Sustainable Extraction Protocols for the Recovery of Bioactive Compounds from By-Products of Pomegranate Fruit Processing.

This study investigates sustainable extraction protocols for the recovery of bioactive compounds from by-products of various pomegranate (Punica granatum L.) cultivars, including Acco, Hicaz, Jolly Red, Parfianka, Valenciana, and Wonderful, generated during the industrial processing of the fruits. Advanced extraction technologies, including ultrasounds, microwaves, and hydrodynamic cavitation, have been compared to conventional extraction procedures and utilized to enhance extraction efficiency while also minimizing environmental impact. Water-based extraction methods have been utilized to promote the development of sustainable and eco-friendly processes. The comparison between conventional extractions and ultrasound-assisted extractions (UAEs) and microwave-assisted extractions (MAEs) demonstrated notable improvements in extraction yields, particularly for ellagitannins (punicalins, punicalagins, and ellagic acid) and total polyphenols, with increases ranging from about 45 to 200%. However, the increases directly comparing UAEs to MAEs ranged from about 4 to 6%. This indicates that while both UAEs and MAEs offer notable improvements over conventional extractions, the differences in extraction efficiency between the two advanced methods were relatively modest. These advancements were observed across various pomegranate cultivars, highlighting the versatility and effectiveness of these methods. Notably, hydrodynamic cavitation-based extractions (HC) emerged as particularly promising, consistently yielding the highest levels of bioactive compounds (ellagitannins and total polyphenols), especially when operated at higher frequencies. Compared to conventional extractions, HC exhibited substantial increases in extraction yields for Wonderful pomegranate by-products, surpassing the efficiency of both UAEs and MAEs (approximately 45 and 57% for UAE and MAE, respectively, versus about 80% for HC). Among these advanced techniques, HC has emerged as particularly promising, yielding the most favorable results and leading to significant improvements in the yield of bioactive compounds. When directly compared to UAEs and MAEs, HC increased extraction yields by over 20%. Furthermore, HC allowed for shorter extraction times. The Wonderful cultivar consistently exhibited the highest levels of ellagitannins and the highest total polyphenol content among all types of extraction procedures used, whether conventional or advanced. This highlights the great potential of the Wonderful cultivar in terms of bioactive compound extraction and underscores its significance in research and applications related to pomegranate processing and utilization. This study suggests that the implementation of these advanced technologies into extraction processes represents a significant advancement in the field, offering a promising avenue for the development of efficient and environmentally friendly extraction methods for obtaining valuable bioactive compounds from pomegranate processing by-products.

Green Synthesis of Silver Nanoparticles Mediated by Punica granatum Peel Waste: An Effective Additive for Natural Rubber Latex Nanofibers Enhancement.

Pomegranate waste poses an environmental challenge in Arequipa. Simultaneously, interest in sustainable materials like natural rubber latex (NRL) is growing, with Peruvian communities offering a promising source. This study explores the green synthesis of silver nanoparticles (AgNPs) using pomegranate peel extract and their incorporation into NRL nanofibers for enhanced functionalities. An eco-friendly process utilized silver nitrate and pomegranate peel extract as a reducing and capping agent to synthesize AgNPs. The resulting AgNPs and NRL/AgNPs nanofibers were characterized using imaging and spectroscopic techniques such as UV-vis, TGA, FTIR, XRD, Raman, SEM, and DLS. Green-synthesized AgNPs were spherical and crystalline, with an average diameter of 59 nm. They showed activity against K. pneumoniae, E. coli, B. cereus, and S. aureus (IC50: 51.32, 4.87, 27.72, and 69.72 µg/mL, respectively). NRL and NRL/AgNPs nanofibers (300-373 nm diameter) were successfully fabricated. The composite nanofibers exhibited antibacterial activity against K. pneumoniae and B. cereus. This study presents a sustainable approach by utilizing pomegranate waste for AgNP synthesis and NRL sourced from Peruvian communities. Integrating AgNPs into NRL nanofibers produced composites with antimicrobial properties. This work has potential applications in smart textiles, biomedical textiles, and filtration materials where sustainability and antimicrobial functionality are crucial.

