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20 pages, 4655 KiB  
Article
Formulation of Ready-to-Use Broccoli Extracts Rich in Polyphenols and Glucosinolates Using Natural Deep Eutectic Solvents
by Ivona Karaula, Emma Vasung, Anja Damjanović, Manuela Panić, Mia Radović, Kristina Radošević, Martina Bagović Kolić, Marina Cvjetko Bubalo and Ivana Radojčić Redovniković
Molecules 2024, 29(23), 5794; https://doi.org/10.3390/molecules29235794 (registering DOI) - 7 Dec 2024
Abstract
Broccoli is rich in biologically active compounds, especially polyphenols and glucosinolates, known for their health benefits. Traditional extraction methods have limitations, leading to a shift towards using natural deep eutectic solvents (NADESs) to create high-quality extracts with enhanced biological activity. This study focuses [...] Read more.
Broccoli is rich in biologically active compounds, especially polyphenols and glucosinolates, known for their health benefits. Traditional extraction methods have limitations, leading to a shift towards using natural deep eutectic solvents (NADESs) to create high-quality extracts with enhanced biological activity. This study focuses on preparing broccoli extracts in NADES, enriched with polyphenols and glucosinolates, without additional purification steps. Using the COSMOtherm software, the solubility of polyphenols and glucosinolates in NADESs was predicted, and five biocompatible betaine-based NADESs were prepared with glucose (B:Glc1:1 and B:Glc5:2), sucrose (B:Suc), glycerol (B:Gly), and malic acid (B:MA) as hydrogen bond donors. The resulting extracts were assessed for total polyphenol and glucosinolate content, along with antioxidant capacity, using the ORAC assay. The results demonstrated that NADES extracts contained higher polyphenol content and exhibited enhanced antioxidant effects compared to the reference ethanol extract, with B:Glc1:1 extract showing the highest performance among all the extracts tested. On the other hand, the extract based on B:MA exhibited nearly six times higher total glucosinolate content compared to the ethanol extract. Additionally, polyphenols and glucosinolates were generally more stable in NADES extracts than in the reference solvent. Finally, the B:Glc1:1 extract, identified as optimal in terms of polyphenol and glucosinolate content and stability, exhibited mild stimulation of HaCaT cells growth and facilitated the wound-healing process. Through green chemistry parameter calculations, we demonstrated that the extraction of broccoli bioactives using B:Glc1:1 can be considered sustainable, underscoring the potential of NADESs for producing ready-to-use plant extracts. Full article
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Figure 1

Figure 1
<p>Structures of quercetin, ferulic acid, and glucoraphanin used as input parameters for COSMOtherm calculations.</p>
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<p>(<b>A</b>) Total polyphenolic content in the prepared broccoli extracts. (<b>B</b>) ORAC values of the prepared extracts. Results are expressed as the means (<span class="html-italic">n</span> = 3) ± S.D.</p>
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<p>Glucosinolate profile of NADESs and ethanol (70%, <span class="html-italic">v</span>/<span class="html-italic">v</span>) extracts of broccoli. Results are presented as mean ± S.D. (<span class="html-italic">n</span> = 3).</p>
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<p>Residual concentration of total polyphenols in broccoli extracts: polyphenol content in extracts stored at 4 °C (<b>A</b>) and 25 °C (<b>B</b>), expressed as the ratio of polyphenols concentration in the extract after incubation and the initial polyphenol concentration in the extract. Results are presented as mean ± S.D. (<span class="html-italic">n</span> = 3).</p>
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<p>Residual concentration of glucosinolates in broccoli extracts: aliphatic glucosinolates in extracts stored at 25 °C (<b>A</b>) and 4 °C (<b>B</b>), and indole glucosinolates in extracts stored at 25 °C (<b>C</b>) and 4 °C (<b>D</b>), expressed as the ratio of glucosinolate concentration in the extract after incubation to the initial glucosinolate concentration in the extract. Results are presented as mean ± S.D. (<span class="html-italic">n</span> = 3).</p>
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<p>Radar plot evaluating the extracts in terms of target properties. The radar chart is bounded by the specific lower and upper limits for each target property. Ratings, ranging from 0 to 100, reflect the performance of extracts relative to the best candidate for each target property.</p>
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<p>(<b>A</b>) Effect of B:Glc<sub>1:1</sub> and corresponding extract on HaCat cell viability determined by the MTS assay in volume ratio 0.5–5% (<span class="html-italic">v</span>/<span class="html-italic">v</span>). (<b>B</b>) Migration assessment of HaCaT cells: the percentage of wound closure, determined from changes in gap width from the initial scratch over 24 and 48 h. (<b>C</b>) Microscopic images from in vitro scratch wound-healing assays showing cell migration into the cell-free gap (outlined) over time, comparing untreated cells as control with cells treated with B:Glc<sub>1:1</sub> broccoli extract and B:Glc<sub>1:1</sub> alone.</p>
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13 pages, 1317 KiB  
Article
Bioactivation, Mutagenicity, DNA Damage, and Oxidative Stress Induced by 3,4-Dimethylaniline
by Mariam R. Habil, Raúl A. Salazar-González, Mark A. Doll and David W. Hein
Biomolecules 2024, 14(12), 1562; https://doi.org/10.3390/biom14121562 (registering DOI) - 7 Dec 2024
Abstract
3,4-Dimethylaniline (3,4-DMA) is present in cigarette smoke and widely used as an intermediate in dyes, drugs, and pesticides. Nucleotide excision repair-deficient Chinese hamster ovary (CHO) cells stably transfected with human CYP1A2 and N-acetyltransferase 1 (NAT1) alleles: NAT1*4 (reference allele) or NAT1*14B (the most [...] Read more.
3,4-Dimethylaniline (3,4-DMA) is present in cigarette smoke and widely used as an intermediate in dyes, drugs, and pesticides. Nucleotide excision repair-deficient Chinese hamster ovary (CHO) cells stably transfected with human CYP1A2 and N-acetyltransferase 1 (NAT1) alleles: NAT1*4 (reference allele) or NAT1*14B (the most common variant allele) were utilized to assess 3,4-DMA N-acetylation and hypoxanthine phosphoribosyl transferase (HPRT) mutations, double-strand DNA breaks and reactive oxygen species (ROS). CHO cells expressing NAT1*4 exhibited significantly (p < 0.001) higher 3,4-DMA N-acetylation rates than CHO cells expressing NAT1*14B both in vitro and in situ. In CHO cells expressing CYP1A2 and NAT1, 3,4-DMA caused concentration-dependent increases in reactive oxygen species (ROS), double-stranded DNA damage, and HPRT mutations. CHO cells expressing NAT1*4 and NAT1*14B exhibited concentration-dependent increases in ROS following treatment with 3,4-DMA (linear trend p < 0.001 and p < 0.0001 for NAT1*4 and NAT1*14B, respectively) that were lower than in CHO cells expressing CYP1A2 alone. DNA damage and oxidative stress induced by 3,4-DMA did not differ significantly (p >0.05) between CHO cells expressing NAT1*4 and NAT1*14B. CHO cells expressing NAT1*14B showed higher HPRT mutants (p < 0.05) than CHO cells expressing NAT1*4. These findings confirm 3,4-DMA genotoxicity consistent with potential carcinogenicity. Full article
(This article belongs to the Special Issue DNA Damage, Mutagenesis, and Repair Mechanisms)
14 pages, 4938 KiB  
Article
Attenuating Oxidative Damage with Macelignan in Glutamate-Induced HT22 Hippocampal Cells
by Mei Tong He, Kiwon Jung, Chan-Woong Park, Young-Won Chin and Ki Sung Kang
Appl. Sci. 2024, 14(23), 11408; https://doi.org/10.3390/app142311408 (registering DOI) - 7 Dec 2024
Abstract
Macelignan, from Myristica fragrans (nutmeg), is a bioactive compound with various pharmacological properties, including anti-inflammatory and neuroprotective activities. The purpose of this work was to investigate the antioxidant and anti-apoptotic effects of macelignan in glutamate-treated HT22 mouse hippocampal neurons. Macelignan was extracted and [...] Read more.
