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The challenges in the production of metabolites of medicinal potential from wild plants include low yields, slow growth rates, seasonal variations, genetic variability and regulatory as well as ethical constraints. Overcoming these challenges is of paramount significance and interdisciplinary approaches and innovative strategies are prevalently applied to optimize phytoconstituents' production, enhance yield, biomass, ensure sustainable consistency and scalability. In this study, we investigated the effects of elicitation with yeast extract and calcium oxide nanoparticles (CaONPs) on in vitro cultures of Swertia chirata (Roxb. ex Fleming) Karsten. Specifically, we examined the effects of different concentrations of CaONPs in combination with different concentrations of yeast extract on various parameters related to callus growth, antioxidant activity, biomass and phytochemical contents. Our results showed that elicitation with yeast extract and CaONPs had significant effects on the growth and characteristics of callus cultures of S. chirata . The treatments involving yeast extract and CaONPs were found to be the most effective in increasing the contents of total flavonoid contents (TFC), total phenolic contents (TPC), amarogentin and mangiferin. These treatments also led to an improvement in the contents of total anthocyanin and alpha tocopherols. Additionally, the DPPH scavenging activity was significantly increased in the treated samples. Furthermore, the treatments involving elicitation with yeast extract and CaONPs also led to significant improvements in callus growth and characteristics. These treatments promoted callus response from an average to an excellent level and improved the color and nature of the callus from yellow to yellow-brown and greenish and from fragile to compact, respectively. The best response was observed in treatments involving 0.20 g/L yeast extract and 90 ug/L CaONPs. Overall, our findings suggest that elicitation with yeast extract and CaONPs can be a useful strategy for promoting the growth, biomass, phytochemical contents and antioxidant activity of callus cultures of S. chirata in comparison to wild plant herbal drug samples.

Iron oxide nanoparticles (NPs) have attracted substantial interest due to their superparamagnetic features, biocompatibility, and nontoxicity. The latest progress in the biological production of Fe 3 O 4 NPs by green methods has improved their quality and biological applications significantly. In this study, the fabrication of iron oxide NPs from Spirogyra hyalina and Ajuga bracteosa was conducted via an easy, environmentally friendly, and cost-effective process. The fabricated Fe 3 O 4 NPs were characterized using various analytical methods to study their unique properties. UV-Vis absorption peaks were observed in algal and plant-based Fe 3 O 4 NPs at 289 nm and 306 nm, respectively. Fourier transform infrared (FTIR) spectroscopy analyzed diverse bioactive phytochemicals present in algal and plant extracts that functioned as stabilizing and capping agents in the fabrication of algal and plant-based Fe 3 O 4 NPs. X-ray diffraction of NPs revealed the crystalline nature of both biofabricated Fe 3 O 4 NPs and their small size. Scanning electron microscopy (SEM) revealed that algae and plant-based Fe 3 O 4 NPs are spherical and rod-shaped, averaging 52 nm and 75 nm in size. Energy dispersive X-ray spectroscopy showed that the green-synthesized Fe 3 O 4 NPs require a high mass percentage of iron and oxygen to ensure their synthesis. The fabricated plant-based Fe 3 O 4 NPs exhibited stronger antioxidant properties than algal-based Fe 3 O 4 NPs. The algal-based NPs showed efficient antibacterial potential against E. coli , while the plant-based Fe 3 O 4 NPs displayed a higher zone of inhibition against S. aureus . Moreover, plant-based Fe 3 O 4 NPs exhibited superior scavenging and antibacterial potential compared to the algal-based Fe 3 O 4 NPs. This might be due to the greater number of phytochemicals in plants that surround the NPs during their green fabrication. Hence, the capping of bioactive agents over iron oxide NPs improves antibacterial applications.