Comparative Study of Callistemon citrinus (Bottlebrush) and Punica granatum (Pomegranate) Extracts for Sustainable Synthesis of Silver Nanoparticles and Their Oral Antimicrobial Efficacy.

A comparative study was applied to investigate the potential of Callistemon citrinus (bottlebrush) flower extract (BBE) and Punica granatum (pomegranate) peel extracts (PPE) for the sustainable synthesis of the silver nanoparticles, Ag-BBE and Ag-PPE, respectively. The synthesis process of Ag NPs using the selected extracts was applied under optimized conditions. Hence, the effect of the selected plant's type on the different characteristics of the synthesized green Ag NPs was investigated. The UV-Vis spectroscopy revealed the presence of the characteristic silver peaks at 419 and 433 nm of the Ag-BBE and Ag-PPE, respectively. The XRD spectra reported the fcc phase formation of Ag NPs. The TEM results highlighted the morphological features of the synthesized Ag NPs. with a size range of 20-70 nm, and with 10-30 nm for Ag-BBE and Ag-PPE, correspondingly. The Raman spectra revealed characteristic silver bands in the Ag-PPE and reflected some bands related to the natural extract in the Ag-BBE sample. The antimicrobial activity and statistical analysis investigation were conducted against four selected oral pathogens (Staphylococcus aureus (SA), Candida albicans (CA), Staphylococcus epidermidis (S. epi), and Enterococcus faecalis (EF)). Both tested extracts, BBE, and PPE, revealed potential effectivity as reducing and capping agents for Ag NP green synthesis. However, the synthesized NPs demonstrated different features, depending on the used extract, reflecting the influence of the plant's biomolecules on the nanoparticles' properties.

Polyphenol-Rich Extracts from Agroindustrial Waste and Byproducts: Results and Perspectives According to the Green Chemistry and Circular Economy.

Polyphenols are natural secondary metabolites found in plants endowed with multiple biological activities (antioxidant, anti-inflammatory, antimicrobial, cardioprotective, and anticancer). In view of these properties, they find many applications and are used as active ingredients in nutraceutical, food, pharmaceutical, and cosmetic formulations. In accordance with green chemistry and circular economy strategies, they can also be recovered from agroindustrial waste and reused in various sectors, promoting sustainable processes. This review described structural characteristics, methods for extraction, biological properties, and applications of polyphenolic extracts obtained from two selected plant materials of the Mediterranean area as olive (Olea europaea L.) and pomegranate (Punica granatum L.) based on recent literature, highlighting future research perspectives.

Citric acid cross-linked pomegranate peel extract-loaded pH-responsive ß-cyclodextrin/carboxymethyl tapioca starch hydrogel film for diabetic wound healing.

Pomegranate peel extract (PPE) hydrogel films filled with citric acid (CA) and ß-cyclodextrin-carboxymethyl tapioca starch (CMS) were designed mainly to prevent wound infections and speed up the healing process. FTIR and NMR studies corroborated the carboxymethylation of neat tapioca starch (NS). CMS exhibited superior swelling behavior than NS. The amount of CA and ß-CD controlled the physicochemical parameters of developed PPE/CA/ß-CD/CMS films. Optimized film (OF) exhibited acceptable swellability, wound fluid absorptivity, water vapor transmission rate, water contact angle, and mechanical properties. Biodegradable, biocompatible, and antibacterial films exhibited pH dependence in the release of ellagic acid for up to 24 h. In mice model, PPE/CA/ß-CD/CMS hydrogel film treatment showed promising wound healing effects, including increased collagen deposition, reduced inflammation, activation of the Wingless-related integration site (wnt) pathway leading to cell division, proliferation, and migration to the wound site. The expression of the WNT3A gene did not show any significant differences among all the studied groups. Developed PPE-loaded CA/ß-CD/CMS film promoted wound healing by epithelialization, granulation tissue thickness, collagen deposition, and angiogenesis, hence could be recommended as a biodegradable and antibacterial hydrogel platform to improve the cell proliferation during the healing of diabetic wounds.

The effects of pomegranate (Punica granatum) on nonalcoholic fatty liver disease: A systematic review of in vivo interventional studies.

