Unraveling the effect of phenolic extract derived from olive mill solid wastes on agro-physiological and biochemical traits of pomegranate and its associated rhizospheric soil properties.

Agricultural waste management poses a significant challenge in circular economy strategies. Olive mill wastes (OMW) contain valuable biomolecules, especially phenolic compounds, with significant agricultural potential. Our study evaluate the effects of phenolic extract (PE) derived from olive mill solid wastes (OMSW) on pomegranate agro-physiological and biochemical responses, as well as soil-related attributes. Pomegranate plants were treated with PE at doses of 100 ppm and 200 ppm via foliar spray (L100 and L200) and soil application (S100 and S200). Results showed increased biomass with PE treatments, especially with soil application (S100 and S200). Proline and soluble sugar accumulation in leaves suggested plant adaptation to PE with low-level stress. Additionally, PE application reduced malondialdehyde (MDA) and hydrogen peroxide (H2O2) contents. Higher doses of PE (S200) significantly improved net photosynthesis (Pn), transpiration rate (E), water use efficiency (WUEi), and photosynthetic efficiency (fv/fm and PIabs). Furthermore, PE treatments enhanced levels of chlorophylls, carotenoids, polyphenols, flavonoids, and antioxidant activity. Soil application of PE also increased soil enzyme activities and microbial population. Our findings suggest the beneficial impact of PE application on pomegranate agro-physiological responses, laying the groundwork for further research across various plant species and soil types to introduce nutrient-enriched PE as an eco-friendly biostimulant.

Chitosan/Pomegranate Seed Oil Emulgel Composition as a New Strategy for Dermal Delivery of Hydrocortisone.

Multifunctional delivery systems capable of modulating drug release and exerting adjunctive pharmacological activity have attracted particular attention. Chitosan (CS) and pomegranate seed oil (PO) appear to be attractive bioactive components framing the strategy of complex therapy and multifunctional drug carriers. This research is aimed at evaluating the potential of CS in combination with PO in studies on topical emulgels containing hydrocortisone as a model anti-inflammatory agent. Its particular goal was to distinguish alterations in anti-inflammatory action followed with drug dissolution or penetrative behavior between the designed formulations that differ in CS/PO weight ratio. All formulations favored hydrocortisone release with up to a two-fold increase in the drug dissolution rate within first 5 h as compared to conventional topical preparations. The clear effect of CS/PO on the emulgel biological performance was observed, and CS was found to be prerequisite for the modulation of hydrocortisone absorption and accumulation. In turn, a greater amount of PO played the predominant role in the inhibition of hyaluronidase activity and enhanced the anti-inflammatory effect of preparation E-3. Emulgels showed a negligible reduction in mouse fibroblasts' L929 cell viability, confirming their non-irritancy with skin cells. Overall, the designed formulation with a CS/PO ratio of 6:4 appeared to be the most promising topical carrier for the effective treatment of inflammatory skin diseases among the tested subjects.

CHARACTERISATION OF RESORBABLE AND NON- RESORBABLE SUTURES COATED WITH Punica granatum SEED EXTRACT: AN in vitro PILOT STUDY.

BACKGROUND: In order to achieve uneventful and rapid healing of the tissues, a suture material should be biocompatible, easy to handle, sterile, and have good and uniform tensile strength. Hence, in the present study, characterization of the suture materials was done through a novel green chemistry approach using Punica granatum seed extract. MATERIALS AND METHODS: Ethanolic extract of P. granatum seed was prepared by dissolving 25 g of P. granatum seed powder with 100 mL of ethanol. The obtained extract was coated in silk and Vicryl suture material and was tested for its surface morphology (SEM), tensile strength, anti-microbial activity, biocompatibility, and wound healing potential. RESULTS: Silk and Vicryl sutures coated with P. granatum seed extract showed the uniform coating and deposition of extract with sustaining integrity. Vicryl suture coated with the extract had good tensile strength and antimicrobial activity. The in vitro scratch assay and biocompatibility test showed that the P. granatum seed extract had excellent wound healing potential and can be used without any effect on the viability of the normal cells. CONCLUSION: Within the limitations of the study it can be concluded that P. granatum seed extract coated Vicryl sutures had good tensile strength and anti-microbial activity. P. granatum seed extract also showed excellent biocompatibility and wound healing potential.

