second most abundant biopolymer after cellulose, is an important resource for biosourced materials. The global demand for chitin is rapidly increasing, however, the majority of industrial chitin is sourced from crustacean shells, which may be less accessible in regions without seafood waste. Therefore, it is crucial to explore alternative chitin sources, such as those derived from beetles and other arthropods. This study investigated chitin extraction from nine species of Curculionidae (true weevils), which are recognized as crop pests. The extraction process and yields were described, and the isolated chitin was characterized by SEM, IR spectroscopy, elemental analysis, XRD, and ash and water content measurements. This work highlights the potential of Curculionidae as an alternative chitin source. Full article
(This article belongs to the Special Issue Chitin and Chitosan Materials for Tomorrow)
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<p>Examples of <span class="html-italic">Curculionidae</span> specimens. (<b>A</b>) <span class="html-italic">Eupholus cuvieri</span> Guérin-Méneville, 1830, (<b>B</b>) <span class="html-italic">Eupholus magnificus</span> Kirsch, 1877, (<b>C</b>) <span class="html-italic">Lixus sturmii</span> Boheman, 1836, (<b>D</b>) <span class="html-italic">Lixus gigas</span> Fairmaire, 1904, (<b>E</b>) <span class="html-italic">Lixus albicornis</span> Fairmaire, 1904, (<b>F</b>) <span class="html-italic">Holonychus saxosus</span> Coquerel, 1859, (<b>G</b>) <span class="html-italic">Pachyrhynchus gemmatus purpureus</span> Kraatz, 1888, (<b>H</b>) <span class="html-italic">Pachyrhynchus reticulatus</span> Waterhouse, 1841 and (<b>I</b>) <span class="html-italic">Sipalinus gigas</span> Fabricius, 1775.</p>
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<p>Theoretical chemical structure of chitin.</p>
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<p>Examples of SEM images (scale bar = 30 µm) observed for treated surfaces of <span class="html-italic">E. cuvieri</span> (<b>A</b>,<b>B</b>), <span class="html-italic">E. magnificus</span> (<b>C</b>,<b>D</b>).</p>
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<p>Examples of SEM images (scale bar = 30 µm) observed for treated surfaces of <span class="html-italic">P. gemmatus purpureus</span> (<b>A</b>,<b>B</b>) and <span class="html-italic">P. reticulatus</span> (<b>C</b>,<b>D</b>).</p>
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<p>Examples of SEM images (scale bar = 30 µm) observed for treated surfaces of <span class="html-italic">L. sturmii</span> (<b>A</b>,<b>B</b>), <span class="html-italic">L. gigas</span> (<b>C</b>,<b>D</b>) and <span class="html-italic">L. albicornis</span> (<b>E</b>,<b>F</b>).</p>
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<p>Examples of SEM images (scale bar = 30 µm) observed for raw surfaces of <span class="html-italic">H. saxosus</span> (<b>A</b>,<b>B</b>) and <span class="html-italic">S. gigas</span> (<b>C</b>,<b>D</b>).</p>
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<p>Examples of FTIR spectra observed for <span class="html-italic">Curculionidae</span> specimens.</p>
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<p>Examples of thermal analysis observed for <span class="html-italic">Curculionidae</span> specimens (TG: red and DTG: blue).</p>
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<p>Examples of X-ray spectra observed for <span class="html-italic">Curculionidae</span> specimens.</p>
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30 pages, 5249 KiB  
Review
Polysaccharide-Based Bioplastics: Eco-Friendly and Sustainable Solutions for Packaging
by Ashoka Gamage, Punniamoorthy Thiviya, Anuradhi Liyanapathiranage, M. L. Dilini Wasana, Yasasvi Jayakodi, Amith Bandara, Asanga Manamperi, Rohan S. Dassanayake, Philippe Evon, Othmane Merah and Terrence Madhujith
J. Compos. Sci. 2024, 8(10), 413; https://doi.org/10.3390/jcs8100413 - 8 Oct 2024
Viewed by 1109
Abstract
Over the past few decades, synthetic petroleum-based packaging materials have increased, and the production of plastics has surpassed all other man-made materials due to their versatility. However, the excessive usage of synthetic packaging materials has led to severe environmental and health-related issues due [...] Read more.