Macelignan, from Myristica fragrans (nutmeg), is a bioactive compound with various pharmacological properties, including anti-inflammatory and neuroprotective activities. The purpose of this work was to investigate the antioxidant and anti-apoptotic effects of macelignan in glutamate-treated HT22 mouse hippocampal neurons. Macelignan was extracted and identified in a methanol extract of M. fragrans seeds. The DPPH was used to assess the antioxidative activity of macelignan. Glutamate (5 mM) was used to induce neurotoxicity in the HT22 cells. Neuroprotective effects were measured using relevant biochemical and imaging assays, including cell viability, ROS production, nuclear staining, apoptotic cell death, and protein expression. Macelignan markedly and concentration-dependently enhanced DPPH radical scavenging activity. In the HT22 cell model, glutamate induced cell damage by decreasing cell viability, promoting ROS generation, and increasing apoptotic cell death according to cell morphological changes. However, macelignan treatment restored cell viability, inhibited ROS generation concentration-dependently, and reduced apoptosis. Moreover, glutamate significantly up-regulated the phosphorylation of MAPK-pathway-related proteins, which was reversed by macelignan treatment. In conclusion, macelignan shows notable neuroprotective effects on oxidative stress and apoptotic cell death in glutamate-induced cells, and this study provides useful information on its potential therapeutic implications in neurological disorders. Full article
18 pages, 4884 KiB  
Article
Evaluation of the Leaves and Seeds of Cucurbitaceae Plants as a New Source of Bioactive Compounds for Colorectal Cancer Prevention and Treatment
by Mercedes Peña, Ana Guzmán, Cristina Mesas, Jesús M. Porres, Rosario Martínez, Francisco Bermúdez, Consolación Melguizo, Laura Cabeza and Jose Prados
Nutrients 2024, 16(23), 4233; https://doi.org/10.3390/nu16234233 (registering DOI) - 7 Dec 2024
Viewed by 46
Abstract
Background/Objectives: The Cucurbitaceae family represents an important source of bioactive compounds with antioxidant, antimicrobial, anti-inflammatory and antitumor activities. This study aims to investigate the potential application of Cucurbitaceae leaves and seed extracts to prevent and treat colorectal cancer (CRC). Methods: Four extracts (ethanol [...] Read more.
Background/Objectives: The Cucurbitaceae family represents an important source of bioactive compounds with antioxidant, antimicrobial, anti-inflammatory and antitumor activities. This study aims to investigate the potential application of Cucurbitaceae leaves and seed extracts to prevent and treat colorectal cancer (CRC). Methods: Four extracts (ethanol extracts and protein extracts and hydrolysates) from the leaves and seeds of cucurbits were tested in T-84, HCT-15 and HT-29 CRC cells. The antitumor, antiangiogenic, antioxidant and chemopreventive potentials and bioactive composition of the active extracts were characterized. Results: Cold ethanolic extracts from the leaves and seeds of two interspecific Cucurbita genera (CLU01002 and COK01001) exhibited potent antiproliferative, specific and non-hepatotoxic activity against CRC cell lines, with a slight synergistic effect in combination with oxaliplatin. This antitumor activity was related to G2/M cell cycle arrest, the extrinsic apoptosis pathway, cytokinesis inhibition and autophagy. The extracts also inhibited tumor clonogenicity and angiogenesis, and modulated cancer stem cell (CSC) gene expression, as well as expressing antioxidant and chemopreventive cellular capabilities. Finally, phenolic and cucurbitane-type triterpenoid compounds (pengxianencins and cucurbitacins) were tentatively identified in the active extracts by UPLC-MS analysis and bioguided fractionation. Conclusions: Extracts from leaves the and seeds of two interspecific Cucurbita genera (CLU01002 and COK01001) may contribute to the improvement of prevention and treatment strategies for CRC patients. Full article
(This article belongs to the Section Phytochemicals and Human Health)
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Figure 1

Figure 1
<p>Antitumor potential of cEtOH from the leaves (L) and seeds (S) of CLU and COK plants alone or in combination with chemotherapeutic drugs. (<b>a</b>) Relative proliferation (RP, %) of cell lines treated with cEtOH for 72 h was determined by SRB assay. Data are the mean ± SD of three replicates from two different extractions. (<b>b</b>) HSA synergy scores of T-84 cells co-treated with cEtOH and 5-FU or OXA were plotted on heatmaps with red for synergy (&gt;10), white for additive effect (from −10 to 10) and green for antagonism (&lt;−10), using SynergyFinder Plus.</p>
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<p>Cell cycle and apoptosis analysis of T-84 cells treated with cEtOH extracts from CLU and COK plants. (<b>a</b>) Cell population distribution of T-84 cells exposed to cEtOH from the leaves (L) and seeds (S) of CLU and COK for cell cycle analysis (48 and 72 h) and (<b>b</b>) for the study of apoptosis (48 h) using flow cytometry. Results are expressed as mean ± SD of three replicates. (<b>c</b>) Representative Western blot images of T-84 cells treated with cEtOH from L and S of CLU and COK for 24 h (caspase-8 and -9 and γ-H2AX) and 72 h (PARP1). WB images of PARP1 were obtained by splicing different lanes. (<b>d</b>) Relative protein expression was calculated as the mean ± SD of three measurements from three Western blot experiments. Statistical significance compared to control cells: <span class="html-italic">p</span> value &lt; 0.05 (*); &lt;0.01 (**); &lt;0.001 (***).</p>
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<p>Cellular microtubule organization and autophagy process in T-84 cells treated with cEtOH extracts from CLU and COK plants. (<b>a</b>) Representative fluorescent microscopy images of α-tubulin (upper-green, amplified from 20× magnification) and acidic vesicles (lower-red, indicated by arrows, original 40× magnification) in cells treated with cEtOH from the leaves (L) and seeds (S) of CLU and COK for 24 h. Cell nuclei (blue) were stained with Hoechst 33342. (<b>b</b>) Lysotracker/Hoechst staining ratio relative to control cells was quantified using ImageJ software as the mean ± SD of three images from four replicates. (<b>c</b>) Determination of LC3B protein expression in T-84 cells treated with cEtOH from L and S of CLU and COK (24 h) by Western blot. The LC3B-II/LC3B-I ratio was calculated as the mean ± SD of three Western blot replicates. Statistical significance compared to control cells: <span class="html-italic">p</span> value &lt; 0.001 (***).</p>
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<p>Study of clonogenicity and cell migration of T-84 cells treated with cEtOH extracts from CLU and COK plants. (<b>a</b>) Representative images of T-84 cells colonies (12 days) stained with SRB and graph representation of the percentage of colony formation. Cells were pretreated (72 h) with cEtOH (IC<sub>25</sub> and IC<sub>75</sub>) from the leaves (L) and seeds (S) of CLU and COK. (<b>b</b>) Representative light microscopy images of wound healing assay in T-84 cells treated with cEtOH from L and S of CLU and COK (4× magnification), and graphical representation of the percentage of cell migration. Data were represented as the mean ± SD of triplicate cultures. Statistical significance compared to control cells: <span class="html-italic">p</span> value &lt; 0.05 (*); &lt;0.001 (***).</p>
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<p>Study of the antiangiogenic potential of cEtOH extracts from CLU and COK plants. (<b>a</b>) Stereomicroscope images of the CAM (treatment, external and intermediate areas respect to the ring) treated with saline solution (40 μL/egg; negative control), aflibercept (4 mg/mL; positive control) and cEtOH from the leaves (L) and seeds (S) of CLU and COK (250 μg/mL) for 72 h (3× magnification). (<b>b</b>) Vascular density and vascular length density areas presented as the mean ± SD (n = 4). (<b>c</b>) Western blot of T-84 cells’ VEGFA expression after treatment (24 h) with cEtOH from L and S of CLU and COK. WB images of VEGFA were obtained by splicing different lanes. The VEGFA/β-actin ratio was presented as the mean ± SD of three measures from three Western blot experiments. Statistical significance compared to negative control: <span class="html-italic">p</span> value &lt; 0.05 (*); &lt;0.01 (**); &lt;0.001 (***).</p>
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<p>In vitro antioxidant and chemopreventive activity of cEtOH extracts from CLU and COK plants. (<b>a</b>) Percentage of relative viability of cells pretreated with cEtOH extracts (non-toxic doses) and exposed to hydrogen peroxide (H<sub>2</sub>O<sub>2</sub>). Relative viability (RV, %) was calculated by MTT assay and results are expressed as the mean ± SD of 8 replicates. The statistically significant differences were calculated compared to treated cells treated with H<sub>2</sub>O<sub>2</sub>: <span class="html-italic">p</span> value &lt; 0.01 (**); &lt;0.001 (***). (<b>b</b>) Glutathione S-Transferase (GST) and NAD(P)H quinone oxidoreductase (QR) activity in cytosolic fractions obtained from HT-29 cells treated with cEtOH. Sulforaphane (SFN) was used as positive control. Statistical significance compared to control cells: <span class="html-italic">p</span> value &lt; 0.05 (*); &lt;0.001 (***).</p>
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<p>UPLC-MS chromatographic profiles of cEtOH obtained from leaves and seeds of CLU and COK plants. The highest peaks were highlighted and tentatively identified in mass spectrometry databases such as the Dictionary of Natural Products and the Chemspider database.</p>
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28 pages, 1315 KiB  
Article
In Vitro Evaluation, Chemical Profiling, and In Silico ADMET Prediction of the Pharmacological Activities of Artemisia absinthium Root Extract
by Asma N. Alsaleh, Ibrahim M. Aziz, Reem M. Aljowaie, Rawan M. Alshalan, Noorah A. Alkubaisi and Mourad A. M. Aboul-Soud
Pharmaceuticals 2024, 17(12), 1646; https://doi.org/10.3390/ph17121646 (registering DOI) - 7 Dec 2024
Viewed by 37
Abstract
Artemisia absinthium L., is a plant with established pharmacological properties, but the A. absinthium root extract (AARE) remains unexplored. The aim of this study was to examine the chemical composition of AARE and assess its biological activity, which included antidiabetic, antibacterial, anticancer, and [...] Read more.