Modern nanotechnology encompasses every field of life. Nowadays, phytochemically fabricated nanoparticles are being widely studied for their bioactivities and biosafety. The present research studied the synthesis, characterization, stability, biocompatibility, and in vitro bioactivities of calcium oxide nanoparticles (CaONPs). The CaONPs were synthesized using Citrullus colocynthis ethanolic fruit extracts. Greenly synthesized nanoparticles had an average size of 35.93 ± 2.54 nm and showed an absorbance peak at 325 nm. An absorbance peak in this range depicts the coating of phenolic acids, flavones, flavonols, and flavonoids on the surface of CaONPs. The XRD pattern showed sharp peaks that illustrated the preferred cubic crystalline nature of triturate. A great hindrance to the use of nanoparticles in the field of medicine is their extremely reactive nature. The FTIR analysis of the CaONPs showed a coating of phytochemicals on their surface, due to which they showed great stability. The vibrations present at 3639 cm -1 for alcohols or phenols, 2860 cm -1 for alkanes, 2487 cm -1 for alkynes, 1625 cm -1 for amines, and 1434 cm -1 for carboxylic acids and aldehydes show adsorption of phytochemicals on the surface of CaONPs. The CaONPs were highly stable over time; however, their stability was slightly disturbed by varying salinity and pH. The dialysis membrane in vitro release analysis revealed consistent nanoparticle release over a 10-h period. The bioactivities of CaONPs, C. colocynthis fruit extracts, and their synergistic solution were assessed. Synergistic solutions of both CaONPs and C. colocynthis fruit extracts showed great bioactivity and biosafety. The synergistic solution reduced cell viability by only 14.68% and caused only 16% hemolysis. The synergistic solution inhibited Micrococcus luteus slightly more effectively than streptomycin, with an activity index of 1.02. It also caused an 83.87% reduction in free radicals.

Research on the use of different parts of the Moringa oleifera plant as a nutritional and pharmaceutical resource for human and animals has increased in recent years. This study aimed to investigate the chemical composition and the TPCs and TFCs of Moringa leaves, the antimicrobial activities of Moringa successive ethanolic, aqueous, crude aqueous extracts, and green-chemically synthesized characterized Ag-NPs. The results indicated that the ethanolic extract recorded the highest activity against E. coli . On the other side, the aqueous extract showed higher activity, and its effects ranged from 0.03 to 0.33 mg/mL against different strains. The MIC values of Moringa Ag-NPs against different pathogenic bacteria ranged from 0.05 mg/mL to 0.13 mg/mL, and the activity of the crude aqueous extract ranged from 0.15 to 0.83 mg/mL. For the antifungal activity, the ethanolic extract recorded the highest activity at 0.04 mg/mL, and the lowest activity was recorded at 0.42 mg/mL. However, the aqueous extract showed effects ranging from 0.42 to 1.17 mg/mL. Moringa Ag-NPs showed higher activity against the different fungal strains than the crude aqueous extract, and they ranged from 0.25 to 0.83 mg/mL. The MIC values of the Moringa crude aqueous extract ranged from 0.74 to 3.33 mg/mL. Moringa Ag-NPs and their crude aqueous extract may be utilized to boost antimicrobial attributes.

Wild edible food plants (WFPs) are valuable resources in the traditional food systems of many local cultures worldwide, particularly in underdeveloped regions. Understanding patterns of food preferences requires conducting cross-cultural food studies among various ethnic groups in a specific area. In this context, the current study aimed to record WFP use among five ethnic groups in Punjab, Pakistan, by interviewing 175 informants selected through snowball sampling. The indicator food species for different ethnic groups were calculated using indicator analysis based on the percentage of citations. A total of 71 wild food plants (WFPs) belonging to 57 genera and 27 families were observed in the study area. A high proportion of these wild food plants (WFPs) belonged to Fabaceae with eleven species (15%), followed by Moraceae with seven species (9%). Fruits were most widely used (43%), followed by leaves (19%), and shoots (16%). The majority (35 species, 49%) of plants of WFPs were eaten as cooked vegetables. A cross-cultural comparison revealed that four species overlapped among five ethnic groups (Arain, Jutt, Rajpot, Mewati, and Dogar). The Arain ethnic group gathered and consumed a remarkable number of wild plants (35 species), possibly due to a special connection with the general abundance of the local flora, and being close to nature by adopting professions more allied to WFPs in the study area. The analysis of indicator species revealed distinct significant indicator values ( p ≤ 0.05) between the main food species among the various ethnic groups. Amaranthus viridis was a common indicator of food in all five ethnic groups, while Ziziphus nammularia was a common indicator food plant of the Mewati, Rajpot, and Jutt ethnic groups; these plants are important in local diets, especially during times of food scarcity brought on by disease or drought. In addition, the current study reports 20 WFPs that have been rarely documented as human food in Pakistan's ethnobotanical literature. Future development plans should consider biocultural heritage and pay appropriate attention to local ecological knowledge, dynamics, and historical exchanges of traditional food systems.

