This study explored the inhibitory effect of pomegranate flower polysaccharide (PFPS) on mastitis through in vitro and in vivo models. PFPS is a new type of polysaccharide isolated and extracted from pomegranate flowers. The result revealed that PFPS consists of GalA, Ara, and Gal, and the residues consist of 1,4-GalpA, 1,4-Galp, and 1,3,6-Galp, which contain HG-type and RG-I-type pectin structural domains. In vitro studies showed that PFPS could inhibit LPS-enhanced phagocytosis of RAW 264.7 cells and the release of IL-1β, IL-10, and TNF-α. In vivo, studies showed that PFPS improved xylene-induced mouse ear swelling and carrageenan-induced mouse paw edema by inhibiting inflammatory factors. In the mouse mastitis model, PFPS significantly improved LPS-induced inflammation and oxidative stress in mammary tissue. Intestinal flora sequencing results showed that PFPS could effectively regulate the intestinal flora of mice, reduce the relative abundance of pathogenic bacteria Oscillospira and AF12, and increase the probiotics Blautia, Parabacteroides, Allobaculum, and Clostridiaceae_Clostridium. Therefore, PFPS ultimately played a role in preventing mastitis by regulating the intestinal flora and further improving the blood-milk barrier. This study provides a scientific basis for PFPS as a potential candidate drug for the treatment of mastitis.

BACKGROUND: Punica granatum L., commonly known as pomegranate, is renowned for its health benefits, primarily associated with the consumption of its fruit and seeds. However, its non-edible parts, including leaves, have been used in traditional medicine as a remedy with anti-inflammatory and anti-diabetic properties. Considering the abundance of bioactive compounds, predominantly flavonols, flavones, and tannins P. granatum leaf (PGL) extract holds potential as health-promoting agent. Yet, its effect on longevity and healthspan remains largely unexplored.
OBJECTIVE: Our study aims to explore the potential of PGL extract to enhance healthspan and ameliorate age-related frailty in Caenorhabditis elegans. Additionally, we seek to elucidate its effect on the molecular signaling networks associated with stress resistance and longevity.
METHODS: After characterizing the extract metabolite profile by NMR spectroscopy, phenotypic and stress analyses were performed. In order to establish the molecular mechanism of action, the involvement of signaling pathways key to longevity were investigated by means of real-time quantitative PCR (RT-qPCR) and the use of transgenic strains (MIR13, MAH240, LD1, and OH16024). In addition, the effect of PGL on metabolism and lipid accumulation, as well as mitochondrial homeostasis, was examined.
RESULTS: The PGL extract supplementation significantly enhanced stress resistance and extended the lifespan of C. elegans. Additionally, it improved locomotion, as well as metabolic and mitochondrial functions, indicating an overall improvement in health. The molecular mechanisms highlight the coordinated regulation of stress response, metabolic homeostasis, and longevity signaling pathways. Specifically, our results demonstrate the essential roles of HLH-30/TFEB, in conjunction with DAF-16/FOXO and SKN-1/NRF2, as mediators of the PGL extract effect on healthspan.
CONCLUSIONS: Our findings emphasize the potential of PGL extract to ameliorate age-related decline, induce longevity and further enhance healthspan. Given the diverse effects on the molecular network associated with stress-adaptations, longevity and metabolic control, PGL extract might become a promising natural product with a particular importance to the field of gerontology.

Punica granatum L., commonly known as pomegranate, is native to Afghanistan and Iran, and today widely cultivated all over the world. Pomegranate polysaccharides are one of the most important bioactive components of P. granatum, which have a wide range of beneficial biological activities, such as anticancer, immunostimulatory, hepatoprotection, anti-psoriasis and antioxidation. Hot water extraction is currently the most commonly used method to isolate pomegranate polysaccharides. The structural characteristics of pomegranate polysaccharides have been extensively investigated through various advanced modern analytical techniques. This review focuses on the extraction, purification, structural characteristics, biological activities and structure-activity relationships of polysaccharides from Punica granatum. The aim of this article is to comprehensively and systematically summarize recent information of polysaccharides from Punica granatum and to serve as a basis for further research and development as therapeutic agents and functional foods.

The chemical and transcriptional changes in the cuticle of pomegranate (Punica granatum L.) fruit grown under different environmental conditions were studied. We collected fruit from three orchards located in different regions in Israel, each with a distinct microclimate. Fruit were collected at six phenological stages, and cutin monomers in the fruit cuticle were profiled by gas chromatography-mass spectrometry (GC-MS), along with qPCR transcript-expression analyses of selected cutin-related genes. While fruit phenotypes were comparable along development in all three orchards, principal component analyses of cutin monomer profiles suggested clear separation between cuticle samples of young green fruit to those of maturing fruit. Moreover, total cutin contents in green fruit were lower in the orchard characterized by a hot and dry climate compared to orchards with moderate temperatures. The variances detected in total cutin contents between orchards corresponded well with the expression patterns of BODYGUARD, a key biosynthetic gene operating in the cutin biosynthetic pathway. Based on our extraction protocols, we found that the cutin polyester that builds the pomegranate fruit cuticle accumulates some levels of gallic acid-the precursor of punicalagin, a well-known potent antioxidant metabolite in pomegranate fruit. The gallic acid was also one of the predominant metabolites contributing to the variability between developmental stages and orchards, and its accumulation levels were opposite to the expression patterns of the UGT73AL1 gene which glycosylates gallic acid to synthesize punicalagin. To the best of our knowledge, this is the first detailed composition of the cutin polyester that forms the pomegranate fruit cuticle.

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.

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.

Pomegranate peel extract (PPE) hydrogel films filled with citric acid (CA) and β-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 β-CD controlled the physicochemical parameters of developed PPE/CA/β-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/β-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/β-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.

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.

UNASSIGNED: 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.
UNASSIGNED: 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.
UNASSIGNED: 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.
UNASSIGNED: 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.

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.