Artemisia absinthium L., is a plant with established pharmacological properties, but the A. absinthium root extract (AARE) remains unexplored. The aim of this study was to examine the chemical composition of AARE and assess its biological activity, which included antidiabetic, antibacterial, anticancer, and antioxidant properties. GC-MS was used to analyze the chemical components. The antioxidant activity of the total phenolic and flavonoid content was evaluated. Antibacterial activity and cytotoxic effects were identified. Enzyme inhibition experiments were performed to determine its antidiabetic potential. Molecular docking was utilized to evaluate the potential antioxidant, antibacterial, and anticancer activities of the compounds from AARE using Maestro 11.5 from the Schrödinger suite. AARE exhibited moderate antioxidant activity in DPPH (IC50: 172.41 ± 3.15 μg/mL) and ABTS (IC50: 378.94 ± 2.18 μg/mL) assays. Cytotoxicity tests on MCF-7 and HepG2 cancer cells demonstrated significant anticancer effects, with IC50 values of 150.12 ± 0.74 μg/mL and 137.11 ± 1.33 μg/mL, respectively. Apoptotic studies indicated an upregulation of pro-apoptotic genes (caspase-3, 8, 9, Bax) and a downregulation of anti-apoptotic markers (Bcl-2 and Bcl-Xl). AARE also inhibited α-amylase and α-glucosidase, suggesting potential antidiabetic effects, with IC50 values of 224.12 ± 1.17 μg/mL and 243.35 ± 1.51 μg/mL. Antibacterial assays revealed strong activity against Gram-positive bacteria. Molecular docking and pharmacokinetic analysis identified promising inhibitory effects of key AARE compounds on NADPH oxidase, E. coli Gyrase B, and Topoisomerase IIα, with favorable drug-like properties. These findings suggest AARE’s potential in treating cancer, diabetes, and bacterial infections, warranting further in vivo and clinical studies. Full article
21 pages, 6961 KiB  
Article
Composition of Human-Associated Gut Microbiota Determines 3-DF and 3-HF Anti-Colitic Activity in IL-10 -/- Mice
by Jose Haro-Reyes, Jayaprakash Kanijam Raghupathi and Lavanya Reddivari
Nutrients 2024, 16(23), 4232; https://doi.org/10.3390/nu16234232 (registering DOI) - 7 Dec 2024
Viewed by 78
Abstract
Background: Gut bacterial dysbiosis along with intestinal mucosal disruption plays a critical role in inflammatory disorders like ulcerative colitis. Flavonoids and other food bioactives have been studied in mice models as alternative treatments with minimal side effects. However, most of the research has [...] Read more.
Background: Gut bacterial dysbiosis along with intestinal mucosal disruption plays a critical role in inflammatory disorders like ulcerative colitis. Flavonoids and other food bioactives have been studied in mice models as alternative treatments with minimal side effects. However, most of the research has been carried out with mice-native microbiota, which limits the comprehension of the interaction between flavonoids and human-associated bacteria. Hence, the objective of our study was to determine the effect of healthy human-associated microbiota on the anti-colitic activity of diets rich in anthocyanins (3-HF) and phlobaphenes (3-DF). Methods: In this regard, the interleukin (IL)-10 -/- mice model was utilized. Mice were divided into three groups for inoculation with human gut bacteria from three different healthy donors and assigned to four diets. A purified diet (Diet P) and three diets containing 25% near-isogenic lines (NILs) of corn were evaluated. Diets were substituted with NILs expressing only 3-DFs (diet B), only 3-HFs (diet C), and both 3-DF and 3-HF (diet D). Results: In an overall analysis, flavonoid-rich diets did not affect inflammatory markers, microbiota diversity, or gut metabolites, but diets containing anthocyanins improved barrier function parameters. However, when data was segmented by the recipient’s microbiota from different human donors, the diet effects became significant. Furthermore, 3-HFs showed more beneficial effects than 3-DFs across the recipient’s microbiota. Conclusions: Our study suggests that the anti-colitic activity of 3-DF and 3-HF and their gut metabolites depends on the donor’s microbial composition. Full article
(This article belongs to the Special Issue Anthocyanins and Human Health—2nd Edition)
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Graphical abstract

Graphical abstract
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<p>Experimental design. Fecal microbial transplantation (FMT), near-isogenic lines (NILs), ulcerative colitis (UC). Human fecal donors: H-1, H-2, and H-3.</p>
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<p>Overall diet effects on anatomic and symptomatic parameters. Liver (<b>A</b>), spleen (<b>B</b>), kidneys (<b>C</b>), colon (<b>D</b>), cecum (<b>E</b>), and colon length (<b>F</b>) are expressed as ratios of body weight, together with the percentage of weight loss (<b>G</b>) and disease activity index (<b>H</b>). Diets: purified (P), and 25% inclusion of maize NIL with 3-DFs (B), 3-HFs (C), or both 3-DF + 3HF (D). Replicates (<span class="html-italic">n</span> = 9–12) are shown for every diet. Statistical differences are represented by (*) <span class="html-italic">p</span> ≤ 0.05, and (#) <span class="html-italic">p</span> ≤ 0.10.</p>
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<p>Diet effects on anatomic and symptomatic parameters by microbiota of donor’s recipients. Liver (<b>A</b>), spleen (<b>B</b>), kidneys (<b>C</b>), colon (<b>D</b>), cecum (<b>E</b>), and colon length (<b>F</b>) are expressed as ratios of body weight, together with the percentage of weight loss <b>G</b>) and disease activity index (<b>H</b>). Diets: purified (P), and 25% inclusion of maize NIL with 3-DFs (B), 3-HFs (C), or both 3-DF + 3HF (D). Human fecal donors: H-1, H-2, and H-3. Replicates (<span class="html-italic">n</span> = 3–5) are shown for every diet per donor. Statistical differences are represented by (*) <span class="html-italic">p</span> ≤ 0.05, and (#) <span class="html-italic">p</span> ≤ 0.10.</p>
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<p>Overall diet effects on PRRs and inflammatory markers. mRNA expression of TLR-4 (<b>A</b>), TLR-5 (<b>B</b>), NF-κB (<b>C</b>), TNF-α (<b>D</b>), IL-1β (<b>E</b>), and IL-6 (<b>F</b>). Diets: purified (P), and 25% inclusion of maize NIL with 3-DFs (B), 3-HFs (C), or both 3-DF + 3HF (D). Replicates (<span class="html-italic">n</span> = 9–12) are shown for every diet.</p>