Over the past few decades, synthetic petroleum-based packaging materials have increased, and the production of plastics has surpassed all other man-made materials due to their versatility. However, the excessive usage of synthetic packaging materials has led to severe environmental and health-related issues due to their nonbiodegradability and their accumulation in the environment. Therefore, bio-based packages are considered alternatives to substitute synthetic petroleum-based packaging material. Furthermore, the choice of packing material in the food industry is a perplexing process as it depends on various factors, such as the type of food product, its sustainability, and environmental conditions. Interestingly, due to proven mechanical, gas, and water vapor barrier properties and biological activity, polysaccharide-based bioplastics show the potential to expand the trends in food packaging, including edible films or coatings and intelligent and active food packaging. Various chemical modifications, network designs, and processing techniques have transformed polysaccharide materials into valuable final products, particularly for large-scale or high-value applications. Transitioning from petroleum-based resources to abundant bio-based polysaccharides presents an opportunity to create a sustainable circular economy. The economic viability of polysaccharide-based bioplastics is determined by several factors, including raw material costs, production technologies, market demand, and scalability. Despite their potential advantages over traditional plastics, their economic feasibility is affected by continuous technological advancements and evolving market dynamics and regulations. This review discusses the structure, properties, and recent developments in polysaccharide-based bioplastics as green and sustainable food packaging materials. Full article
(This article belongs to the Special Issue Sustainable Biocomposites, Volume II)
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<p>Biodegradable and Non-Biodegradable Plastics: Classification Based on Raw Material Origin.</p>
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<p>Classification of polysaccharides based on their origin.</p>
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<p>Structure of the amylose and amylopectin.</p>
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<p>Structure of alginic acid.</p>
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<p>Chemical structure of carrageenan: (<b>a</b>) κ, (<b>b</b>) iota, (<b>c</b>) lambda.</p>
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<p>Chemical structure of (<b>a</b>) chitin and (<b>b</b>) chitosan.</p>
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<p>Chemical structures of hyaluronic acid.</p>
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<p>Chemical structures of gellan gum.</p>
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<p>Chemical structures of Xanthan gum.</p>
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<p>Sources and possible health risks of different raw materials used for biopolymer production.</p>
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<p>Bioplastic production in terms of life cycle assessment.</p>
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14 pages, 1724 KiB  
Article
A Synbiotic Combining Chitin–Glucan and Lactobacillus acidophilus NCFM Induces a Colonic Molecular Signature Soothing Intestinal Pain and Inflammation in an Animal Model of IBS
by Lena Capirchio, Christel Rousseaux, Caroline Dubuquoy, Arthur C. Ouwehand, Véronique Maquet, Salvatore Modica, Edouard Louis, Pierre Desreumaux and Jan Tack
Int. J. Mol. Sci. 2024, 25(19), 10732; https://doi.org/10.3390/ijms251910732 - 5 Oct 2024
Viewed by 603
Abstract
Chitin–glucan (CG) is a new generation of prebiotic. Lactobacillus acidophilus NCFM® (NCFM) is a probiotic with the ability to decrease abdominal pain. We evaluate the functional and molecular gastrointestinal responses to a synbiotic administration combining CG and NCFM in a rat model [...] Read more.