Exploring the therapeutic potential of pomegranate juice for uterine relaxation.

BACKGROUND: The effects of pomegranate juice (PJ) and its components on uterine smooth muscle are unknown. Hence, this study unequivocally demonstrates that pomegranate juice (PJ) significantly impacts myometrial function, providing crucial insights into its relaxant properties and their potential therapeutic applications for uterine-related disorders. RESEARCH DESIGN AND METHODS: Rat uterine smooth muscle horn strips were suspended in Krebs solution organ baths. Contractions were measured isometrically using a transducer (AD instrument Australia). The effects of PJ were evaluated on contractile activity elicited by potassium chloride (KCl 60 Mm) depolarization. Inhibitors of nitric oxide (L-NAME 3 X 10-4), guanylate cyclase (methylene blue 1 X 10-5), and Prostaglandin I2 (indomethacin 3 X 10-5), as well as Potassium Channels blockers, were determined. RESULTS: The juice at concentrations from 1.5-5 mg/ml significantly decreased the rat uterine horn contraction induced by KCl. The NO, cGMP, and PGI2 inhibitors did not block the relaxation response. Furthermore, the PGI2 inhibitor significantly enhanced the relaxation effects; K+ channel blockers had no inhibitory effects on the relaxation responses. Contrarily, GLIB improved considerably relaxation. CONCLUSION: Research suggests pomegranate juice's active ingredient may reduce uterine contractions and treat uterotonic disorders, potentially preventing preterm birth and dysmenorrhea. Further research is needed to determine its mechanism of action. TRIAL REGISTRATION: Code: AEC-013.

The effect of pomegranate juice supplementation on C-reactive protein levels: GRADE-assessed systematic review and dose-response updated meta-analysis of data from randomized controlled trials.

Pomegranate juice (PJ) has a possible anti-inflammatory effect because of its polyphenol content and antioxidants. However, the anti-inflammatory effect of PJ in randomized controlled trials (RCTs) has not been consistent. A previous meta-analysis conducted in 2016 reported a nonsignificant lowering effect of PJ on C-reactive protein (CRP) levels. This systematic review and meta-analysis aim to update the pooled effect size of PJ supplementation on CRP levels in RCT studies. PubMed, Scopus, and Web of Science databases were comprehensively searched until July 2023. Eligible studies were found by screening, their relevant data was extracted, and a risk of bias assessment was performed. The pooled effect size was calculated using a random effect model as the weighted mean difference (WMD) with a 95% confidence interval. This systematic review included 11 studies with 13 effect sizes and 696 participants. Meta-analysis showed that PJ supplementation led to a significant decrease in CRP levels compared to control groups (WMD: -2.55 mg/L; 95%CI: -3.44 to -1.66; p < 0.001). Subgroup analysis demonstrated the significant reduction effect of PJ on CRP levels in studies conducted on the both sexes or only females as well as Iranian population, individuals with 40 years≤, type 2 diabetes, polycystic ovary syndrome, or trials that intervened with PJ dosage of <250 ml/day. Meta-regression and dose-response analysis reported a nonsignificant linear and nonlinear relationship between intervention characteristics (duration and dose of PJ) and CRP changes. The current meta-analysis revealed that PJ supplemantation has a beneficial effect in improving CRP levels. It is recommended to understand this effect better, and find the optimal dose and duration of PJ supplementation to reduce CRP levels in the blood, and repeat meta-analysis after related RCTs are available. For the final proof of these effects, more detailed human studies are needed.

The Effects of Pomegranate Seed Oil on Mild Cognitive Impairment.