Nonalcoholic fatty liver disease (NAFLD) is a disorder in which excess fat accumulates in hepatocytes and can lead to serious complications. Oxidative stress is one of the leading causes of NAFLD. Pomegranates are considered antioxidant-rich fruit. This systematic review study was aimed to investigate the impact of pomegranate on NAFLD. PubMed, Scopus, and Google Scholar databases/search engines (from inception up to July 2023) were searched for interventional studies (human and animal) that examined the effects of supplementation with different parts of pomegranate including fruits, peels, seeds, or flower on NAFLD outcomes. A total of 222 articles were retrieved following the initial search. After excluding duplicates, the title and abstract of 114 articles were screened. Afterward, irrelevant articles were removed and the full texts of the remaining 27 articles were reviewed. Eventually, 19 articles (16 animal and three human interventional studies) that met the inclusion criteria, published between 2009 and 2023, were included in this systematic review. Our study indicates the potential beneficial effects of different parts of pomegranate on the improvement of NAFLD. However, given that the majority of the included articles were animal studies, further investigations in the form of human clinical trials are warranted to suggest a clinical indication of such interventions.

Anthocyanins from pomegranate peel (Punica granatum), chili pepper fruit (Capsicum annuum), and bougainvillea flowers (Bougainvillea spectabilis) with multiple biofunctions: Antibacterial, antioxidant, and anticancer.

Background: Natural colorants, including natural pigments, e.g., anthocyanins, carotenoids, and chlorophylls, in novel and attractive food matrixes have become a popular trend. They impart favorite colors to food products and provide significant therapeutic effects. This study is aimed at extracting and identifying some natural pigments from different plant sources and evaluating their ability as antibacterial, antioxidant, and anticancer activities. Methods: The anthocyanin-rich extract (ARE) is derived from three natural plant sources: pomegranate peel (Punica granatum), chili pepper fruit (Capsicum annuum), and Bougainvillea flowers. Bougainvillea spectabilis are analyzed for biochemical composition, as well as antioxidant, antibacterial, and anticancer activity, HPLC, DPPH, FRAP, disc diffusion assay, MIC, MTT, VEGFR-2, and caspase-9 assays. Results: All three extracts had varying total phenolic contents, ranging from 14 to 466 mg GAE/g extract, where Punica granatum was the highest (466 mg GAE/g extract), followed by Bougainvillea spectabilis (180 mg GAE/g extract), and then Capsicum annuum (14 mg GAE/g extract). The antioxidant activity rose steadily with raising concentration. The ARE of pomegranate peels recorded highest value, followed by Bougainvillea flowers and chili pepper fruit. The MTT assay revealed an inhibitory action of the tested extracts on the proliferation of HCT-116, MCF-7, and HepG2 in a concentration-based manner. Gene expression of caspase-9 transcripts was considerably multiplied by the application of ARE of pomegranate peels. All the tested extracts inhibited VEGFR-2, and the inhibition (%) expanded gradually with increasing concentrations, achieving the highest value (80 %) at 10 µg/mL. The ARE of pomegranate peels scored highest antibacterial activity, followed by ARE of chili pepper fruit and Bougainvillea flowers. The inhibition zone diameter escalated gradually with rising concentrations of the tested samples. Conclusion: The AREs of the three studied plant sources can be used as multifunctional products with antioxidant, anticancer, and antibacterial activities that are natural, safe, and cheap.

A Genome-Wide Analysis of the BAM Gene Family and Identification of the Cold-Responsive Genes in Pomegranate (Punica granatum L.).