Chitin–glucan (CG) is a new generation of prebiotic. Lactobacillus acidophilus NCFM® (NCFM) is a probiotic with the ability to decrease abdominal pain. We evaluate the functional and molecular gastrointestinal responses to a synbiotic administration combining CG and NCFM in a rat model of long-lasting colon hypersensitivity. The intracolonic pressure was assessed during the 9-week experiment in animals receiving CG in association or not with NCFM and compared to that in Lacticaseibacillus paracasei Lpc-37®-treated animals and control rats receiving tap water. The effects of the synbiotic were evaluated using the Wallace score, the quantification of colon myeloperoxidase (MPO) and the master genes driving analgesia and inflammation. CG 1.5 alone and NCFM 109 colony forming units (CFU) alone similarly decreased the visceral pain sensitivity. Lpc-37 had no significant effect. The best profile of pain perception inhibition was obtained with the combination of CG 1.5 g and NCFM 109 CFU, confirming a synbiotic property. This synbiotic treatment significantly reduced macroscopic colonic lesions and MPO concentrations, and induced master genes involved in analgesia (CB1, CB2, MOR, PPARα), with a downregulation of inflammatory cytokines (IL-1β, TNFα) and an induction of IL-10 and PPARγ. In conclusion, CG 1.5 g + NCFM 109 CFU significantly decreased visceral pain perception and intestinal inflammation through the regulation of master genes. Full article
(This article belongs to the Section Bioactives and Nutraceuticals)
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<p>Time-related analgesic effects of chitin–glucan, <span class="html-italic">L. acidophilus</span> NCFM and Lpc-37<sup>®</sup>. Inhibition of pain perception in % at week (W) 0-3-5-7 compared to W-2 in untreated animals receiving tap water (blue), untreated animals sensitized by TNBS (black), TNBS-sensitized rats treated with chitin–glucan at 1.5 g/d (light pink), TNBS-sensitized rats treated with <span class="html-italic">L. acidophilus</span> NCFM at 10<sup>9</sup> CFU/d (dark pink), Lpc-37<sup>®</sup> (light green), TNBS-sensitized rats treated with chitin–glucan at 1.5 g/d and <span class="html-italic">L. acidophilus</span> NCFM at 10<sup>9</sup> CFU/d (red), and TNBS-sensitized rats treated with chitin–glucan at 1.5 g/d and Lpc-37<sup>®</sup> (dark green). a: <span class="html-italic">p</span> &lt; 0.05; b: <span class="html-italic">p</span> &lt; 0.01; c: <span class="html-italic">p</span> &lt; 0.001. CG: chitin–glucan.</p>
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<p>Time- and dose-related analgesic effects of the association of chitin–glucan and <span class="html-italic">L. acidophilus</span> NCFM. (<b>A</b>) Inhibition of pain perception in % at week (W) 0-3-5-7 compared to W-2 in untreated animals receiving tap water (blue), untreated animals sensitized by TNBS (black), TNBS-sensitized rats treated with chitin–glucan at 1.5 g/d (pink), TNBS-sensitized rats treated with chitin–glucan at 1.5 g/d and <span class="html-italic">L. acidophilus</span> NCFM at 10<sup>8</sup> CFU/d (light green), TNBS-sensitized rats treated with chitin–glucan at 1.5 g/d and <span class="html-italic">L. acidophilus</span> NCFM at 10<sup>9</sup> CFU/d (medium green), and TNBS-sensitized rats treated with chitin–glucan at 1.5 g/d and <span class="html-italic">L. acidophilus</span> NCFM at 10<sup>10</sup> CFU/d (dark green). (<b>B</b>) Inhibition of pain perception in % at week (W) 0-3-5-7 compared to W-2 in untreated animals receiving tap water (blue), untreated animals sensitized by TNBS (black), TNBS-sensitized rats treated with chitin–glucan at 3 g/d (purple), TNBS-sensitized rats treated with chitin–glucan at 3 g/d and <span class="html-italic">L. acidophilus</span> NCFM at 10<sup>8</sup> CFU/d (light pink), TNBS-sensitized rats treated with chitin–glucan at 3 g/d and <span class="html-italic">L. acidophilus</span> NCFM at 10<sup>9</sup> CFU/d (medium pink), and TNBS-sensitized rats treated with chitin-glucan at 3 g/d and <span class="html-italic">L. acidophilus</span> NCFM at 10<sup>10</sup> CFU/d (red). a: <span class="html-italic">p</span> &lt; 0.05; b: <span class="html-italic">p</span> &lt; 0.01; c: <span class="html-italic">p</span> &lt; 0.001. CG: chitin–glucan.</p>