Chronic obstructive pulmonary disease (COPD) is a major cause of death and morbidity; it may be accompanied by oxidative stress and inflammation with or without underlying genetic etiology. Finding circulating biomarkers for COPD that can help early diagnosis and predict exacerbation and association with respiratory functions has been challenging. There were 40 healthy participants and 60 COPD patients in this research. The rs2070600 gene variant was examined by PCR-RFLP. Circulating sRAGE and annexin A1 levels were determined by ELISA. GSH and MDA were determined by spectrophotometry. In COPD patients, sRAGE serum levels were substantially lower, but conversely, annexin A1 levels were much greater than in controls. The rs2070600 gene polymorphism's strong association with COPD was demonstrated by genotyping and allelic frequency distribution. The GA genotype was most distributed in COPD, and it was strongly linked to lower serum sRAGE levels. The interrelation between annexin A1, sRAGE, and COPD could be explained through effects on inflammatory mediators' pathways. The rs2070600 gene polymorphism was found to significantly enhance the risk of COPD. Serum sRAGE and annexin A1 may be considered potential diagnostic tools for COPD. Through impacts on GSH and MDA levels that alter the release of inflammatory factors and, therefore, lung damage, it is possible to explain the relationship between annexin A1, sRAGE, and COPD., Competing Interests: The authors report no competing interest.

Hyperlipidemia is still the leading cause of heart disease in patients with hypertension. The purpose of this study is to make rosuvastatin calcium (ROS) and atenolol (AT) bilayer tablets to treat coexisting dyslipidemia and hypertension with a single product. ROS was chosen for the immediate-release layer of the constructed tablets, whereas AT was chosen for the sustained-release layer. The solid dispersion of ROS with sorbitol (1:3 w / w ) was utilized in the immediate-release layer while hydroxypropyl methylcellulose (HPMC), ethylcellulose (EC), and sodium bicarbonate were incorporated into the floating sustained-release layer. The concentrations of HPMC and EC were optimized by employing 3 2 full factorial designs to sustain AT release. The bilayer tablets were prepared by the direct compression method. The immediate-release layer revealed that 92.34 ± 2.27% of ROS was released within 60 min at a pH of 1.2. The second sustained-release layer of the bilayer tablets exhibited delayed release of AT (96.65 ± 3.36% within 12 h) under the same conditions. The release of ROS and AT from the prepared tablets was found to obey the non-Fickian diffusion and mixed models (zero-order, Higuchi and Korsmeyer-Peppas), respectively. Preclinical studies using rabbit models investigated the impact of ROS/AT tablets on lipid profiles and blood pressure. A high-fat diet was used to induce obesity in rabbits. Bilayer ROS/AT tablets had a remarkable effect on decreasing the lipid profiles, slowing weight gain, and lowering blood pressure to normal levels when compared to the control group.

At present, the majority of APIs synthesized today remain challenging tasks for formulation development. Many technologies are being utilized or explored for enhancing solubility, such as chemical modification, novel drug delivery systems (microemulsions, nanoparticles, liposomes, etc.), salt formation, and many more. One promising avenue attaining attention presently is supersaturated drug delivery systems. When exposed to gastrointestinal fluids, drug concentration exceeds equilibrium solubility and a supersaturation state is maintained long enough to be absorbed, enhancing bioavailability. In this review, the latest developments in supersaturated drug delivery systems are addressed in depth.