Beta-amylases (BAMs, EC 3.2.1.2), belonging to a multigene family, play a pivotal role in starch breakdown and are also involved in hormonal and stress responses, notably to cold stress. Pomegranate trees (Punica granatum L.) are adapted to warm climates and are sensitive to cold temperatures. In this study, we analyzed eight PgBAM genes from the pomegranate genome dataset. These members unevenly distributed across chromosomes and were categorized into four groups based on their orthologous members. The motif composition was highly consistent among most members. In contrast, exon numbers and arrangements were conserved within groups or subgroups, whereas significant diversity was observed between different groups. A syntenic analysis revealed that three PgBAM members (PgBAM1/4/5) showed a total of 11 syntenic relationships with the BAM members from Arabidopsis, kiwifruit, and Chinese white pear, respectively. Promoter binding motif prediction suggested potential roles for PgBAMs' genes in light, stress, hormones, and development signaling. Gene expression indicated that PgBAM4 was predominantly expressed in leaves, PgBAM7 in flowers, and PgBAM8 in roots and leaves and during fruit ripening, particularly in pericarp development. A transcriptome analysis identified the starch and sucrose metabolism pathway (map00500) as a key factor in the cold stress response of cold-sensitive cultivar 'Tunisia' seedlings. PgBAM4 exhibited remarkable expression and was closely associated with the cold-responsive BAM genes, characterized by a closer phylogenetic relationship, conserved catalytic residues, and similar secondary and tertiary structures. Moreover, the differences in soluble sugar levels and PgBAM4 expression were closely associated with the varying cold stress resistance observed between 'Tunisia' and 'Sanbai' seedlings. Furthermore, yeast one-hybrid assays confirmed that PgCBF7, a critical transcription factor for enhancing freezing tolerance, binds to the promoter region of PgBAM4. Our findings provide a systematic overview of the PgBAM gene family and shed new light on the regulatory mechanisms underlying cold stress tolerance in pomegranate.

Development of a New Extraction Method for Pomegranate and Metabolite Profiling by a LC-MS and 1H NMR Combined Approach.

The pomegranate (Punica granatum L.) market has steadily grown due to the increasing demand of health-conscious consumers of products with health-promoting effects. Recently, a growing interest in developing ecological and environmentally friendly extraction methods has led to investigating extraction procedures to obtain a higher extraction yield using a lower amount of solvents and energy. Herein, a new extraction procedure was developed to obtain an enriched pomegranate food supplement by using the peels of pomegranate, cultivar "Dente di Cavallo" and its juice. The extraction was performed through a non-conventional extraction technique like SLDE-Naviglio using ethanol and pomegranate juice as a solvent, and peels as a matrix. The extract was analysed by a combined approach based on LCESI/QExactive/MS/MS and NMR analysis, and its chemical profile was compared with those of pomegranate juice and of the extract obtained from peels by SLDE-Naviglio by using ethanol:H2O. The LC-MS analysis highlighted the presence of hydrolysable tannins, flavonoids, ellagic acid and phenol glucoside derivatives, while 1H NMR analysis completed the profile by detecting the primary metabolites. The LC-MS and 1H NMR analysis indicated that the extract obtained by SLDE-Naviglio using ethanol and pomegranate juice was enriched in the bioactives as confirmed by the highest phenolic, tannin and flavonoid content.

Pomegranate Rhizosphere Microbial Diversity Revealed by Metagenomics: Toward Organic Farming, Plant Growth Promotion and Biocontrol?

Food production must undergo systems change to meet the sustainable development goals (SDGs). For example, organic farming can be empowered by soil microorganisms with plant growth promotion (PGP) and biocontrol features. In this context, there have been limited studies on pomegranate. We investigated microbial diversity in rhizosphere of the pomegranate "Bhagwa" variety and its potential role in PGP and biocontrol. Both bulk and rhizosphere soil samples were analyzed for their physicochemical properties. Whole metagenome sequencing was conducted using the Illumina NovaSeq6000 platform. Surprisingly, we found that bulk and rhizosphere soil samples had comparable microbial diversity. Metagenome sequencing revealed the abundance of Streptomyces indicus, Bradyrhizobium kalamazoonesis, and Pseudomonas cellulosum in the rhizosphere that are reported here for the first time in agricultural literature. Pathway prediction analysis using KEGG (Kyoto Encyclopedia for Genes and Genomes) and COG (clusters of orthologous genes) databases identified metabolic pathways associated with biocontrol properties against pathogens. We confirmed the metagenome data in vitro, which demonstrated their PGP potential and antimicrobial properties. For instance, S. indicus produced high concentration of indole-3-acetic acid, a PGP phytohormone, that can stimulate plant growth. In addition, an antimicrobial susceptibility assay suggested that bacterial extracts displayed activity against Xanthomonas, a primary pathogen causing the pomegranate wilt disease. In conclusion, this study suggests that S. indicus, B. kalamazoonesis, and P. cellulosum can potentially be PGP and biocontrol agents that may contribute to increased crop productivity in pomegranate cultivation. These agents and their combinations warrant future research with an eye on SDGs and so as to enable and innovate organic farming and pomegranate agricultural practices.