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<p>Improvement in Wallace score for macroscopic inflammatory lesions and colonic levels of MPO in animals receiving the symbiotic. (<b>A</b>) Wallace score in untreated control rats (control + vehicle, white) and in rats with colitis receiving tap water (TNBS + vehicle, black) or the symbiotic (TNBS + CG 1.5 g + NCFM 10<sup>9</sup>, hatched bars). (<b>B</b>) Myeloperoxidase (MPO) levels in the colon of untreated control rats (control + vehicle, white) and in rats with colitis receiving tap water (TNBS + vehicle, black) or the symbiotic (TNBS + CG 1.5 g + NCFM 10<sup>9</sup>, hatched bars). a: <span class="html-italic">p</span> &lt; 0.05; b: <span class="html-italic">p</span> &lt; 0.01; c: <span class="html-italic">p</span> &lt; 0.001. CG: chitin–glucan.</p>
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<p>mRNA levels of analgesic-related receptors in the colon of rats treated with the symbiotic. Levels of CB1 (<b>A</b>), CB2 (<b>B</b>), MOR (<b>C</b>) and PPARα (<b>D</b>) mRNA in the colon of untreated control rats (control + vehicle, white), rats with colitis receiving tap water (TNBS + vehicle, black) and rats with colitis treated with the symbiotic (TNBS + CG 1.5 g + NCFM 10<sup>9</sup>, hatched bars). *: <span class="html-italic">p</span> &lt; 0.05, **: <span class="html-italic">p</span> &lt; 0.01. CG: chitin–glucan.</p>
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<p>mRNA levels of inflammatory-related genes in the colon of rats treated with the symbiotic. Levels of IL-β (<b>A</b>), TNFα (<b>B</b>), IL-10 (<b>C</b>) and PPARγ (<b>D</b>) mRNA in the colon of untreated control rats (control + vehicle, white), rats with colitis receiving tap water (TNBS + vehicle, black) and rats with colitis treated with the symbiotic (TNBS + CG 1.5 g + NCFM 10<sup>9</sup>, hatched bars). *: <span class="html-italic">p</span> &lt; 0.05; **: <span class="html-italic">p</span> &lt; 0.01. CG: chitin–glucan.</p>
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<p>Long-lasting visceral hypersensitivity in rats: study design. Analgesic effects of the compounds evaluated by pain threshold at week (W) -2-0-3-5-7.</p>
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20 pages, 12359 KiB  
Article
A Thermosensitive and Degradable Chitin-Based Hydrogel as a Brucellosis Vaccine Adjuvant
by Ruibao Ju, Yanjing Lu, Zhiwen Jiang, Jinhua Chi, Shuo Wang, Wanshun Liu, Yanbo Yin and Baoqin Han
Polymers 2024, 16(19), 2815; https://doi.org/10.3390/polym16192815 - 4 Oct 2024
Viewed by 511
Abstract
Brucellosis is a zoonotic infectious disease that has long endangered the development of animal husbandry and human health. Currently, vaccination stands as the most efficacious method for preventing and managing brucellosis. Alum, as the most commonly used adjuvant for the brucellosis vaccine, has [...] Read more.
Brucellosis is a zoonotic infectious disease that has long endangered the development of animal husbandry and human health. Currently, vaccination stands as the most efficacious method for preventing and managing brucellosis. Alum, as the most commonly used adjuvant for the brucellosis vaccine, has obvious disadvantages, such as the formation of granulomas and its non-degradability. Therefore, the aims of this study were to prepare an absorbable, injectable, and biocompatible hydroxypropyl chitin (HPCT) thermosensitive hydrogel and to evaluate its immunization efficacy as an adjuvant for Brucella antigens. Specifically, etherification modification of marine natural polysaccharide chitin was carried out to obtain a hydroxypropyl chitin. Rheological studies demonstrated the reversible temperature sensitivity of HPCT hydrogel. Notably, 5 mg/mL of bovine serum albumin can be loaded in HPCT hydrogels and released continuously for more than one week. Furthermore, the L929 cytotoxicity test and in vivo degradation test in rats proved that an HPCT hydrogel had good cytocompatibility and histocompatibility and can be degraded and absorbed in vivo. In mouse functional experiments, as adjuvants for Brucella antigens, an HPCT hydrogel showed better specific antibody expression levels and cytokine (Interleukin-4, Interferon-γ) expression levels than alum. Thus, we believe that HPCT hydrogels hold much promise in the development of adjuvants. Full article
(This article belongs to the Section Biobased and Biodegradable Polymers)
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<p>The synthesis route (<b>A</b>), FT-IR spectra (<b>B</b>), and <sup>1</sup>H NMR spectra (<b>C</b>) of HPCT.</p>