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<p>Diet effects on PPRs and inflammatory markers by microbiota of donor’s recipients. mRNA expression of TLR-4 (<b>A</b>), TLR-5 (<b>B</b>), NF-κB (<b>C</b>), TNF-α (<b>D</b>), IL-1β (<b>E</b>), and IL-6 (<b>F</b>). Diets: purified (P), and 25% inclusion of maize NIL with 3-DFs (B), 3-HFs (C), or both 3-DF + 3HF (D). Human fecal donors: H-1, H-2, and H-3. Replicates (<span class="html-italic">n</span> = 3–5) are shown for every diet per donor. Statistical differences are represented by (*) <span class="html-italic">p</span> ≤ 0.05, and (#) <span class="html-italic">p</span> ≤ 0.10.</p>
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<p>The effect of diet on gut permeability, mucus protection, and tight junction proteins. FITC concentration in serum (<b>A</b>), mRNA expression of Muc2 (<b>B</b>), occludin (<b>C</b>), and TJP-1 (<b>D</b>). Diets: purified (P), and 25% inclusion of maize NIL with 3-DFs (B), 3-HFs (C), or both 3-DF + 3HF (D). Replicates (<span class="html-italic">n</span> = 9–12) are shown for every diet. Statistical differences are represented by (*) <span class="html-italic">p</span> ≤ 0.05, and (#) <span class="html-italic">p</span> ≤ 0.10.</p>
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<p>On the effect of diet on gut permeability, mucus protection, and tight junction proteins by the microbiota of each donor’s recipients. FITC serum concentration (<b>A</b>), mRNA expression of Muc2 (<b>B</b>), occludin (<b>C</b>), and TJP-1 (<b>D</b>). Diets: purified (P), and 25% inclusion of maize NIL with 3-DFs (B), 3-HFs (C), or both 3-DF + 3HF (D). Human fecal donors: H-1, H-2, and H-3. Replicates (<span class="html-italic">n</span> = 3–5) are shown for every diet per donor. Statistical differences are represented by (*) <span class="html-italic">p</span> ≤ 0.05, and (#) <span class="html-italic">p</span> ≤ 0.10.</p>
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<p>Overall diet effects on α-diversity indexes. Shannon index (<b>A</b>), Pielou evenness (<b>B</b>), Faith phylogenetic diversity (<b>C</b>), and observed features (<b>D</b>). Diets: purified (P), and 25% inclusion of maize NIL with 3-DFs (B), 3-HFs (C), or both 3-DF + 3HF (D). Replicates (<span class="html-italic">n</span> = 9–12) are shown for every diet.</p>
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<p>Principal coordinate analysis of β-diversity. Jaccard index (<b>A</b>) and Bray–Curtis dissimilarity (<b>B</b>), unweighted UniFrac (<b>C</b>), and weighted UniFrac (<b>D</b>) distances. Diets: purified (P), and 25% inclusion of maize NIL with 3-DFs (B), 3-HFs (C), or both 3-DF + 3HF (D). Human fecal donors: H-1, H-2, and H-3. Replicates (<span class="html-italic">n</span> = 9–12) are shown for every diet.</p>
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<p>Overall taxonomic diversity plot showing the relative abundance of gut microbiota at the family level per dietary treatment. Taxa higher than 1% relative abundance is plotted. Diets: purified (P), and 25% inclusion of maize NIL with 3-DFs (B), 3-HFs (C), or both 3-DF + 3HF (D).</p>
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<p>Taxonomic diversity plot showing diet effects in the relative abundance of gut microbiota Families by donor of microbiota. Taxa higher than 1% relative abundance is plotted. H-1, H-2, and H-3 refer to the human fecal donors. Diets: purified (P), and 25% inclusion of maize NIL with 3-DFs (B), 3-HFs (C), or both 3-DF + 3HF (D).</p>
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<p>Overall diet effect on gut metabolites: short-chain fatty acids and primary bile acids. SCFAs: acetate (<b>A</b>), propionate (<b>B</b>), butyrate (<b>C</b>), and bile acids: glycocholic acid, GCA (<b>D</b>), taurochenodeoxycholic acid, TCDCA (<b>E</b>), tauroursodeoxycholic acid, TUDCA (<b>F</b>), cholic acid, CA (<b>G</b>), chenodeoxycholic acid, CDCA (<b>H</b>), α-murocholic acid, α-MCA (<b>I</b>) and β-murocholic acid, β-MCA (<b>J</b>). Diets: purified (P), and 25% inclusion of maize NIL with 3-DFs (B), 3-HFs (C), or both 3-DF + 3HF (D). Replicates (<span class="html-italic">n</span> = 9–12) are shown for every diet. Statistical differences are represented by (*) <span class="html-italic">p</span> ≤ 0.05.</p>
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<p>Dietary effect on gut metabolites: short-chain fatty acids and primary bile acids by microbiota of donor’s recipients. SCFAs: acetate (<b>A</b>), propionate (<b>B</b>), butyrate (<b>C</b>), and bile acids: glycocholic acid, GCA (<b>D</b>), taurochenodeoxycholic acid, TCDCA (<b>E</b>), tauroursodeoxycholic, TUDCA (<b>F</b>), cholic acid, CA (<b>G</b>), chenodeoxycholic acid, CDCA (<b>H</b>), α-murocholic acid, α-MCA (<b>I</b>) and β-murocholic acid, β-MCA (<b>J</b>). Diets: purified (P), and 25% inclusion of maize NIL with 3-DFs (B), 3-HFs (C), or both 3-DF + 3HF (D). Human fecal donors: H-1, H-2, and H-3. Replicates (<span class="html-italic">n</span> = 3–5) are shown for every diet per donor. Statistical differences are represented by (*) <span class="html-italic">p</span> ≤ 0.05, and (#) <span class="html-italic">p</span> ≤ 0.10.</p>
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<p>Correlation heatmap between biomarkers and gut metabolites. Darker colors represent higher positive correlations, while lighter represent highly negative correlations. Only significant <span class="html-italic">p</span> &lt; 0.05 correlations are shown. Pattern recognition receptors: toll-like receptor-4, TLR-4, toll-like receptor-5, TLR-5. Inflammatory markers: interleukin-1β, IL-1β, interleukin 6, IL-6, tumor necrotic factor- α, TNF-α. Tight junction proteins: occludin and tight junction protein 1, TJP-1. Mucin 2: Muc2. Disease activity index: DAI. Bile acids: glycocholic acid, GCA, taurochenodeoxycholic acid, TCDCA, tauroursodeoxycholic, TUDCA, cholic acid, CA, chenodeoxycholic acid, CDCA, α-murocholic acid, α-MCA, and β-murocholic acid, β-MCA.</p>
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22 pages, 3147 KiB  
Article
Deciphering the Phytochemical Potential of Hemp Hairy Roots: A Promising Source of Cannabisins and Triterpenes as Bioactive Compounds
by Naomi Kaminsky, Jane Hubert, Cédric Guerin, Malak Mazlani, Alexis Kotland, Victor Pozzobon, Blandine Marant, Héloïse Mailhac and Stéphane Poigny
Molecules 2024, 29(23), 5792; https://doi.org/10.3390/molecules29235792 (registering DOI) - 7 Dec 2024
Viewed by 87
Abstract
Cannabis sativa L., specifically hemp, is a traditional herbaceous plant with industrial and medicinal uses. While much research has focused on cannabinoids and terpenes, the potential of hemp roots is less explored due to bioproduction challenges. Still, this material is rich in bioactive [...] Read more.