Green and sustainable smart wooden system integrated with cellulose nanowhiskers-supported polyvinyl alcohol and anthocyanin biomolecules to monitor food freshness.

Smart packaging materials have been used to protect human health from environmental hazards by sending real-time colorimetric signals for changes in the food packaging environment. However, the colorimetric material sensors use synthetic sensor dyes, which are toxic, expensive, non-biodegradable, and difficult to prepare. Herein, a simple strategy is presented for the development of an environmentally-friendly halochromic wood able to change color upon exposure to spoilage of food. A combination of anthocyanin (Ac)/aluminum (Al) mordant (Ac/Al) nanoparticles and cellulose nanowhiskers (CNW)-reinforced polyvinyl alcohol (PVA) was infiltrated into a delignified wood to produce a translucent wood with halochromic properties. CNW were employed as reinforcement agent to improve the mechanical performance of PVA. Additionally, CNW function as a dispersing agent to prevent agglomeration of Ac/Al nanoparticles. The diameters of CNW are in the range of 12-19 nm, whereas Ac/Al particles showed diameters of 9-22 nm. The smart wood changed color from purplish to colorless when exposed to food spoilage. A hypsochromic change from 539 nm to 370 nm was shown by the anthocyanin receptor when the spoilage level of food increased. This could be attributed to the pH-driven molecular switching of anthocyanin, leading to charge delocalization.

Pomegranate (Punica granatum L.) phytochemicals target the components of metabolic syndrome.

Pomegranate (Punica granatum L.) is a multipurpose dietary and medicinal plant known for its ability to promote various health benefits. Metabolic syndrome (MetS) is a complex metabolic disorder driving health and socioeconomic challenges worldwide. It may be characterized by insulin resistance, abdominal obesity, hypertension, and dyslipidemia. This study aims to conduct a review of pomegranate's effects on MetS parameters using a mechanistic approach relying on pre-clinical studies. The peel, juice, roots, bark, seeds, flowers, and leaves of the fruit present several bioactive compounds that are related mainly to anti-inflammatory and antioxidant activities as well as cardioprotective, antidiabetic, and antiobesity effects. The use of the juice extract can work as a potent inhibitor of angiotensin-converting enzyme activities, consequently regulating blood pressure. The major bioactive compounds found within the fruit are phenolic compounds (hydrolysable tannins and flavonoids) and fatty acids. Alkaloids, punicalagin, ellagitannins, ellagic acid, anthocyanins, tannins, flavonoids, luteolin, and punicic acid are also present. The antihyperglycemia, antihyperlipidemia, and weight loss promoting effects are likely related to the anti-inflammatory and antioxidant effects. When considering clinical application, pomegranate extracts are found to be frequently well-tolerated, further supporting its efficacy as a treatment modality. We suggest that pomegranate fruit, extract, or processed products can be used to counteract MetS-related risk factors. This review represents an important step towards exploring potential avenues for further research in this area.

The power of Punica granatum: A natural remedy for oxidative stress and inflammation; a narrative review.

ETHNOPHARMACOLOGICAL RELEVANCE: Pomegranate 'Punica granatum' offers multiple health benefits, including managing hypertension, dyslipidemia, hyperglycemia, insulin resistance, and enhancing wound healing and infection resistance, thanks to its potent antioxidant and anti-inflammatory properties. It has been symbolized by life, health, femininity, fecundity, and spirituality. AIM OF THE STUDY: Although laboratory and animal studies have been conducted on the healing effects of pomegranate, there needs to be a comprehensive review on its anti-oxidative and anti-inflammatory effects in chronic disorders. We aim to provide a comprehensive review of these effects based on in-vitro, in-vivo, and clinical studies conducted in managing various disorders. MATERIALS AND METHODS: A comprehensive search of in-vitro, in-vivo, and clinical findings of pomegranate and its derivatives focusing on the highly qualified original studies and systematic reviews are carried out in valid international web databases, including Web of Science, PubMed, Scopus, and Cochrane Library. RESULTS: Relevant studies have demonstrated that pomegranate and its derivatives can modulate the expression and activity of several genes, enzymes, and receptors through influencing oxidative stress and inflammation pathways. Different parts of pomegranate; roots, bark, blossoms, fruits, and leaves contain various bioactive compounds, such as polyphenols, flavonoids, anthocyanins, and ellagitannins, that have preventive and therapeutic effects against many disorders such as cardiovascular diseases, diabetes, neurological diseases, and cancers without any serious adverse effects. CONCLUSIONS: Most recent scientific evidence indicates that all parts of the pomegranate can be helpful in treating a wide range of chronic disorders due to its anti-oxidative and anti-inflammatory activities. Since the safety of pomegranate fruit, juice, and extracts is established, further investigations can be designed by targeting its active antioxidant and anti-inflammatory constituents to discover new drugs.