Dietary food components have the ability to affect immune function; following absorption, specifically orally ingested dietary food containing lectins can systemically modulate the immune cells and affect the response to self- and co-administered food antigens. The mannose-binding lectins from garlic ( Allium sativum agglutinins; ASAs) were identified as immunodulatory proteins in vitro. The objective of the present study was to assess the immunogenicity and adjuvanticity of garlic agglutinins and to evaluate whether they have adjuvant properties in vivo for a weak antigen ovalbumin (OVA). Garlic lectins (ASA I and ASA II) were administered by intranasal (50 days duration) and intradermal (14 days duration) routes, and the anti-lectin and anti-OVA immune (IgG) responses in the control and test groups of the BALB/c mice were assessed for humoral immunogenicity. Lectins, co-administered with OVA, were examined for lectin-induced anti-OVA IgG response to assess their adjuvant properties. The splenic and thymic indices were evaluated as a measure of immunomodulatory functions. Intradermal administration of ASA I and ASA II had showed a four-fold and two-fold increase in anti-lectin IgG response, respectively, vs. the control on day 14. In the intranasal route, the increases were 3-fold and 2.4-fold for ASA I and ASA II, respectively, on day 50. No decrease in the body weights of animals was noticed; the increases in the spleen and thymus weights, as well as their indices, were significant in the lectin groups. In the adjuvanticity study by intranasal administration, ASA I co-administered with ovalbumin (OVA) induced a remarkable increase in anti-OVA IgG response (~six-fold; p < 0.001) compared to the control, and ASA II induced a four-fold increase vs. the control on day 50. The results indicated that ASA was a potent immunogen which induced mucosal immunogenicity to the antigens that were administered intranasally in BALB/c mice. The observations made of the in vivo study indicate that ASA I has the potential use as an oral and mucosal adjuvant to deliver candidate weak antigens. Further clinical studies in humans are required to confirm its applicability.

Before entering the cell, the SARS-CoV-2 spike glycoprotein receptor-binding domain (RBD) binds to the human angiotensin-converting enzyme 2 (hACE2) receptor. Hence, this RBD is a critical target for the development of antiviral agents. Recent studies have discovered that SARS-CoV-2 variants with mutations in the RBD have spread globally. The purpose of this in silico study was to determine the potential of a fruit bromelain-derived peptide. DYGAVNEVK. to inhibit the entry of various SARS-CoV-2 variants into human cells by targeting the hACE binding site within the RBD. Molecular docking analysis revealed that DYGAVNEVK interacts with several critical RBD binding residues responsible for the adhesion of the RBD to hACE2. Moreover, 100 ns MD simulations revealed stable interactions between DYGAVNEVK and RBD variants derived from the trajectory of root-mean-square deviation (RMSD), radius of gyration (Rg), and root-mean-square fluctuation (RMSF) analysis, as well as free binding energy calculations. Overall, our computational results indicate that DYGAVNEVK warrants further investigation as a candidate for preventing SARS-CoV-2 due to its interaction with the RBD of SARS-CoV-2 variants.

The present study aimed to analyze the phytoconstituents of Neptunia triquetra (Vahl) Benth. Anti-inflammatory and hepatoprotective activities of ethanol (EE), chloroform (CE) and dichloromethane (DCME) of stem extracts were evaluated using in vivo experimental models. The extracts were analyzed for phytoconstituents using GC-HRMS. Anti-inflammatory activity of CE, EE and DCME was accessed using carrageenan-induced paw oedema, cotton pellet-induced granuloma and the carrageenan-induced air-pouch model in Wistar albino rats. The hepatotoxicity-induced animal models were investigated for the biochemical markers in serum (AST, ALT, ALP, GGT, total lipids and total protein) and liver (total protein, total lipids, GSH and wet liver weight). In the in vivo study, animals were divided into different groups (six in each group) for accessing the anti-inflammatory and hepatoprotective activity, respectively. GC-HRMS analysis revealed the presence of 102 compounds, among which 24 were active secondary metabolites. In vivo anti-inflammatory activity of stem extracts was found in the order: indomethacin > chloroform extract (CE) > dichloromethane extract (DCME) > ethanolic extract (EE), and hepatoprotective activity of stem extracts in the order: CE > silymarin > EE > DCME. The results indicate that N. triquetra stem has a higher hepatoprotective effect than silymarin, however the anti-inflammatory response was in accordance with or lower than indomethacin.