Cannabis sativa L., specifically hemp, is a traditional herbaceous plant with industrial and medicinal uses. While much research has focused on cannabinoids and terpenes, the potential of hemp roots is less explored due to bioproduction challenges. Still, this material is rich in bioactive compounds and demonstrates promising anti-inflammatory, antimicrobial, and antioxidant properties. Biotechnological methods, such as hairy root cultures, enable the efficient production of specialized metabolites while avoiding the issues of outdoors cultures. Despite these benefits, the chemical diversity understanding of hemp hairy roots remains limited. In this study, we conducted an extensive NMR and LC/MS chemical profiling of hemp hairy roots to determine their chemical composition, revealing the presence of cannabisins for the first time. We then investigated the accumulation of cannabisins and triterpenes in both hemp hairy roots and hemp aeroponic roots. Our findings reveal that hairy roots produce 12 times more cannabisins and 6 times more triterpenes than aeroponic roots, respectively, in addition to yielding 3 times more biomass in bioreactors. Preliminary bioassays also suggest antioxidant and antifungal properties. This research underscores the potential of hemp hairy roots as a valuable source of specialized metabolites and calls for further exploration into their bioactive compounds and applications. Full article
(This article belongs to the Special Issue Recent Advances in Cannabis and Hemp Research)
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<p>HHR and HAR development from 0 to 42 days of culture and harvested biomass. In vitro HHRs were grown in Erlenmeyer for 21 days, transferred to a 1 L-bioreactor (<span class="html-italic">n</span> = 3), and further cultivated until the harvest at 42 days. HARs were grown from 10-day plantlets (<span class="html-italic">n</span> = 3) and transferred to a semi-controlled indoor aeroponic system until the harvest on day 42.</p>
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<p>Chemical structures of the prevailing molecules unambiguously identified by NMR in HHR-UP and HHR-LOW.</p>
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<p>LC/MS BPI chromatogram (ESI+ and ESI−) of the extracts HHR-UP and HHR-LOW.</p>
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<p>Chemical structures of cannabisins.</p>
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<p>Chromatograms of friedelin and epifriedelanol quantification in HHR DMSO extract by GC-FID and their chemical structures.</p>
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<p>Quantification of triterpenes and cannabisins in HHRs and HARs. (<b>a</b>) Quantification of triterpenes from DMSO extracts of HHRs and HARs using GC-FID. Significantly higher amounts of triterpenes have been quantified in HHRs compared to HARs. Data are shown in mg/g. (<b>b</b>) Quantification of cannabisins from methanolic extracts of HHRs and HARs using UHPLC-QToF. Significantly higher amounts of cannabisins have been quantified in HHRs compared to HARs. Data are shown in µg/g. The values are displayed as the mean ± standard deviation of the biological triplicate (<span class="html-italic">n</span> = 3) and the technical triplicate (<span class="html-italic">n</span> = 3). Significant differences are indicated by asterisks: ***, <span class="html-italic">p</span> &lt; 0.001; ****, <span class="html-italic">p</span> &lt; 0.00001.</p>
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<p>Antifungal and antioxidant activities of HHR and HAR non-polar extracts: (<b>a</b>) Inhibition of <span class="html-italic">Saccharomyces cerevisiae</span> growth over time by 200 µg of HHR and HAR non-polar extracts prepared at 100 mg/mL in DMSO with DMSO as a negative control. (<b>b</b>) Radical Scavenging Activity shows as gallic acid equivalent, showing assessment of HHR and HAR non-polar extract antioxidant activities at 10 µg in reaction to ABTS solution. The values are displayed as the mean ± standard deviation of the technical replicate (<span class="html-italic">n</span> = 3).</p>
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<p>Establishment of hemp hairy root clones. (<b>a</b>) Selected hemp hairy root clone in solid culture media. (<b>b</b>) PCR analysis of HHR transformed with <span class="html-italic">R. rhizogenes</span> to verify the presence of <span class="html-italic">rol</span> B (T-DNA marker) and <span class="html-italic">Vir</span> G genes (<span class="html-italic">R. rhizogenes</span> marker). The DNA of <span class="html-italic">R. rhizogenes</span> was used as positive control (+) and water as negative control (−).</p>
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26 pages, 1451 KiB  
Review
Thraustochytrids: Evolution, Ultrastructure, Biotechnology, and Modeling
by Aleksei G. Menzorov, Daniil A. Iukhtanov, Ludmila G. Naumenko, Aleksandr V. Bobrovskikh, Ulyana S. Zubairova, Ksenia N. Morozova and Alexey V. Doroshkov
Int. J. Mol. Sci. 2024, 25(23), 13172; https://doi.org/10.3390/ijms252313172 (registering DOI) - 7 Dec 2024
Viewed by 112
Abstract
The thraustochytrids are a group of marine protists known for their significant ecological roles as decomposers and parasites as well as for their potential biotechnological applications, yet their evolutionary and structural diversity remains poorly understood. Our review critically examines the phylogeny of this [...] Read more.
The thraustochytrids are a group of marine protists known for their significant ecological roles as decomposers and parasites as well as for their potential biotechnological applications, yet their evolutionary and structural diversity remains poorly understood. Our review critically examines the phylogeny of this taxa, utilizing available up-to-date knowledge and their taxonomic classifications. Additionally, advanced imaging techniques, including electron microscopy, are employed to explore the ultrastructural characteristics of these organisms, revealing key features that contribute to their adaptive capabilities in varying marine environments. The integration of this knowledge with available omics data highlights the huge biotechnological potential of thraustochytrids, particularly in producing ω-3 fatty acids and other bioactive compounds. Our review underscores the importance of a systems biology approach in understanding thraustochytrids biology and highlights the urgent need for novel, accurate omics research to unlock their full biotechnological potential. Overall, this review aims to foster a deeper appreciation of thraustochytrids by synthesizing information on their evolution, ultrastructure, and practical applications, thereby providing a foundation for future studies in microbiology and biotechnology. Full article
(This article belongs to the Section Molecular Genetics and Genomics)
30 pages, 3535 KiB  
Review
Exploring Antimicrobial Compounds from Agri-Food Wastes for Sustainable Applications
by Mattia Di Maro, Luca Gargiulo, Giovanna Gomez d’Ayala and Donatella Duraccio
Int. J. Mol. Sci. 2024, 25(23), 13171; https://doi.org/10.3390/ijms252313171 (registering DOI) - 7 Dec 2024
Viewed by 133
Abstract
Transforming agri-food wastes into valuable products is crucial due to their significant environmental impact, when discarded, including energy consumption, water use, and carbon emissions. This review aims to explore the current research on the recovery of bioactive molecules with antimicrobial properties from agri-food [...] Read more.