Background: In recent years, there has been a growing interest, supported by many experimental and clinical studies, about the benefits of pomegranate in preventing various pathologic conditions, including brain neurodegeneration. The pomegranate seed oil (PSO) contains high levels of fatty acids that have antioxidant and anti-inflammatory properties. Objective: Due to the lack of clinical trials, the aim of the present study was to investigate the effects of PSO on cognition of people with mild cognitive impairment (MCI). Methods: Eighty people with the diagnosis of MCI were randomized forty to take 5 drops of PSO and follow the Mediterranean Diet (MeDi) and forty just followed MeDi. All were examined with an extensive neuropsychological assessment before and after one year of treatment. Results: The results showed that the participants who took the PSO had statistically significantly better global cognition (p = 0.004), verbal episodic memory (p = 0.009), and processing and executive functions (p < 0.001) in contrast with the participants who did not take it. Conclusions: In conclusion, the PSO can be beneficial for people with MCI as it is helpful for some important cognitive domains. As PSO is a natural product that does not burden the human body, it can be used by people with MCI and be a significant and promising part of holistic approaches for the prevention of dementia.

Ultra-High-Performance Liquid Chromatography-Electrospray Ionization-High-Resolution Mass Spectrometry for Distinguishing the Origin of Ellagic Acid Extracts: Pomegranate Peels or Gallnuts.

Ellagic acid, known for its various biological activities, is widely used. Ellagic acid from pomegranate peels is safe for consumption, while that from gallnuts is only suitable for external use. However, there is currently no effective method to confirm the source of ellagic acid. Therefore, this study establishes an analysis method using ultra-high-performance liquid chromatography-electrospray ionization-high-resolution mass spectrometry (UHPLC-ESI-HR-MS) to identify the components of crude ellagic acid extracts from pomegranate peels and gallnuts. The analysis revealed that there was a mix of components in the crude extracts, such as ellagic acid, palmitic acid, oleic acid, stearic acid, and 9(10)-EpODE. Furthermore, it could be observed that ellagic acid extracted from gallnuts contained toxic substances such as anacardic acid and ginkgolic acid (15:1). These components could be used to effectively distinguish the origin of ellagic acid from pomegranate peels or gallnuts. Additionally, a rapid quantitative analysis method using UHPLC-ESI-MS with multiple reaction monitoring (MRM) mode was developed for the quality control of ellagic acid products, by quantifying anacardic acid and ginkgolic acid (15:1). It was found that one of three ellagic acid health care products contained ginkgolic acid (C15:1) and anacardic acid at more than 1 ppm.

Pomegranate extract-loaded sphingosomes for the treatment of cancer: Phytochemical investigations, formulation, and antitumor activity evaluation.

AIM: Formulation of Pomegranate Extracts (PE)-loaded sphingosomes as an antitumor therapy for the intravenous and passive targeted delivery to various tumor types, especially that of the breast, colon, and uterus; to increase the therapeutic activity and decrease the adverse effects profile. METHODS: The pericarp and seeds' juice of Punica granatum were each extracted using D.W. and ethanol. Phytochemical investigation of all extracts was carried out including total phenolics, flavonoids, and anthocyanins contents, the antioxidant activity, as well as HPLC analysis of phenolics and flavonoids. The antitumor potential of all extracts was also tested utilizing three cell lines: MCF-7, HeLa, and HCT116. The candidate extract was chosen for the formulation phase and was entrapped into the sphingosomes using the thin-film hydration method and employing three different PE: lipids weight ratios. The synthesized formulations were characterized for their size, morphological features, zeta potential, entrapment efficiency, and in vitro drug release and kinetics modeling studies. The optimized formula was further analyzed by FTIR spectroscopy and electron microscopy. The antitumor activity of F2 was also investigated using the same cancer cell lines compared to the plant extract. RESULTS: The highest phenolics, flavonoids, and anthocyanins contents were observed in the ethanolic pericarps extract (EPE), followed by the ethanolic seeds extract (ESE). Consequently, EPE showed a higher antitumor activity hence it was selected for the formulation phase. PE-loaded sphingosomes formula (F2) was selected for having the highest EE% (71.64%), and a sustained release profile with the highest in vitro release (42.5±9.44%). By employing the DDSolver, the Weibull model was found the most suitable to describe the PE release kinetics compared to other models. The release mechanism was found to follow Fickian diffusion. Simulated pharmacokinetic parameters have portrayed F2 as the candidate formula, with the highest AUC (536.095) and slowest MDT (0.642 h). In addition, F2 exhibited a significant (p>0.05) stronger and prolonged anticancer effect against MCF-7, HeLa, and HCT116 cell lines at all concentrations tested compared to the free extract. CONCLUSION: The results proved that sphingosomes are an effective delivery system, improving pharmacological efficacy and reducing serious side effects of anticancer medications and natural products.