The aim of this study is to assess the efficiency of psyllium husk ethanolic extract (PHEE) on Triton X-100 induced hyperlipidemic rats by studying the changes in hepatic and pancreatic function and histopathology. Forty male albino rats (bodyweight 175-188 g) were grouped randomly into four sets with ten rats. The experimental groups included: (1) control group (CON); (2) Triton X-100 induced hyperlipidemic group-rats were intraperitoneally injected with a single dose of Triton X-100 (100 mg/kg body weight) on the 21st day of Trial onset; (3) PHEE group-PHEE was orally administered (100 mg/kg body weight dissolved in 1 mL of distilled water) by gastric tube from the first day of the experiment until the fortieth day, once daily, (PHEE); (4) PHEE +Triton group, which received PHEE orally with the induction of hyperlipidemia. Treating hyperlipidemic rats with PHEE showed a decrease in the total serum lipids, triglyceride (TG), total cholesterol (TC), atherogenic index (AI), and malondialdehyde (MDA) with an increase in superoxide dismutase (SOD) and catalase (CAT) activities. PHEE administration alleviated the negative impact of Triton on the serum levels of glucose, insulin, glycated hemoglobin (HbA1c), homeostatic model assessment for insulin resistance (HOMA IR index), leptin hormone, Alanine Aminotransferase (ALT), Aspartate Aminotransferase (AST), Gamma-Glutamyl Transferase (GGT) and proteinogram. The Triton-induced hyperlipidemic rats showed extensive histopathological changes in the liver and pancreas, which were alleviated with PHEE administration. It could be concluded that PHEE has potent effects against hyperlipidemia, hyperglycemia, and oxidative stress due to its biologically active constituents detected by GC-MS analysis. This study's findings may help develop a novel trial against the effects of hyperlipidemia in the future.

Callus, suspension and bioreactor cultures of Verbena officinalis were established, and optimized for biomass growth and production of phenylpropanoid glycosides, phenolic acids, flavonoids and iridoids. All types of cultures were maintained on/in the Murashige and Skoog (MS) media with 1 mg/L BAP and 1 mg/L NAA. The inoculum sizes were optimized in callus and suspension cultures. Moreover, the growth of the culture in two different types of bioreactors-a balloon bioreactor (BB) and a stirred-tank bioreactor (STB) was tested. In methanolic extracts from biomass of all types of in vitro cultures the presence of the same metabolites-verbascoside, isoverbascoside, and six phenolic acids: protocatechuic, chlorogenic, vanillic, caffeic, ferulic and rosmarinic acids was confirmed and quantified by the HPLC-DAD method. In the extracts from lyophilized culture media, no metabolites were found. The main metabolites in biomass extracts were verbascoside and isoverbascoside. Their maximum amounts in g/100 g DW (dry weight) in the tested types of cultures were as follow: 7.25 and 0.61 (callus), 7.06 and 0.48 (suspension), 7.69 and 0.31 (BB), 9.18 and 0.34 (STB). The amounts of phenolic acids were many times lower, max. total content reached of 26.90, 50.72, 19.88, and 36.78 mg/100 g DW, respectively. The highest content of verbascoside and also a high content of isoverbascoside obtained in STB (stirred-tank bioreactor) were 5.3 and 7.8 times higher than in extracts from overground parts of the parent plant. In the extracts from parent plant two iridoids-verbenalin and hastatoside, were also abundant. All investigated biomass extracts and the extracts from parent plant showed the antiproliferative, antioxidant and antibacterial activities. The strongest activities were documented for the cultures maintained in STB. We propose extracts from in vitro cultured biomass of vervain, especially from STB, as a rich source of bioactive metabolites with antiproliferative, antioxidant and antibacterial properties.