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<p>(<b>A</b>) Phase transition of HPCT hydrogels with different concentrations under vial tilting method. (<b>B</b>) Plasticity and injectability of HPCT hydrogel (0.8%). (<b>C</b>) Structural morphology of HPCT hydrogel under scanning electron microscope.</p>
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<p>Rheological Behavior of HPCT. (<b>A</b>) Dynamic strain scanning of HPCT to determine the linear viscoelastic zone region (LVR). (<b>B</b>–<b>E</b>) Temperature dependence of storage modulus (G′) and loss modulus (G″) of HPCT hydrogels with different concentrations. (<b>F</b>) The correlation between viscosity and shear rate of HPCT hydrogel at 37 °C. (<b>G</b>) Viscosity changes in HPCT hydrogel at different temperatures. (<b>H</b>) Correlation between dynamic frequency (ω) and modulus of HPCT hydrogels at 37 °C. (<b>I</b>) The reversible temperature phase transition ability of HPCT.</p>
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<p>Hemolysis activity and hemolysis assay images of HPCT at different concentrations with pure water as a positive control and saline as a negative control group: (<b>A</b>) Tests using whole blood. (<b>B</b>) Tests using 2% red blood cells (Numbers 1–5 in (<b>A</b>,<b>B</b>) represent 50 μ/mL, 500 μg/mL, and 1000 μg/mL of hydrogel, saline, and pure water, respectively). Data represent the mean ± SD (<span class="html-italic">n</span> = 5). Effect of HPCT solution on the morphology and proliferation of L929 cells: (<b>C</b>) MTT test measured the proliferation rate. (<b>D</b>) Morphological changes of L929 cells were examined under a light microscope (original magnification, 100×) at 24 h, 48 h, and 72 h. Data represent the mean ± SD; <span class="html-italic">n</span> = 6; * <span class="html-italic">p</span> &lt; 0.05 (significant difference compared with the control group).</p>
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<p>(<b>A</b>) The initial state of the alum and HPCT hydrogels after subcutaneous injection into rats. (<b>B</b>) H&amp;E staining images at different times (the green pentagram areas are the subcutaneous residual material, the blue arrows show the approximate boundaries of residual material and tissue, and the black lines are the boundaries of the different areas).</p>
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<p>(<b>A</b>) The degradation curve of the HPCT hydrogel in different concentrations of lysozyme. (<b>B</b>) Percentage of BSA released in stages from the HPCT hydrogels (weight of BSA released per stage/total weight of BSA). (<b>C</b>) Total release rate for BSA from the HPCT hydrogel.</p>
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<p>(<b>A</b>) Mouse immune program diagram. (<b>B</b>) Specific antibody expression levels in mouse serum at different time points. Data represent mean ± SD, <span class="html-italic">n</span> = 8, * <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 (significant difference compared to other groups).</p>
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<p>The expression levels of cytokines IL-4 (<b>A</b>) and IFN-γ (<b>B</b>) in serum at different times after immunization. Data represent mean ± SD, <span class="html-italic">n</span> = 8, * <span class="html-italic">p</span> &lt; 0.05 and ** <span class="html-italic">p</span> &lt; 0.01 (significant difference compared to other groups).</p>
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<p>(<b>A</b>) HPCT preparation and characterization. (<b>B</b>) Hydrogel containing brucellosis antigen for immunization of mice.</p>
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12 pages, 3029 KiB  
Article
The Chitinous Skeleton of Ianthella basta Marine Demosponge as a Renewable Scaffold-Based Carrier of Antiseptics
by Izabela Dziedzic, Kamil Dydek, Alona Voronkina, Valentin Kovalchuk, Teofil Jesionowski and Hermann Ehrlich
Polysaccharides 2024, 5(4), 540-551; https://doi.org/10.3390/polysaccharides5040034 - 1 Oct 2024
Viewed by 368
Abstract
The chitinous skeleton of the marine demosponge Ianthella basta exhibits a unique network-like 3D architecture, excellent capillary properties, and chemical inertness, making it highly suitable for interdisciplinary research, especially in biomedical applications. This study investigates the potential of renewable I. basta chitinous scaffolds [...] Read more.