Traditional herbal pair Portulacae Herba and Granati Pericarpium alleviates DSS-induced colitis in mice through IL-6/STAT3/SOCS3 pathway.

BACKGROUND: Portulacae Herba and Granati Pericarpium pair (PGP) is a traditional Chinese herbal medicine treatment for colitis, clinically demonstrating a relatively favorable effect on relieving diarrhea and abnormal stools. However, the underlying mechanism remain uncertain. PURPOSE: The present study intends to evaluate the efficacy of PGP in treating colitis in mice and investigate its underlying mechanism. METHODS: The protective effect of PGP against colitis was determined by monitoring body weight, colon length, colon weight, and survival rate in mice. Colonic inflammation was assessed by serum cytokine levels, colonic H&E staining, and local neutrophil infiltration. The reversal of intestinal epithelial barrier damage by PGP was subsequently analyzed with Western blot and histological staining. Furthermore, RNA-seq analysis and molecular docking were performed to identify potential pathways recruited by PGP. Following the hints of the transcriptomic results, the role of PGP through the IL-6/STAT3/SOCS3 pathway in DSS-induced colitis mice was verified by Western blot. RESULTS: DSS-induced colitis in mice was significantly curbed by PGP treatment. PGP treatment significantly mitigated DSS-induced colitis in mice, as evidenced by improvements in body weight, DAI severity, survival rate, and inflammatory cytokines levels in serum and colon. Moreover, PGP treatment up-regulated the level of Slc26a3, thereby increasing the expressions of the tight junction/adherens junction proteins ZO-1, occludin and E-cadherin in the colon. RNA-seq analysis revealed that PGP inhibits the IL-6/STAT3/SOCS3 pathway at the transcriptional level. Molecular docking indicated that the major components of PGP could bind tightly to the proteins of IL-6 and SOCS3. Meanwhile, the result of Western blot revealed that the IL-6/STAT3/SOCS3 pathway was inhibited at the protein level after PGP administration. CONCLUSION: PGP could alleviate colonic inflammation and reverse damage to the intestinal epithelial barrier in DSS-induced colitis mice. The underlying mechanism involves the inhibition of the IL-6/STAT3/SOCS3 pathway.

Whey protein isolate and inulin-glycosylated conjugate affect the physicochemical properties and oxidative stability of pomegranate seed oil emulsion.

Glycosylated protein was obtained by the reaction of whey protein isolate(WPI) with inulin of different polymerization degrees and was used to stabilize a pomegranate seed oil emulsion. The physicochemical and antioxidative properties of the emulsions were assessed, and the impacts of accelerated oxidation on pomegranate seed oil were examined. The interfacial tension of WPI and short-chain inulin (SCI)-glycosylated conjugate (WPI-SCI) gradually decreased with increasing glycosylation reaction time. Emulsions stabilized by WPI-SCI (72 h) were the most stable, with a thick interfacial film on the surface of the droplets. After accelerated oxidation for 72 h, WPI-SCI inhibited the oxidation of oil in the emulsion. GC-IMS results showed that the production of harmful volatile components in oil was inhibited, and the peroxide strength was less than 30 mmol/kg oil. This study contributes to understanding of stable storage of lipids.

Improved Cardiovascular Effects of a Novel Pomegranate Byproduct Extract Obtained through Hydrodynamic Cavitation.