Effects of natural waste on in vitro oxidative DNA damage.

In this study, the effects of natural waste products such as extracts from C. coggygria leaves and Punica granatum L. peels were investigated against oxidative DNA damage induced by Fenton reaction. 8-OH-2'dG as a general marker of DNA damage on thymus DNA, and the bioactive compounds of extracts were measured by LC-MS/MS. Our results had shown that ethanol extracts of C. coggygria leaf and Punica granatum L. peel had a protective effect on oxidative damaged DNA. It was determined that the bioactive compounds of C. coggygria leaves (gallic acid, protocatechuic acid, myricetin, syringic acid and ethyl gallate as a major compounds) and Punica granatum L. peel (ellagic acid, abscisic acid, ethyl gallate, phlorizin, gallic acid, myricetin as major compounds) may have an important role in the protective effect against oxidative DNA damage. Therefore, Cotinus coggygria leaves and Punica granatum L. peel may have potential use in medicine or cosmetic fields.

Punica granatum L. (Pomegranate) Extracts and Their Effects on Healthy and Diseased Skin.

The aim of this review is to provide a summary of the botany, phytochemistry and dermatological effects of Punica granatum (PG), with special emphasis on therapeutic mechanisms in various skin conditions. PG peel contains the highest levels of chemical compounds. Due to the high abundance of polyphenolic compounds, including phenolic acids, anthocyanins and flavonoids, exhibiting strong antioxidant properties, PG peel possesses significant health-promoting effects. Up until now, different parts of PG in the form of various extracts, fixed seed oil or individual active compounds have been investigated for various effects on skin conditions in in vitro and in vivo studies, such as antioxidant, anti-inflammatory, antimicrobial, chemoprotective and antiaging effects, as well as positive effects on striae distensae, skin repair mechanisms, erythema, pigmentation and psoriasis. Therefore, formulations containing PG active compounds have been used for skincare of diseased and healthy skin. Only a few effects have been confirmed on human subjects. Based on encouraging results obtained in in vitro and animal studies about the numerous substantial dermatological effects of PG active compounds, future perspectives should incorporate more in vivo investigations in human volunteers. This approach can aid in identifying the optimal concentrations and formulations that would be most efficacious in addressing specific skin conditions.

Development of toxic gas sensor from anthocyanin-embedded polycaprolactone-co-polylactic acid nanofibrous mat.

The colorless ammonia gas has been a significant intermediate in the industrial sector. However, prolonged exposure to ammonia causes harmful effects to organs or even death. Herein, an environmentally friendly solid-state ammonia sensor was developed utilizing colorimetric polycaprolactone-co-polylactic acid nanofibrous membrane. Pomegranate (Punica granatum L.) peel contains anthocyanin (ACN) as a naturally occurring spectroscopic probe. A mordant (potassium aluminum sulfate) is used to immobilize the anthocyanin direct dyestuff inside nanofibers, generating mordant/anthocyanin (M/ACN) coordinated complex nanoparticles. When exposed to ammonia, the color change of anthocyanin-encapsulated polycaprolactone-co-polylactic acid nanofibrous membrane from purple to transparent was examined by absorbance spectra and CIE Lab color parameters. With a quick colorimetric shift, the polycaprolactone-co-polylactic acid fabric exhibits a detection limit of 5-150 mg/L. The absorbance spectra showed a hypsochromic shift when exposed to ammonia, displaying an absorption shift from 559 nm to 391 nm with an isosbestic point of 448 nm. Scanning electron microscopy (SEM) images revealed that the polycaprolactone-co-polylactic acid nanofibers had a diameter of 75-125 nm, whereas transmission electron microscopy (TEM) images revealed that M/ACN nanoparticles exhibited diameters of 10-20 nm.