Transforming agri-food wastes into valuable products is crucial due to their significant environmental impact, when discarded, including energy consumption, water use, and carbon emissions. This review aims to explore the current research on the recovery of bioactive molecules with antimicrobial properties from agri-food waste and by-products, and discusses future opportunities for promoting a circular economy in its production and processing. Mainly, antibacterial molecules extracted from agri-food wastes are phenolic compounds, essential oils, and saponins. Their extraction and antimicrobial activity against a wide spectrum of bacteria is analyzed in depth. Also, their possible mechanisms of activity are described and classified based on their effect on bacteria, such as the (i) alteration of the cell membrane, (ii) inhibition of energy metabolism and DNA synthesis, and iii) disruption of quorum sensing and biofilm formation. These bioactive molecules have a wide range of possible applications ranging from cosmetics to food packaging. However, despite their potential, the amount of wastes transformed into valuable compounds is very low, due to the high costs relating to their extraction, technical challenges in managing supply chain complexity, limited infrastructure, policy and regulatory barriers, and public perception. For these reasons, further research is needed to develop cost-effective, scalable technologies for biomass valorization. Full article
(This article belongs to the Special Issue Bioactive Materials with Antimicrobial Properties: 2nd Edition)
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<p>The application green principles to valorize agricultural waste within a circular economy framework. Reproduced from [<a href="#B11-ijms-25-13171" class="html-bibr">11</a>].</p>
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<p>Some examples of antimicrobial tests carried out using biomolecules obtained from agri-food waste valorization. (<b>1</b>) Antimicrobial activity of four grape seed extracts (<b>A</b>–<b>D</b>) against <span class="html-italic">S. aureus</span> (zone 1, 0.50 mg/mL extract; zone 2, 0.25 mg/mL; zone 3, 0.10 mg/mL; zone 4, 0.05 mg/mL; zone 5, negative control) (reproduced from [<a href="#B60-ijms-25-13171" class="html-bibr">60</a>]). (<b>2</b>) Inhibition halos obtained for <span class="html-italic">C. perfringens</span>, <span class="html-italic">C. botulinum</span> and <span class="html-italic">C. difficile</span> in the screening test using two different extracts of saffron petals (SPE A and SPE B) (reproduced from [<a href="#B70-ijms-25-13171" class="html-bibr">70</a>]) and (<b>3</b>) (<b>a</b>) MIC of ‘Maria Bruvele’ extract against <span class="html-italic">C. albicans</span> by the two-fold serial broth microdilution method; (<b>b</b>) MBC of the same extract against <span class="html-italic">P. aeruginosa</span> and <span class="html-italic">C. albicans</span> (reproduced from [<a href="#B65-ijms-25-13171" class="html-bibr">65</a>]).</p>
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<p>Schematic diagram of antimicrobial mechanisms exerted by biomolecules extracted from waste biomass.</p>
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<p><span class="html-italic">B. cereus</span> morphology observed by TEM. (<b>A</b>) <span class="html-italic">B. cereus</span> treated with 1/2 MIC. (<b>B</b>) <span class="html-italic">B. cereus</span> treated with MIC. (<b>C</b>) <span class="html-italic">B. cereus</span> treated with MBC. (<b>D</b>) positive control (<span class="html-italic">Cefixime</span>). (<b>E</b>) negative control (untreated <span class="html-italic">B. cereus</span>). Reprinted with permission from Ref. [<a href="#B126-ijms-25-13171" class="html-bibr">126</a>]. Copyright 2020 by Elsevier. For more clarity, a scale bar has been added under each image.</p>
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<p>Circular bioeconomy model in food packaging: integrating renewable resources in biorefineries to produce recyclable, eco-friendly materials. Reproduced from [<a href="#B127-ijms-25-13171" class="html-bibr">127</a>].</p>
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25 pages, 16929 KiB  
Article
Potential Antimicrobial and Cytotoxic Activity of Caralluma indica Seed Extract
by Shunmuga Vadivu Ramalingam, Senthil Bakthavatchalam, Karnan Ramachandran, Vasthi Gnanarani Soloman, Afrin Khan Ajmal, Mohammad Khalid Al-Sadoon and Ramachandran Vinayagam
Antibiotics 2024, 13(12), 1193; https://doi.org/10.3390/antibiotics13121193 (registering DOI) - 7 Dec 2024
Viewed by 185
Abstract
Background: Plant-derived phytochemicals are crucial in fighting bacterial infections and in cancer therapy. Objective: This study investigates the phytochemical composition of the ethanolic extract obtained from Caralluma indica (C. indica) seeds and assesses its antimicrobial, anticancer, and antioxidant activities. Results: GC-MS [...] Read more.
Background: Plant-derived phytochemicals are crucial in fighting bacterial infections and in cancer therapy. Objective: This study investigates the phytochemical composition of the ethanolic extract obtained from Caralluma indica (C. indica) seeds and assesses its antimicrobial, anticancer, and antioxidant activities. Results: GC-MS analysis found 30 phytochemicals in C. indica seeds, including 5 bioactive compounds that have been shown to have antioxidant, antimicrobial, and cytotoxicity properties, through in silico evaluation. Phytochemical screening of C. indica identified and measured the phenolic compounds, providing insight into its bioactive potential and therapeutic properties. C. indica exhibited robust antioxidant capacity (DPPH, ABTS, nitric oxide, and H2O2 radical scavenging) alongside potent antimicrobial activity against oral pathogen and cytotoxicity activity on a human oral squamous carcinoma cell line (OECM-1) (EC50 of 169.35 µg/mL) and yeast cell Saccharomyces cerevisiae (215.82 µg/mL), with a selective index of 1.27. The subminimum % MBC/MFC of C. indica significantly reduced biofilm formation against oral pathogens (p < 0.05). Molecular docking studies showed a strong correlation (r = 0.862) between antifungal and anticancer targets, suggesting that the antimicrobial agents in C. indica contribute to cancer prevention mechanisms. Conclusions: These findings propose C. indica seeds as promising candidates for combating oral pathogens, inhibiting biofilm formation, and reducing the risk of oral cancer progression. Full article
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<p>GC-MS chromatogram of the <span class="html-italic">C. indica</span> seed ethanolic extract.</p>
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<p>In vitro antioxidant activity of the <span class="html-italic">C. indica</span> seed ethanolic extract. (<b>a</b>) DPPH, (<b>b</b>) ABTS, (<b>c</b>) NO, and (<b>d</b>) H<sub>2</sub>O<sub>2</sub> radical scavenging; AA—ascorbic acid and CISEE—<span class="html-italic">C. indica</span> seed ethanolic extract.</p>
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<p>Antimicrobial activity of the <span class="html-italic">C. indica</span> seed ethanolic extract against oral-infection-causing pathogens. (<b>a</b>) <span class="html-italic">S. aureus</span> biofilm formation inhibitory activity of 25, 50, and 75% MBC concentrations (n = 3). (<b>b</b>) <span class="html-italic">C. albicans</span> biofilm formation inhibitory activity of 25, 50, and 75% MFC concentrations (n = 3). (<b>c</b>–<b>e</b>) Antimicrobial activity of <span class="html-italic">C. indica</span> seed ethanolic extract (50, 75, and 100 µg/mL) against oral pathogens (n = 3). (*) Statistically significant with the compared control (<span class="html-italic">p</span> &lt; 0.05); within each concentration, the different letters <sup>a,b,c</sup> indicate significance (<span class="html-italic">p</span> &lt; 0.05), and the same letters indicate non-significance (<span class="html-italic">p</span> &gt; 0.05), using a one-way ANOVA followed by Duncan’s multiple range test (DMRT); the significance level is <span class="html-italic">p</span> &lt; 0.05.</p>
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<p>In vitro cytotoxicity activity of the <span class="html-italic">C. indica</span> seed ethanolic extract. * significant difference from the control at <span class="html-italic">p</span> &lt; 0.05 using a one-way ANOVA followed by DMRT. (<b>a</b>) OECM-1 cell line. (<b>b</b>) Yeast <span class="html-italic">Saccharomyces cerevisiae</span> cell model.</p>
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<p>Morphology of normal and <span class="html-italic">C. indica</span> seed ethanolic extract-treated cells on OECM-1. (<b>a</b>) Control, (<b>b</b>–<b>f</b>) various concentrations of the CISEE extract, and (<b>g</b>) the standard as cisplatin.</p>
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<p>Morphological examination of the cytotoxicity (antiproliferative) activity of the <span class="html-italic">C. indica</span> seed ethanolic extract against yeast cells (the blue color arrow indicates a live cell (without staining) and the red color arrow indicates cell death (with blue color staining)). (<b>a</b>) Control and (<b>b</b>–<b>f</b>) various concentrations of the <span class="html-italic">C. indica</span> seed ethanolic extract.</p>
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<p>Molecular docking of the <span class="html-italic">C. indica</span> seed ethanolic extract phytochemicals against the antibacterial target of sortase A (PDB: 1T2W).</p>
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<p>Molecular docking of the <span class="html-italic">C. indica</span> seed ethanolic extract phytochemicals against the antibacterial target of sortase A (PDB: 1T2W).</p>
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<p>Molecular docking of <span class="html-italic">C. indica</span> seeds phytochemicals against the antifungal target of N-myristoyl transferase (PDB ID: 1NMT).</p>
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<p>Molecular docking of <span class="html-italic">C. indica</span> seed phytochemicals against the anticancer molecular target of DNMT1 (PDB ID: 4WXX).</p>
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<p>Molecular docking of <span class="html-italic">C. indica</span> seed phytochemicals against the anticancer molecular target of DNMT1 (PDB ID: 4WXX).</p>
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<p>Hierarchical tree clustering analysis between the targets (n = 5; <span class="html-italic">C. indica</span> seed bioactive compound binding affinity (kcal/mol) against targets on antimicrobial and anti-oral cancer).</p>
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<p>Graphical representation of the <span class="html-italic">C. indica</span> seed and the bioactive compound binding affinity score, which was similarly involved across all molecular targets for anti-oral microbial and anti-oral cancer.</p>
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<p>Molecular target homology modeling using phylogenetic tree analysis. (<b>a</b>) Antibacterial molecular target homology modeling. (<b>b</b>) Antifungal molecular target homology modeling. Red box indicates the targeted protein of micro orgaisms.</p>
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<p>Collection of <span class="html-italic">C. indica</span> seeds.</p>
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24 pages, 3827 KiB  
Article
Berries, Leaves, and Flowers of Six Hawthorn Species (Crataegus L.) as a Source of Compounds with Nutraceutical Potential
by Natalia Żurek, Michał Świeca and Ireneusz Tomasz Kapusta
Molecules 2024, 29(23), 5786; https://doi.org/10.3390/molecules29235786 (registering DOI) - 7 Dec 2024
Viewed by 134
Abstract
Designing new forms of food, food additives, and nutraceuticals is necessary due to the growing needs of consumers, as well as the inflammation of civilization diseases, the prevention and treatment of which can be significantly supported by dietary intervention. For this reason, this [...] Read more.