The chitinous skeleton of the marine demosponge Ianthella basta exhibits a unique network-like 3D architecture, excellent capillary properties, and chemical inertness, making it highly suitable for interdisciplinary research, especially in biomedical applications. This study investigates the potential of renewable I. basta chitinous scaffolds for drug delivery and wound dressing. The scaffolds, characterized by a microtubular structure, were impregnated with selected commercially available antiseptics, including solutions with hydrophilic and hydrophobic properties. Evaluations against selected clinical strains of bacteria, as well as fungi, demonstrated significant zones of growth inhibition with antiseptics such as brilliant green, gentian violet, decamethoxine, and polyhexanide. Notably, the antibacterial properties of these antiseptic-treated chitin matrices persisted for over 72 h, effectively inhibiting microbial growth in fresh cultures. These findings highlight the considerable potential of I. basta chitin scaffolds as sustainable, innovative biomaterials for controlled drug release and wound dressing applications. Full article
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<p>Dried <span class="html-italic">Ianthella basta</span> demosponge specimen on a rock fragment as collected (<b>a</b>) and a digital microscope image of its surface (<b>b</b>).</p>
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<p>(<b>a</b>) Dried <span class="html-italic">I. basta</span> sponge before chitin isolation; (<b>b</b>) <span class="html-italic">I. basta</span> sponge skeleton after treatment with acetic acid and sodium hydroxide solution; (<b>c</b>) fragment of the resulting purified chitin scaffold.</p>
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<p>Schematic diagram of the procedure for investigation of antimicrobial properties.</p>
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<p>Examples of Petri dishes with growth inhibition zones for (<b>a</b>) clinical strain of <span class="html-italic">S. aureus</span>: <b>1</b>—control, <b>2</b>—chitinous matrix impregnated with decamethoxine water solution, and <b>3</b>—chitinous matrix impregnated with decamethoxine alcohol solution; (<b>b</b>) clinical strain of <span class="html-italic">E. coli</span>, <b>1</b>—chitinous matrix impregnated with gentian violet, <b>2</b>—chitinous matrix impregnated with decamethoxine alcohol solution, and <b>3</b>—chitinous matrix impregnated with brilliant green; (<b>c</b>) clinical strain of <span class="html-italic">C. albicans</span>, <b>1</b>—chitinous matrix impregnated with solution of polyhexanide, <b>2</b>—chitinous matrix impregnated with brilliant green, and <b>3</b>—chitinous matrix impregnated with gentian violet.</p>
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<p>Results of the capacity test for the release of the antiseptics that exhibited the highest activity (gentian violet, brilliant green, decamethoxine water, and alcohol solutions) over 5 days. Images at 48, 72, and 96 h were obtained after moving the impregnated matrix to the fresh culture.</p>
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18 pages, 1625 KiB  
Article
Systematic Comparison of Extract Clean-Up with Currently Used Sorbents for Dispersive Solid-Phase Extraction
by Michelle Peter and Christoph Müller
Molecules 2024, 29(19), 4656; https://doi.org/10.3390/molecules29194656 - 30 Sep 2024
Viewed by 352
Abstract
Dispersive solid-phase extraction (dSPE) is a crucial step for multiresidue analysis used to remove matrix components from extracts. This purification prevents contamination of instrumental equipment and improves method selectivity, sensitivity, and reproducibility. Therefore, a clean-up step is recommended, but an over-purified extract can [...] Read more.
Dispersive solid-phase extraction (dSPE) is a crucial step for multiresidue analysis used to remove matrix components from extracts. This purification prevents contamination of instrumental equipment and improves method selectivity, sensitivity, and reproducibility. Therefore, a clean-up step is recommended, but an over-purified extract can lead to analyte loss due to adsorption to the sorbent. This study provides a systematic comparison of the advantages and disadvantages of the well-established dSPE sorbents PSA, GCB, and C18 and the novel dSPE sorbents chitin, chitosan, multi-walled carbon nanotube (MWCNT), and Z-Sep® (zirconium-based sorbent). They were tested regarding their clean-up capacity by visual inspection, UV, and GC-MS measurements. The recovery rates of 98 analytes, including pesticides, active pharmaceutical ingredients, and emerging environmental pollutants with a broad range of physicochemical properties, were determined by GC-MS/MS. Experiments were performed with five different matrices, commonly used in food analysis (spinach, orange, avocado, salmon, and bovine liver). Overall, Z-Sep® was the best sorbent regarding clean-up capacity, reducing matrix components to the greatest extent with a median of 50% in UV and GC-MS measurements, while MWCNTs had the largest impact on analyte recovery, with 14 analytes showing recoveries below 70%. PSA showed the best performance overall. Full article
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<p>Commonly used dSPE sorbents.</p>
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<p>Color intensities from raw extracts compared to clean-up extracts after dSPE. The raw column shows the colors of the extracts before clean-up, and the other columns show the colors of the extracts after clean-up with the respective dSPE sorbents. A reduction in color intensity or change in color tone represents the removal of matrix components (especially of pigments). Colors were extracted from the pictures in <a href="#app1-molecules-29-04656" class="html-app">Supplementary Material Figure S2</a>, shaped, and rearranged for better comparability.</p>