The healthy properties of pomegranate fruit, a highly consumed food, have been known for a long time. However, the pomegranate supply chain is still rather inefficient, with the non-edible fraction, whose weight is roughly half the total and is endowed with plenty of valuable bioactive compounds, either disposed of or underutilized. A novel extract obtained from non-edible byproducts (called PPE), using hydrodynamic cavitation, a green, efficient, and scalable technique, was investigated for its cardiovascular effects in vivo. PPE showed efficacy in an acute phenylephrine (PE)-induced hypertensive rat model, similar to the extract of whole fruit (PFE) obtained using the same extractive technique, along with good intestinal bioaccessibility after oral administration. Finally, when chronically administered for 6 weeks to spontaneously hypertensive rats, PPE was shown to significantly contain the increase in systolic blood pressure, comparable to the reference drug Captopril, and at a dose remarkably lower than the reported effective dose of ellagic acid. The extract from the non-edible fraction of the pomegranate fruit also showed good anti-inflammation and anti-fibrotic effects. The findings of this study, along with the extraction technique, could contribute to enhancing the value of the pomegranate supply chain, relieve the related environmental burden, and potentially improve public health.

Punicalagin, a pomegranate polyphenol sensitizes the activity of antibiotics against three MDR pathogens of the Enterobacteriaceae.

BACKGROUND: Multidrug resistance (MDR) in the family Enterobacteriaceae is a perniciously increasing threat to global health security. The discovery of new antimicrobials having the reversing drug resistance potential may contribute to augment and revive the antibiotic arsenal in hand. This study aimed to explore the anti-Enterobacteriaceae capability of bioactive polyphenols from Punica granatum (P. granatum) and their co-action with antibiotics against clinical isolates of Enterobacteriaceae predominantly prevalent in South Asian countries. METHODS: The Kandhari P. granatum (Pakistani origin) extracts were tested for anti-Enterobacteriaceae activity by agar well diffusion assay against MDR Salmonella enterica serovar Typhi, serovar Typhimurium and Escherichia coli. Predominant compounds of active extract were determined by mass spectrometry and screened for bioactivity by agar well diffusion and minimum inhibitory concentration (MIC) assay. The active punicalagin was further evaluated at sub-inhibitory concentrations (SICs) for coactivity with nine conventional antimicrobials using a disc diffusion assay followed by time-kill experiments that proceeded with SICs of punicalagin and antimicrobials. RESULTS: Among all P. granatum crude extracts, pomegranate peel methanol extract showed the largest inhibition zones of 25, 22 and 19 mm, and the MICs as 3.9, 7.8 and 7.8 mg/mL for S. typhi, S. typhimurium and E. coli, respectively. Punicalagin and ellagic acid were determined as predominant compounds by mass spectrometry. In plate assay, punicalagin (10 mg/mL) was active with hazy inhibition zones of 17, 14, and 13 mm against S. typhi, S. typhimurium and E. coli, respectively. However, in broth dilution assay punicalagin showed no MIC up to 10 mg/mL. The SICs 30 µg, 100 µg, and 500 µg of punicalagin combined with antimicrobials i.e., aminoglycoside, ß-lactam, and fluoroquinolone act in synergy against MDR strains with % increase in inhibition zone values varying from 3.4 ± 2.7% to 73.8 ± 8.4%. In time-kill curves, a significant decrease in cell density was observed with the SICs of antimicrobials/punicalagin (0.03-60 µg/mL/30, 100, 500 µg/mL of punicalagin) combinations. CONCLUSIONS: The P. granatum peel methanol extract exhibited antimicrobial activity against MDR Enterobacteriaceae pathogens. Punicalagin, the bacteriostatic flavonoid act as a concentration-dependent sensitizing agent for antimicrobials against Enterobacteriaceae. Our findings for the therapeutic punicalagin-antimicrobial combination prompt further evaluation of punicalagin as a potent activator for drugs, which otherwise remain less or inactive against MDR strains.

The effects of pomegranate consumption on blood pressure in adults: A systematic review and meta-analysis.