Discovering new natural resources of polyphenols is the aim of many recent studies in the field of natural product research. This study tentatively investigated the polyphenols profile of the stems of seven Mammillaria species (M. rhodantha, M. spinosissima, M. hahniana, M. crucigera, M. candida, M. albilanata , and M. muehlenpfordtii ) using high performance liquid chromatography with DAD detector (HPLC-DAD) method. Furthermore, the anti-cancer, anti-oxidant, and anti-bacterial potentials of these extracts as well as major identified phenols were explored. The HPLC-DAD study confirmed the availability of six phenolic acids, including gentisic acid, chlorogenic acid, caffeic acid, protocatechuic acid, sinapic acid, and p-hydroxybenzoic acid. The dominant compounds were: gentisic acid in M. rhodantha and M. spinosissima ; chlorogenic acid in M. muehlenpfordtii, M. crucigera , and M. rhodantha; and caffeic acid in M. rhodantha, M. crucigera, and M. spinosissima. Stems of Mammillaria sp. showed antiproliferative effects against HeLa, MCF-7, and Jurkat cells. In HeLa and MCF-7 cells, the best antiproliferative activities were found in the treatments with M. rhodantha, M. spinosissima , and M. muehlenpfordtii . The apoptotic assay of M. rhodantha, M. spinosissima , and M. muehlenpfordtii showed accumulation of necrotic cells in the early and late apoptotic phase. M. rhodantha, M. spinosissima , and M. muehlenpfordtii showed the highest anti-oxidant activities using 2,2-diphenyl-1-picrylhydrazyl (DPPH), β-carotene bleaching, and ferric reducing anti-oxidant power (FRAP) assays. M. rhodantha was the best source of antioxidants. Mammillaria sp. showed moderate anti-bacterial effects against bacteria and the highest effects were found using the extracts of M. rhodantha, M. spinosissima, M. crucigera and M. muehlenpfordtii against most bacteria. The anti-bacterial activities were attributed to other phenolic compounds (e.g., chlorogenic acid) than gentisic acid, which was not active against most bacteria. Mammillaria sp. could be considered to be an important natural source of phenolic acids with anti-cancer, anti-bacterial, and anti-oxidant activities.

Tree bark represents an important source of medicinal compounds that may be useful for cancer therapy. In the current study, high-performance liquid chromatography with diode-array detection (HPLC-DAD) was used to determine the profile of the phenolic compounds of Catalpa speciosa , Taxus cuspidata , and Magnolia acuminata bark extracts. The antioxidant and anticancer bioactivities against different cancer cell lines were investigated. M. acuminata exerted significantly higher antioxidant activities in the diphenyl picrylhydrazine and β-carotene-linoleic acid assays than the other species. In C. speciosa, novel profiles of phenolic acids (ferulic acid was the predominant compound) and catechin were detected. In T. cuspidata , six phenolic acids were detected; the predominant compounds were hydroxycaffeic acid and protocatechuic acid. In M. acuminata , two phenolic acids and three catechins were detected; catechin was the predominant compound. The three species exerted clear anticancer activity against MCF-7, HeLa, Jurkat, T24, and HT-29 cells, with the strongest activity found in the extracts from M. acuminata . No antiproliferative activity against normal cells was found. Flow cytometry revealed greater accumulation of necrotic and early/late apoptotic cells in various treated cancer cells than in untreated control cells, and protocatechuic acid induced a similar accumulation of necrotic cells to that of the bark extracts. Caspase-3 and -7 activity was increased in cancer cells treated with different bark extracts; the highest activity was found in the M. acuminata treatment. Our results suggested that the treatment of cancer cells with bark extracts of M. acuminata, C. speciosa, and T. cuspidata , and protocatechuic acid induced apoptosis, suggesting an association between anticancer activities and individual phenolic compounds.

To investigate the transcriptional regulation of gene expression, chimeric fusions, between the beta-glucuronidase reporter gene (GUS) and the isolated promoter regions of the pvPDF gene (pvPDF-PRO: GUS), were constructed and introduced into Nicotiana tabacum. Analysis of transgenic pvPDF-PRO:GUS tobacco plants indicated that GUS activity was observed with all the promoter constructs with the strongest being in leaf followed by stem and roots. These results clearly demonstrate that pvPDF-PRO is a strong inducible and near-constitutive promoter and emphasize the great application potential for plant genetic engineering studies. Interestingly, a search for putative cis-acting elements in the pvPDF promoter architecture revealed the presence of some important transcription regulatory elements including: CAAT Box, TATA Box, CATA Box, and light regulatory elements (CCA1, GATA, GT-1). Taken together, these results further our understanding of the regulation of the pvPDF promoter activity.