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<p>Distribution of the extinction of cleaned-up extracts at 220 nm relative to the extinction of the raw extracts, sorted by the sorbent used for clean-up across all matrices ((<b>A</b>), median indicated by the line in the box, whiskers show maximum and minimum values) and for the individual matrices ((<b>B</b>), n = 3, error bars show the relative standard deviation).</p>
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<p>Distribution of mean peak areas in TIC chromatograms (scan <span class="html-italic">m/z</span> 100–600) of cleaned-up extracts relative to raw extracts, sorted by the sorbent used for clean-up across all matrices ((<b>A</b>), median indicated by the line in the box, whiskers show maximum and minimum values) and for the individual matrices ((<b>B</b>), n = 3, error bars show the relative standard deviation).</p>
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<p>Analysis of the recovery data. For each matrix cleaned-up with each sorbent, the mean recovery across all analytes was calculated, resulting in one value per matrix and sorbent. The box plots represent the recoveries for the five different matrices, the lines represent the median recoveries, and the whiskers show the minimum and maximum values. The table containing all individual recovery values can be found in <a href="#app1-molecules-29-04656" class="html-app">Supplementary Material Table S2</a>.</p>
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<p>Rankings of seven tested sorbents regarding their performance in clean-up capacity experiments and affecting analyte recovery.</p>
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18 pages, 2208 KiB  
Article
Optimization of Enzymatic Deproteination of Northern Shrimp (Pandalus borealis) Shell Chitin Using Commercial Proteases
by Julia Pohling, Vegneshwaran Vasudevan Ramakrishnan, Abul Hossain, Sheila Trenholm and Deepika Dave
Mar. Drugs 2024, 22(10), 445; https://doi.org/10.3390/md22100445 - 28 Sep 2024
Viewed by 540
Abstract
Shrimp shells are a key source of chitin, commonly extracted through chemical methods, which may cause minor molecular damage. Nowadays, there is great interest in achieving close to zero protein content in crude chitin in order to use it for high-end markets. Therefore, [...] Read more.
Shrimp shells are a key source of chitin, commonly extracted through chemical methods, which may cause minor molecular damage. Nowadays, there is great interest in achieving close to zero protein content in crude chitin in order to use it for high-end markets. Therefore, this study optimized the enzymatic deproteination using two commercial proteases (SEB Pro FL100 and Sea-B Zyme L200) for effective and fast removal of residual protein from Northern shrimp (Pandalus borealis) shell chitin for the first time. The protein content was determined using both the Kjeldahl method and amino acid analysis using gas chromatography–mass spectrometry (GC-MS). The performance of papain (Sea B Zyme L200) was superior to fungal protease (SEB Pro FL100) for this application, and it achieved residual protein content of 2.01%, while the calculated optimum for the latter enzyme was 6.18%. A model was developed using 24 factorial design, and it was predicted that the lowest residual protein content using fungal protease and papain could be achieved at the following conditions: a pH of 4.2 and 7, and an enzyme concentration of 4 and 1.5%, respectively. Thus, the low-protein content obtained using enzymatic deproteination could be an alternative approach to the traditional methods, indicating their potential to produce premium-quality chitin. Full article
(This article belongs to the Collection Marine Polysaccharides)
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<p>SEB Pro FL100: Pareto chart of standardized effects (2<sup>4</sup> factorial design)<span class="html-italic">. p</span>-value is the probability used to assess the statistical significance of the result.</p>
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<p>SEB Pro FL100: Prediction and optimization plot for mean of protein vs. pH, temperature, and enzyme concentration.</p>
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<p>SEB Pro FL100: Residuals vs. fitted values.</p>
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<p>SEB Pro FL100: Additional experiments, Pareto chart of the standardized effects. <span class="html-italic">p</span>-value is the probability used to assess the statistical significance of the result.</p>
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<p>SEB Pro FL100: Additional experiments, main effects plot for mean of protein vs. enzyme concentration (EC) and pH.</p>
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<p>SEB Pro FL100: Multifactorial design, residuals vs. fitted.</p>
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<p>Sea-B-Zyme L200: Pareto chart of the standardized effects. <span class="html-italic">p</span>-value is the probability used to assess the statistical significance of the result.</p>
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<p>Enzymatic deproteination with Sea B Zyme L200: Main effects plot for protein (data means).</p>
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<p>Sea-B-Zyme L200: Residuals vs. fitted values.</p>
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<p>Compositional changes throughout chitin extraction (assuming lipid concentration is 0% and * indicates unpublished data).</p>
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14 pages, 542 KiB  
Review
A Review of Chitosan as a Coagulant of Health-Related Microorganisms in Water and Wastewater
by Collin Knox Coleman, Hemali H. Oza, Emily S. Bailey and Mark D. Sobsey
Environments 2024, 11(10), 211; https://doi.org/10.3390/environments11100211 - 24 Sep 2024
Viewed by 1034
Abstract
The coagulation and flocculation properties of chitosan, an organic biopolymer derived from chitin, have been researched as an alternative to synthetic polymers and inorganic metal salt coagulants currently used in water and wastewater treatment. In an effort to encourage further research into the [...] Read more.