Designing new forms of food, food additives, and nutraceuticals is necessary due to the growing needs of consumers, as well as the inflammation of civilization diseases, the prevention and treatment of which can be significantly supported by dietary intervention. For this reason, this study aimed to obtain highly bioactive preparations in the form of powders from the fruits, leaves, and flowers of six species of hawthorn (Crataegus L.) using solid phase extraction (SPE). Ultra-performance liquid chromatography analysis (UPLC-PDA-MS/MS) showed a high concentration of phenolic compounds (in the range from 31.50 to 66.06 mg/g), including the highest concentration in hawthorn fruit preparations. Fruit preparations also showed the highest antioxidant activity (through scavenging of O2˙ and OH˙ radicals), antidiabetic activity (inhibition of α-amylase and α-glucosidase), and anticancer activity, mainly against colon cancer cells (Caco-2). At the same time, hawthorn flower preparations showed the highest biocompatibility against normal colon cells (CCD841CoN) and anti-inflammatory activity (trypsin inhibition). Correlation and principal component analysis (PCA) showed that the health-promoting potential was most influenced by the content of falavan-3-ols. The above findings provide a basis for the industrial use of the developed preparations, which is in line with the current trend in food technology related to the search for new sources of bioactive compounds and the design of highly bioactive food. Full article
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<p>pH value (<b>A</b>), equilibrium moisture content (<b>B</b>), and water solubility index (<b>C</b>) of hawthorn berries, leaves, and flower preparations. Results are expressed as mean (n = 3) ± SD. The values in the columns marked with different letters (lowercase letters, between species and morphological parts; uppercase letters, between means for morphological parts) indicate statistically significant differences (<span class="html-italic">p</span> &lt; 0.05). Explanations: C, <span class="html-italic">Crataegus</span>; 1–6, hawthorn species (see <a href="#sec2dot2-molecules-29-05786" class="html-sec">Section 2.2</a>. Plant material); B, berries; L, leaves; F, flowers.</p>
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<p>Total content of polyphenols (<b>A</b>), flavonoids (<b>B</b>), and proanthocyanidins (<b>C</b>) in hawthorn berries, leaves, and flower preparations. Results are expressed as mean (n = 3) ± SD. The values in the columns marked with different letters (lowercase letters, between species and morphological parts; uppercase letters, between means for morphological parts) indicate statistically significant differences (<span class="html-italic">p</span> &lt; 0.05). Explanations: C, <span class="html-italic">Crataegus</span>; 1–6, hawthorn species (see <a href="#sec2dot2-molecules-29-05786" class="html-sec">Section 2.2</a>. Plant material); B, berries; L, leaves; F, flowers.</p>
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<p>PCA analysis between studied hawthorn species, morphological parts, and analyzed parameters. Explanations: B, Berries; L, Leaves; F, Flowers; 1–6, hawthorn species (see <a href="#sec2dot2-molecules-29-05786" class="html-sec">Section 2.2</a>. Plant material).</p>
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<p>Pearson correlation illustrating the relationships between the studied variables for preparations obtained from hawthorn berries.</p>
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<p>Pearson correlation illustrating the relationships between the studied variables for preparations obtained from hawthorn leaves.</p>
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<p>Pearson correlation illustrating the relationships between the studied variables for preparations obtained from hawthorn flowers.</p>
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19 pages, 4014 KiB  
Review
Asteroid Saponins: A Review of Their Bioactivity and Selective Cytotoxicity
by Stuart J. Smith, Tianfang Wang and Scott F. Cummins
Mar. Drugs 2024, 22(12), 552; https://doi.org/10.3390/md22120552 (registering DOI) - 7 Dec 2024
Viewed by 141
Abstract
Saponins are a diverse class of secondary metabolites that are often reported to exhibit a variety of pharmacological applications. While research into the elucidation and application of plant and class Holothuroidea-derived saponins (i.e., sea cucumbers) is extensive, the class Asteroidea-derived saponins (i.e., seastars) [...] Read more.
Saponins are a diverse class of secondary metabolites that are often reported to exhibit a variety of pharmacological applications. While research into the elucidation and application of plant and class Holothuroidea-derived saponins (i.e., sea cucumbers) is extensive, the class Asteroidea-derived saponins (i.e., seastars) have been largely overlooked and primarily limited to elucidation. This review provides a comprehensive overview of the cytotoxic activities of asteroid-derived saponins against various cell cultures, for instance, mammalian erythrocytes, multiple microbial strains and cancer cell lines, including melanoma, breast, colon, and lung cancers. Highlighting the distinct structural variations in these saponins, this review examines their selective cytotoxicity and potency, with many demonstrating IC50 values in the low micromolar range. Specific compounds, such as asterosaponins and polyhydroxylated saponins, exhibit noteworthy effects, particularly against melanoma and lung carcinoma cells, while triterpenoid saponins were found to be highly cytotoxic to both erythrocytes and fungal cells. This review also addresses gaps in the research area, including the need for additional in vitro antimicrobial studies, in vivo studies, and further exploration of their mechanisms of action. By consolidating recent findings, we have shed light on the therapeutic potential of asteroid-derived steroidal saponins in developing novel antimicrobial and anticancer agents. Full article
(This article belongs to the Special Issue Biologically Active Compounds from Marine Invertebrates 2025)
26 pages, 1629 KiB  
Review
Quantitative Analysis of Caffeine in Roasted Coffee: A Comparison of Brewing Methods
by Iwona Mystkowska, Aleksandra Dmitrowicz and Monika Sijko-Szpańska
Appl. Sci. 2024, 14(23), 11395; https://doi.org/10.3390/app142311395 (registering DOI) - 6 Dec 2024
Viewed by 440
Abstract
Coffee is one of the most widely consumed beverages in the world due to its sensory and health benefits. The caffeine content, a bioactive compound of coffee, depends on many factors, including the brewing method, which is the subject of ongoing scientific research. [...] Read more.