Considering the main component of cardiovascular disease and due to the high prevalence of hypertension, controlling blood pressure is required in individuals with various health conditions. Randomized clinical trials (RCTs) which studied the effects of pomegranate consumption on blood pressure have shown inconsistent findings. As a result, we intended to assess the effects of pomegranate consumption on systolic (SBP) and diastolic (DBP) blood pressure in adults. Systematic literature searches up to January 2024 were carried out using electronic databases, including PubMed, Web of Science, and Scopus, to identify eligible RCTs assessing the effects of pomegranate on blood pressure as an outcome. All the individuals who took part in our research were adults who consumed pomegranate in different forms as part of the study intervention. Heterogeneity tests of the selected trials were performed using the I2 statistic. Random effects models were assessed based on the heterogeneity tests, and pooled data were determined as the weighted mean difference (WMD) with a 95% confidence interval (CI). Of 2315 records, 22 eligible RCTs were included in the current study. Our meta-analysis of the pooled findings showed that pomegranate consumption significantly reduced SBP (WMD: -7.87 mmHg; 95% CI: -10.34 to -5.39; p < 0.001) and DBP (WMD: -3.23 mmHg; 95% CI: -5.37 to -1.09; p = 0.003). Individuals with baseline SBP > 130 mmHg had a significantly greater reduction in SBP compared to individuals with baseline SBP < 130 mmHg. Also, there was a high level of heterogeneity among studies (SBP: I2 = 90.0% and DBP: I2 = 91.8%). Overall, the results demonstrated that pomegranate consumption lowered SBP and DBP in adults. Although our results suggest that pomegranate juice may be effective in reducing blood pressure in the pooled data, further high-quality studies are needed to demonstrate the clinical efficacy of pomegranate consumption.

Hepatoprotective effect of methanol fruit extract of Punica granatum L in highly active antiretroviral therapy-induced toxicity in Wistar rats.

Prolonged use of Highly Active Antiretroviral Therapy (HAART) has been linked to toxicity, particularly hepatotoxicity. There are few effective drugs for HAART patients that promote hepatic cell regeneration and prevent liver injury. Therefore, the purpose of this study was to investigate the hepato-protective activity of Methanol fruit extract of Punica granatum (MFEPG) in HAART-administered rats. Thirty rats weighing between 150-200 g were randomly divided into six groups and each group comprised of five rats. Distilled water was given to the rats in group one. Only HAART was given to the rats in group two. MFEPG at doses of 100 and 400 mg/kg was given to the rats in groups three and four. MFEPG dosages of 100 and 400 mg/kg along with HAART were given to the rats in groups five and six, respectively. All treatments were via oral gavage daily for 40 days. Under halothane anesthesia, all rats were sacrificed on day 41. Liver tissues were utilized for lipid peroxidation marker; Malondialdehyde (MDA), antioxidant enzymes; Superoxide dismutase (SOD) and Catalase (CAT) and histological evaluation, while blood samples were examined for biochemical parameters (AST, ALT, ALP, Total cholesterol, Total protein, and Albumin). The HAART-treated group exhibited a significantly higher amount of the lipid peroxidation end product; MDA, and significantly lower levels of antioxidant enzymes; SOD, and CAT. Liver enzymes and total cholesterol were significantly increased with a significant reduction in Total protein and Albumin levels in the HAART-treated group. Conversely, the liver function biomarkers were returned to normal levels in the HAART and MFEPG-treated groups. Histopathological studies revealed that when HAART-exposed rats were treated with MFEPG, both the biochemical and histological results significantly improved. Thus, the antioxidant activity of MFEPG provides protection against HAART-induced liver oxidative damage. More research is needed to determine the safety of using MFEPG in humans.

The potential of postharvest zinc treatment for preservation of pomegranate aril quality.