The coagulation and flocculation properties of chitosan, an organic biopolymer derived from chitin, have been researched as an alternative to synthetic polymers and inorganic metal salt coagulants currently used in water and wastewater treatment. In an effort to encourage further research into the practical uses of chitosan as green chemistry in water and wastewater treatment and to promote the efficacious removal of microbial contaminants in drinking and wastewater, we have summarized the current state of research pertaining to the treatment of microorganisms in water and wastewater. A search of PubMed revealed 720 possible titles and abstracts, of which 44 full-text articles were identified as matching the eligibility criteria for inclusion in this systematic review. Results are presented based on the type of water matrix treated (i.e., drinking water, wastewater, and recreational waters) and a summary table providing details on the types and forms of chitosan utilized and the treatment mechanisms and processes described in the study. We find chitosan to be an effective coagulant, flocculant, and adsorbent for removing microbes from water and wastewater; some modified forms of chitosan can inactivate microbes and achieve disinfection, such as those containing metals like silver and antimicrobial chemicals like quaternary ammonium compounds or other strong oxidants, and use with filtration or electrochemical processes can achieve extensive reductions in microbes to meet performance targets of the World Health Organization. Full article
(This article belongs to the Special Issue Advanced Technologies of Water and Wastewater Treatment (2nd Edition))
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<p>Flow diagram of the literature search process.</p>
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18 pages, 337 KiB  
Review
Valorization of the Invasive Blue Crabs (Callinectes sapidus) in the Mediterranean: Nutritional Value, Bioactive Compounds and Sustainable By-Products Utilization
by Rosaria Arena, Giuseppe Renda, Giovanna Ottaviani Aalmo, Frédéric Debeaufort, Concetta Maria Messina and Andrea Santulli
Mar. Drugs 2024, 22(9), 430; https://doi.org/10.3390/md22090430 - 23 Sep 2024
Viewed by 1693
Abstract
The blue crab (Callinectes sapidus), originally from the western Atlantic Ocean, has recently spread to the Mediterranean and is now considered one of the one hundred most invasive species in that region. This opportunistic species, known for its adaptability to different [...] Read more.
The blue crab (Callinectes sapidus), originally from the western Atlantic Ocean, has recently spread to the Mediterranean and is now considered one of the one hundred most invasive species in that region. This opportunistic species, known for its adaptability to different temperatures and salinities, negatively impacts biodiversity and human activities such as fishing and tourism in the Mediterranean. However, the blue crab is gaining interest as a potential food resource due to its high nutritional value and delicate, sweet flavor. Its meat is rich in protein (14% to 30%), omega-3 fatty acids (EPA and DHA) and other essential nutrients beneficial for human health such as vitamins, and minerals. Utilizing this species in the production of new foods could help mitigate the negative impact of its invasiveness and offer economic opportunities. One challenge with this potential resource is the generation of waste. Approximately 6–8 million tonnes of crab shells are produced worldwide each year, leading to disposal problems and concerns regarding environmental sustainability. To improve economic and environmental sustainability, there is a need to valorize these residues, which are an important source of proteins, lipids, chitin, minerals, and pigments that can be processed into high-value-added products. However, especially in areas with industrial pollution, attention should be paid to the heavy metal (Cd and As) contents of blue crab shells. Studies suggest that blue crab by-products can be used in various sectors, reducing environmental impacts, promoting a circular economy, and creating new industrial opportunities. Full article
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