Coffee is one of the most widely consumed beverages in the world due to its sensory and health benefits. The caffeine content, a bioactive compound of coffee, depends on many factors, including the brewing method, which is the subject of ongoing scientific research. In addition, various methods are used in studies to determine the caffeine content. However, it is worth noting that there is considerable variation in the individual analytical parameters within these methods. The aim of this study was to update the data on the effects of different brewing methods on the caffeine content of the brew and to present the current state of knowledge on techniques for the determination of this compound. A literature review was conducted, taking into account the latest studies in this field. The results showed that the caffeine content (mg/100 mL) of the brew prepared with the Cold Brew method was 48.50–179.95, Espresso—50.40–965.60, French Press—52.00–123.90, AeroPress—56.35–120.92, and Moka—128.00–539.90. These methods were characterized by different brewing parameters (time, water temperature and pressure, ratio of coffee to water), which differentiated the caffeine content. In addition, some methods were characterized by a wide range of caffeine content, suggesting that even minor variations in brewing method parameters may affect the content of this ingredient. High-performance liquid chromatography (HPLC) was the predominant method used. The detector wavelengths, along with other parameters of the HPLC method, such as gradient profiles and column temperatures, can affect the precision and accuracy of the analysis, and these differences can modify analyte retention and detection, leading to discrepancies in results. These results point to the need for studies that consider various brewing methods and a wide range of coffee types, including roast and origin, to accurately determine the effects of these factors on caffeine content determined by one precise method. Full article
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<p>The structure of caffeine and its main metabolites.</p>
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<p>Diagram of the search and selection process of articles included in the literature review. Source: based on Page et al. [<a href="#B28-applsci-14-11395" class="html-bibr">28</a>].</p>
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<p>Wavelengths used in the HPLC method to detect caffeine.</p>
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<p>Caffeine content in coffee by brewing method. (Results expressed in mg/100 mL to accurately reflect coffee consumption conditions. The interquartile range (IQR) was used to define a representative range for each brewing method).</p>
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18 pages, 3020 KiB  
Article
Metabolomics Characterization of Chemical Composition and Bioactivity of Highland Barley Monascus Tea Decoction Before and After Simulated Digestion In Vitro
by Haiyu Wu, Bin Dang, Wengang Zhang, Jie Zhang, Wancai Zheng, Jing Hao, Ping Ma and Xijuan Yang
Foods 2024, 13(23), 3950; https://doi.org/10.3390/foods13233950 - 6 Dec 2024
Viewed by 367
Abstract
A broadly targeted metabolomics approach based on UPLC-MS/MS was employed to investigate the changes in chemical composition and in vitro activity of highland barley Monascus tea decoction before and after simulated digestion. The characteristic metabolites of the tea decoction before and after in [...] Read more.
A broadly targeted metabolomics approach based on UPLC-MS/MS was employed to investigate the changes in chemical composition and in vitro activity of highland barley Monascus tea decoction before and after simulated digestion. The characteristic metabolites of the tea decoction before and after in vitro-simulated digestion were identified, and the in vitro antioxidant and enzyme inhibitory activities of the tea decoction were further analyzed. The study detected 1431 metabolites, including amino acids and their derivatives, alkaloids, organic acids, nucleotides and their derivatives, lipids, terpenoids, and phenolic acids. A total of 136 differential compounds were identified, primarily distributed in amino acids and their derivatives, alkaloids, organic acids, phenolics, and lipids. in vitro-simulated digestion significantly increased the content of amino acids, alkaloids, lipids, and phenolics in the tea. The differential metabolic compounds were primarily assigned to 20 metabolic pathways, mainly involving the metabolism of amino acids, nucleotides, carbohydrates, fatty acids, and other compounds. Additionally, after simulated digestion in vitro, the comprehensive antioxidant index (60.53%), α-glucosidase inhibitory activity (54.35%), and pancreatic lipase inhibitory activity (4.06%) was significantly improved. The highland barley Monascus tea decoction showed potential hypoglycemic and hypolipidemic efficacy. This study can provide a theoretical basis for the high-value utilization of highland barley and the development of healthy grain tea. Full article
(This article belongs to the Section Foodomics)
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<p>Distribution and variability of 1431 metabolites in highland barley Monascus tea decoction before and after simulated digestion in vitro. (<b>A</b>) Total ion flow plots of mixed samples and quality control (QC) samples-negative ion mode; (<b>B</b>) Total ion flow plots of mixed samples and quality control (QC) samples-positive ion mode; (<b>C</b>) MRM metabolite multi-peak detection plots-negative ion mode; (<b>D</b>) MRM metabolite multi-peak detection plots-positive ion mode; (<b>E</b>) Classifications and compositions of the 1431 metabolites; (<b>F</b>) Different classes of metabolites’ relative content; (<b>G</b>) principal component analysis of metabolites; (<b>H</b>) OPLS-DA score plot of metabolites.</p>
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<p>Clustering heat map of differential metabolites in highland barley Monascus tea decoction before and after simulated digestion in vitro.</p>
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<p>Classification and expression of different metabolites of highland barley Monascus tea decoction before and after simulated digestion in vitro. (<b>A</b>) Classification and composition of 136 differential metabolites; (<b>B</b>) the relative content of differentiated metabolites of different classes; (<b>C</b>) Up-regulated and down-regulated metabolite statistics; (<b>D</b>) Differential metabolites in the top 10 multiples of change.</p>
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<p>Changes in the content of five amino acids and their derivatives, five alkaloids, five organic acids, and 10 phenolic constituents in barley red tea broth before and after simulated digestion in vitro.</p>
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<p>KEGG annotation and enrichment results of differential metabolites of highland barley Monascus tea decoction before and after simulated digestion in vitro.</p>
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<p>In vitro activities of highland barley Monascus tea decoction before and after simulated digestion in vitro (<b>A</b>) DPPH radical scavenging ability, ABTS radical scavenging ability and FRAP reducing ability; (<b>B</b>) APC composite index (<b>C</b>) Inhibition of α-amylase and α-glucosidase activities; (<b>D</b>) Inhibition of lipase activity. Different lowercase letters in the figure represent significant differences between groups in different digestion stages of in vitro simulated digestion for the measured indexes.</p>
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17 pages, 1073 KiB  
Article
Bioactive Peptides in Greek Goat Colostrum: Relevance to Human Metabolism
by Maria Louiza Petre, Anna Nefeli Kontouli Pertesi, Olympia Eirini Boulioglou, Eleana Sarantidi, Artemis G. Korovesi, Athina Kozei, Angeliki I. Katsafadou, George T. Tsangaris, Antonia Trichopoulou and Athanasios K. Anagnostopoulos
Foods 2024, 13(23), 3949; https://doi.org/10.3390/foods13233949 - 6 Dec 2024
Viewed by 236
Abstract
Colostrum is essential for the survival and development of newborn mammals. This primary source of nourishment during the first days of infant life is rich in functional components conductive to the enhancement of neonate immunity and growth. Compared with mature milk, a higher [...] Read more.
Colostrum is essential for the survival and development of newborn mammals. This primary source of nourishment during the first days of infant life is rich in functional components conductive to the enhancement of neonate immunity and growth. Compared with mature milk, a higher protein and peptide content is observed in colostrum, whilst it is low in fat and carbohydrates. The functional properties of colostrum are closely linked to the release of bioactive peptides during the gastrointestinal digestion of colostrum proteins. Our study aimed to comprehensively analyze the whey proteome of colostrum from indigenous Greek goats and to examine the influence of bioactive peptides released during digestion on human metabolism. Colostrum and mature milk samples from healthy ewes were subjected to nanoLC-MS/MS analysis, revealing differentially expressed proteins. These proteins were functionally characterized and subjected to in silico digestion. Using machine learning models, we classified the peptide functional groups, while molecular docking assessed the binding affinity of the proposed angiotensin-converting enzyme (ACE)- and dipeptidyl peptidase IV (DPPIV)-inhibitory peptides to their target molecules. A total of 898 proteins were identified in colostrum, 40 of which were overexpressed compared with mature milk. The enzymatic cleavage of upregulated proteins by key gastrointestinal tract proteases and the downstream analysis of peptide sequences identified 117 peptides predicted (with >80% confidence) to impact metabolism, primarily through modulation of the renin–angiotensin system, insulin secretion, and redox pathways. This work advances our understanding of dietary bioactive peptides and their relevance to human metabolism, highlighting the potential health benefits of colostrum consumption. Full article
(This article belongs to the Section Food Physics and (Bio)Chemistry)
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