A short shelf life usually limits the distribution and supply of pomegranate arils. Since zinc (Zn) has an indispensable role in the nutrient integrity of our diet and is effective in suppressing pathogens, this study was done as two separate experiments of pre-harvest spraying and postharvest dipping of arils with two zinc supplements, including nano zinc oxide (nZnO) and zinc sulfate (ZnSO4). The optimized concentration of both sources was used in the experiment. The pre-harvest treatment failed to extend the shelf life of arils, and, ultimately, the arils decayed after 15 days. However, the postharvest zinc treatment significantly (P < 0.01) affected all measured indices. Also, zinc sulfate was more effective than nZnO. Zn uptake was higher in postharvest treatments because exogenous Zn was in direct contact with the aril surface. After dissolving in water, Zn ions in sulfate bind to the membrane of microorganisms and thus delay cell division and microbial growth cycle. The solubility of zinc oxide nanoparticles in water is poor. Using the ZnSO4 treatment (0.8%W/V) effectively maintained the values of titratable acidity (TA), total phenolic content (TPC), total soluble solids (TSS), anthocyanin content, and antioxidant activity. Also, this treatment significantly controlled weight loss in the arils.

Development of novel carboxymethyl cellulose/gelatin-based edible films with pomegranate peel extract as antibacterial/antioxidant agents for beef preservation.

Novel antioxidant and antibacterial composite films were fabricated by incorporating pomegranate peel extract (PPE) into gelatin and carboxymethyl cellulose matrices. Increasing PPE concentration significantly (p < 0.05) altered physical properties and improved UV (decrease in light transmission 87.30 % to 9.89 % at 400 nm) and water resistance, while FTIR and molecular docking results revealed hydrogen bonding between PPE and film matrix. PPE incorporation enhanced antioxidant activity up to 84.15 ± 0.12 % and also restricted gram-positive and gram-negative bacterial growth by 72.4 % and 65.9 % respectively after 24 h, measured by antimicrobial absorption assays. For beef packaging applications at refrigeration temperatures, PPE films were most effective at extending shelf-life up to 3 days, as evidenced by reduced total viable counts, total volatile basic nitrogen, weight loss, and pH changes compared to control films. Therefore, these antioxidant and antibacterial films have potential applications in food packaging to protect against mechanical stress, light exposure, microbial spoilage, and oxidative free radicals.

Pomegranate extract in polyphosphate-fluoride mouthwash reduces enamel demineralization.

OBJECTIVES: To evaluate the anti-demineralizing effect of a mouthwash comprising pomegranate peel extract (PPE 3%), sodium trimetaphosphate (TMP 0.3%), and fluoride (F 225 ppm) in an in situ study, and to assess its irritation potential in an ex vivo study. METHODS: This double-blind crossover study was conducted in four phases with 7 days each. Twelve volunteers used palatal appliances containing enamel blocks, which were subjected to cariogenic challenges. The ETF formulation (PPE + TMP + F, pH 7.0), TF formulation (TMP + F, pH 7.0), deionized water (W, pH 7.0), and essential oil commercial mouthwash (CM, 220 ppm F, pH 4.3) were dropped onto the enamel twice daily. The percentage of surface hardness loss, integrated loss of subsurface hardness, calcium, phosphorus, and fluoride in enamel and biofilms were determined. In addition, alkali-soluble extracellular polysaccharide concentrations were analyzed in the biofilms. The irritation potential was evaluated using the hen's egg chorioallantoic membrane test through the vascular effect produced during 300-s of exposure. RESULTS: ETF was the most efficacious in preventing demineralization. It also showed the highest concentrations of calcium and phosphorus in the enamel and in the biofilm, as well as the lowest amount of extracellular polysaccharides in the biofilm. In the eggs, ETF produced light reddening, whereas CM led to hyperemia and hemorrhage. CONCLUSIONS: The addition of PPE to formulations containing TMP and F increased its anti-demineralizing property, and this formulation presented a lower irritation potential than the CM. CLINICAL RELEVANCE: ETF can be a promising alternative alcohol-free mouthwash in patients at high risk of caries.

Engineering Yarrowia lipolytica for Sustainable Production of the Pomegranate Seed Oil-Derived Punicic Acid.