Thermal processing of pomegranate seed oils underscores their antioxidant stability and nutritional value: Comparison of pomegranate seed oil with sesame seed oil.

In the present study, the oxidative stability and antioxidant activity of seed oils were investigated in three Iranian pomegranate cultivars, Shirin Khafr, Torsh Sabz, and Rabab, along with the sesame (Sesamum indicume L. cv Dezful) seed oil. Punicic acid was the primary fatty acid in the pomegranate seed oils, with contents ranging from 75.5 to 80.9% (w/w). The tocopherol levels in pomegranate seed oils ranged from 1439 to 2053 mg/kg, whereas the phenolics ranged from 130 to 199.3 mg/kg, respectively. Comparatively, in the seed oil of sesame "Dezful," these substances' contents were 1053 and 79 mg/kg, respectively. Contrary to common perception, the seed oil of the three pomegranate cultivars cultivated in Iran had high oxidative stability and antioxidative activity during the 32 h of thermal processing at 170°C. The oxidation stability assayed by peroxide value, p-anisidine value, and TOTOX index revealed that the pomegranate seed oils had a much higher resistance to the oxidation process than the sesame oil. The content of tocopherols increased during thermal processing due to the regeneration phenomenon. Tocopherols are not always free and may form a matrix with themselves or other compounds. Changes in the antioxidant activity during the thermal processing assessed by DPPH free radical scavenging power and by the FRAP test were consistent with those for the antioxidants. Therefore, these oils can be added to other edible oils as a natural antioxidant to improve their oxidative stability.

Unlocking the Immunomodulatory Potential of Rosmarinic Acid Isolated from Punica granatum L. Using Bioactivity-Guided Approach: In Silico, In Vitro, and In Vivo Approaches.

BACKGROUND: Punica granatum L. is well-known for its multifaceted therapeutic potential, including anti-inflammatory and immunomodulatory activities. AIM: This study aimed to characterize an immunomodulatory compound isolated from Punica granatum L. using a bioactivity-guided approach. METHODS: Chromatographic techniques were adopted for isolation and purification of secondary metabolites. In silico, in vitro, and in vivo methods were performed to characterize the therapeutic potential of the isolated compound. RESULTS: Using preparative thin-layer chromatography, rosmarinic acid was isolated from F4 (column chromatography product obtained from a butanolic fraction of the extract). The impact of rosmarinic acid was assessed in rats using the neutrophil adhesion test, DTH response, and phagocytic index. In immunized rats, rosmarinic acid demonstrated significant immunomodulatory potential. Computational experiments, like molecular docking and molecular dynamics, were also conducted against two targeted receptors, Cereblon (PDB ID: 8AOQ) and human CD22 (PDB ID: 5VKM). Computational studies suggested that an increase in phagocytic index by rosmarinic acid could be attributed to inhibiting Cereblon and CD22. Pharmacokinetics and toxicity prediction also suggested the drug-likeness of rosmarinic acid. CONCLUSION: Rosmarinic acid is a potential candidate, but extensive research needs to be done to translate this molecule from bench to bedside.

Pomegranate extract on eroded dentin: antioxidant action, bond strength and morphology of the adhesive interface after aging.

Objectives: This study aimed to evaluate the effect of pomegranate solution (Punica granatum) on eroded dentin through antioxidant action, shear bond strength (SBS) and interface morphology. Materials and Methods: The 10% pomegranate peel extract was prepared by the lyophilization method. Punicalagin polyphenol was confirmed by high-performance liquid chromatography. Antioxidant activity was evaluated by capturing the 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical. For the SBS, 48 dentin fragments were divided into sound or eroded, and subdivided according to the pretreatment (n = 12): water or P. granatum. The surfaces were restored with self-etch adhesive and a bulk-fill resin (Ecosite; DMG). The SBS was done immediately (24 hours) and after thermal cycling + water storage (12 months). For scanning electron microscopy, 48 dentin fragments (24 sound and 24 eroded) received the same treatments as for SBS (n = 6), and they were analyzed after 24 hours and 12 months. Results: The P. granatum had antioxidant action similar (p = 0.246) to the phenolic standard antioxidants. After 24 hours, eroded dentin had lower SBS than sound dentin (p < 0.001), regardless of the pretreatment. After 12 months, P. granatum maintained the SBS of sound dentin (13.46 ± 3.42 MPa) and eroded dentin (10.96 ± 1.90 MPa) statistically similar. The lowest values were found on eroded dentin treated with water (5.75 ± 1.65 MPa) (p < 0.001). P. granatum on eroded dentin caused peritubular demineralization and hybrid layer with resin tags. Conclusions: The pomegranate extract had antioxidant action and preserved the adhesive interface of the eroded dentin.