Punicic acid is a conjugated linolenic acid with various biological activities including antiobesity, antioxidant, anticancer, and anti-inflammatory effects. It is often used as a nutraceutical, dietary additive, and animal feed. Currently, punicic acid is primarily extracted from pomegranate seed oil, but it is restricted due to the extended growth cycle, climatic limitations, and low recovery level. There have also been reports on the chemical synthesis of punicic acid, but it resulted in a mixture of structurally similar isomers, requiring additional purification/separation steps. In this study, a comprehensive strategy for the production of punicic acid in Yarrowia lipolytica was implemented by pushing the supply of linoleic acid precursors in a high-oleic oil strain, expressing multiple copies of the fatty acid conjugase gene from Punica granatum, engineering the acyl-editing pathway to improve the phosphatidylcholine pool, and promoting the assembly of punicic acid in the form of triglycerides. The optimal strain with high oil production capacity and a significantly increased punicic acid ratio accumulated 3072.72 mg/L punicic acid, accounting for 6.19% of total fatty acids in fed-batch fermentation, providing a viable, sustainable, and green approach for punicic acid production to substitute plant extraction and chemical synthesis production.

Use of encapsulated pomegranate seed oil in novel coarse and nanosized materials for improving the storage life of strawberry.

Different-sized pomegranate seed oil-based emulsions (coarse (CsP) and nanoemulsions (NsP): 1246 and 325 nm) were successfully prepared. Strawberries treated with NsP and CsP showed a significant decrease (p < 0.05) in yeast-mold counts (TMY) by 1.80 log CFU g-1, and mesophilic aerobic bacteria counts (TMAB) decreased (p < 0.05) by 0.91 log CFU g-1, respectively. CsP- and NsP-treated strawberries had a TPC of 74.45 and 82.35 mg GAE kg-1, respectively, while control samples had a TPC of 44.24 mg GAE kg-1. The strawberries treated with NsP exhibited the highest antioxidant capacity with 179.44 mol TEAC g-1. After treatment with a coarse emulsion, severity levels of A. niger and B. cinerea were 60 and 73 % while the nanoemulsion treatment significantly reduced severity levels to 55.3 and 56 %. The coarse and nanoemulsions may have potential use within the food industry owing to their antioxidant and antifungal properties as well as their ability to enhance strawberry quality and function.

Phytochemical Extract from Carica papaya Leaves and Punica granatum Seeds as Therapy Against Cognitive Impairment in a Murine Model.

Mild cognitive impairment (MCI) is defined as inter-stage between normal cognitive aging and major neurocognitive disorder (MND). This state of decay is a crucial factor in treatment to prevent the progression to MND. In this study, our group developed a virtual screening process to evaluate 2568 phytochemical compounds against 5 key proteins associated with MCI and MND. As a result, two potential candidates were identified: carpaine, found in Carica papaya leaves, and punicalagin, present in Punica granatum. A model of cognitive impairment (CI) was developed in 10-month-old male Sprague Dawley rats by administering aluminum chloride (AlCl3) at a dose of 100 mg/kg/day for 30 days. After AlCl3 administration period, one of the groups received carpaine and punicalagin in a phytochemical extract (PE) by oral gavage for 30 days. Novel object recognition test (NOR) was assessed at three different time points (T1 - before CI, T2 - after CI, and T3 - after PE treatment). Glial fibrillary acidic protein (GFAP) and neurofilament light chain (NfL) were identified in the hippocampus of rats at the end of the study period. After administration of AlCl3, a reduction in discrimination index vs control rats (CI = 0.012 ± 0.08 vs Control = 0.076 ± 0.03), was observed. After phytochemical extract treatment, a significant increase in discrimination index values was observed in the PE group 0.4643 ± 0.13 vs CI group 0.012 ± 0.08. Additionally, the evaluation of immunohistochemistry showed an increase in GFAP positivity in the hippocampus of the CI groups, while a slight decrease was observed in the PE group. This work addressed a comprehensive methodology that utilized in silico tools to identify phytochemical compounds (carpaine and punicalagin) as potential candidates for affecting key proteins in CI. The phytochemical extract containing carpaine and punicalagin resulted in a trend in the decrease of GFAP expression in the hippocampus and improved recognition memory in rats with CI induced by age and AlCl3 administration.