First Report of Colletotrichum siamense Causing Fruit Anthracnose on Pomegranate (Punica granatum) in China.

As a native crop in central Asia, pomegranate (Punica granatum L.) has been cultivated in China for more than 2000 years (Yuan et al. 2007). In August 2022, typical symptoms of anthracnose were observed on pomegranate fruitlets in the main cultivation area (34°22'36″N, 109°15'58″E) in Shaanxi Province, China. The disease incidence was approximately 10 to 15% in the field. The initial symptoms were slightly small, light, dark, sunken lesions with irregular, circular shapes. As the disease progressed, the necrotic lesions gradually expanded and merged, eventually leading to the abscission of fruits (Figure 1, A). The symptomatic lesion samples of the pomegranate were sterilized for 1.5 min in 75% ethanol and 2 min in 1% NaClO and rinsed for 2 min in sterile water three times. The sterilized samples were dried on sterile filter paper and placed on potato dextrose agar (PDA) media at 25 ℃ for 5 days. The mycelia of the isolate were white, cottony, and diffuse (Figure 1, B and C). The conidia were single-celled, smooth, aseptate, and cylindrical with slightly rounded ends, measured 13.5 to 17.5 µm long and 3.5 to 6.5 µm wide (mean 16.0 × 4.5 µm, n = 50) (Figure 1, D). These morphological characteristics were identical to those of Colletotrichum siamense (Weir et al. 2012; Cannon et al. 2012; Zhuo et al. 2022). For accurate molecular characterization of the fungus, genomic DNA was extracted from the hyphae of the two isolates using microorganism lysis buffer (Takara, Japan). The internal transcribed spacer (ITS), calmodulin (CAL), actin (ACT), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), and beta-tubulin (TUB2) regions were amplified and sequenced. All the sequences were submitted to GenBank with accession numbers OQ832556 (ITS), OQ848589 (GAPDH), OQ848590 (ACT), OQ848591 (CAL), and OQ986593 (TUB2). The isolates showed 99 to 100% identity with sequences of Colletotrichum siamense (100% with GAPDH, ACT, CAL, and TUB2; 99.81% with ITS). The morphology of the strain was studied, and multilocus (ITS, GAPDH, ACT, CAL, and TUB2) phylogenetic analysis was performed (Figure 2). Therefore, the causal pathogen was identified as C. siamense based on the results of morphological and molecular analyses. Pathogenicity assays were performed on pomegranate (cv. Lishanhong) fruits. A conidial suspension (1×106 conidia/mL) was sprayed onto 10 unwounded fruits to inoculate them as infected samples, and the controls were inoculated with a sterile water suspension. All the samples were maintained in an artificial climate box at 25 ± 2 ℃ with 70% relative humidity, and the photoperiod was set as 12:12 light:dark. After 5 to 7 days, anthracnose symptoms developed on the surface of the inoculated fruit, whereas the control fruits remained healthy. The diseased fruits exhibited brown necrotic lesions, whereas the upper surfaces of the control fruits remained asymptomatic. The morphological and molecular characteristics of the reisolated pathogen were identical to those of the original fungus isolated from the natural fruit. C. siamense has been reported to cause anthracnose in the southeastern United States (Xavier et al. 2019). The pathogen causing anthracnose on pomegranates has been reported to be Colletotrichum fructicola in China (Hu et al. 2023). To our knowledge, this is the first report of anthracnose on pomegranate fruits caused by C. siamense in China. This disease can directly affect the quality and yield of the fruit. Thus, information about the characteristics of this disease could provide a theoretical basis for its prevention and control.