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In the present study, we investigated the anti-biofilm effect of urinary catheters fabricated with biogenic nanoparticles synthesized from metabolites of Justicia adhatoda under in vitro conditions against human pathogenic bacteria. Silver nanoparticles were synthesized in the reaction mixture composed of 2 % w/v of 0.1 M of precursor (silver nitrate) and 0.2 g of the metabolites obtained from ethanolic extract of Justicia adhatoda. Characterization of the nanoparticles was done by UV visible spectroscopy, fourier infrared spectroscopy (FTIR), scanning electron microscopy (SEM) and X ray diffraction (XRD) to confirm the structural and functional properties. Primary conformation of nanoparticles synthesis by UV visible spectroscopy revealed the notable absorption spectra at 425 nm with a wavelength shift around 450 nm, likely due to surface plasmon resonance excitation. SEM analysis showed spherical, monodisperse, nano scale particles with a size range of 50-60 nm. Crystaline phase of the synthesized nanoparticles was confirmed by x ray diffraction studies which showed the distinct peaks at (2θ) 27.90, 32.20, 46.30, 54.40, and 67.40, corresponding to (111), (200), (220), (222), and (311) planes of nano scale silver. The biocompatibility of these nanoparticles was assessed through zebrafish embryonic toxicity study which showed more than 90 % of embryos were alive and healthy. No marked changes on the blood cells also confirmed best hemocompatibility of the nanoparticles. Synthesized nanoparticles thus obtained were fabricated on the urinary catheter and the fabrication was confirmed by FTIR and SEM analysis. Notable changes in the absorption peaks, uniform coating and embedding of silver nanoparticles studied by FTIR and SEM analysis confirmed the fabrication of silver nanoparticles. The coated catheters demonstrated significant antibacterial activity against pathogenic bacterial strains, including E. coli ATCC 25922 and Pseudomonas aeruginosa ATCC 27853. Anti-biofilm studies, conducted using a modified microtiter plate crystal violet assay, revealed effective inhibition of both bacterial adhesion and biofilm development. 85 % of biofilm inhibition was recorded against both the tested strains. The coating method presented in this study shows promise for enhancing infection resistance in commonly used medical devices like urinary catheters, thus addressing device-associated infections., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Background: Since the onset of the coronavirus disease 2019 (COVID-19) pandemic, remarkable advances have been made in vaccine development to reduce mortality. However, therapeutic interventions for COVID-19 are comparatively limited despite these intensive efforts. Furthermore, the rapid mutation capability of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), a characteristic of its RNA structure, has led to the emergence of multiple variants, necessitating a shift from a predominantly vaccine-centric approach to one that encompasses therapeutic strategies. 6'-Hydroxy justicidin B (6'-HJB), an arylnaphthalene lignan isolated from Justicia procumbens, a traditional Chinese medicine, is known for its antiviral properties., Hypothesis/purpose: The aim of the present study was to assess the effectiveness and safety of 6'-HJB against SARS-CoV-2 in order to determine its potential as a therapeutic agent against COVID-19., Methods: The efficacy of 6'-HJB was evaluated both in vitro using Vero and Calu-3 cell lines and in vivo using ferrets. The safety assessment included toxicokinetics, safety pharmacology, and Good Laboratory Practice (GLP)-compliant toxicity evaluations following single- and repeated-dose toxicity studies in dogs., Results: The anti-SARS-CoV-2 efficacy of 6'-HJB was evaluated through dose-response curve (DRC) analysis using immunofluorescence; 6'-HJB demonstrated superior inhibition of SARS-CoV-2 growth and lower cytotoxicity than remdesivir. In SARS-CoV-2-infected ferret, 6'-HJB showed efficacy comparable to that of the positive control, Truvada. Further GLP toxicity studies corroborated the safety profile of 6'-HJB. Single-dose and 4-week repeated oral toxicity studies in Beagle dogs demonstrated minimal harmful effects at the highest dosages. The lethal dose of 6'-HJB exceeded 2,000 mg kg -1 in Beagle dogs. Toxicokinetic and GLP safety pharmacology studies demonstrated no adverse effects of 6'-HJB on metabolic processes, respiratory or central nervous systems, or cardiac functions., Conclusion: This research highlights both the antiviral efficacy and safety profile of 6'-HJB, underscoring its potential as a novel COVID-19 treatment option. The potential of 6'-HJB was demonstrated using modern scientific methodologies and standards., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier GmbH.. All rights reserved.)

This study delves into the antioxidant potential of Justicia gendarussa, commonly known as gendarussa, and its response to varying doses of nitrogen fertilizer. Gendarussa exhibits the potential for antioxidant activity. The diverse ecological conditions in which it thrives may influence its biological activity and lead to inconsistent production of secondary metabolites. Nitrogen, a pivotal factor in secondary metabolite production in plants, has become a focal point of this research. This research aims to determine the optimal nitrogen fertilizer dose on gendarussa antioxidant capacity and metabolites using a metabolomics approach. Employing a randomized block design for cultivation, the investigation revealed that a maximum harvest weight of 10.9 g/aerial parts of the plant was achieved with 270 kg/ha of nitrogen fertilizer. This study explored the DPPH, ABTS, FRAP, and CUPRAC assays of antioxidant effect, and found insignificant differences between the various nitrogen treatments. UHPLCQ-Orbitrap HRMS was employed to identify 30 metabolites in positive and 18 in negative ionization modes. Gendarusin A, a major metabolite in gendarussa, is identified in both positive and negative ionization. PCA and heatmap analysis successfully categorized these metabolites in the aerial parts of gendarussa at different nitrogen fertilizer dosages. Based on the metabolomics approach, variations in nitrogen fertilizer made metabolites at doses of 90 kg/ha had higher relative concentrations of metabolites compared to doses of 180 kg/ha and 270 kg/ha. So, 90 kg/ha are the optimal nitrogen fertilizer dose for cultivation and utilization strategies.

Ethnopharmacological Relevance: Justicia adhatoda L. is used as traditional medicine in Nepal to treat cough, asthma, and inflammatory disorders, and is indicated as "Asuro". Leaves are used worldwide as herbal medicine due to cardiotonic, expectorant, anti-asthmatic, and bronchodilatory properties. The aim of this work was to study the phytochemical composition of leaves of Nepalese J. adhatoda and assess their anti-inflammatory and antioxidant properties in vitro., Materials and Methods: Secondary metabolites were extracted from dried leaves using methanol (JAME: J. adhatoda methanol extract). They were analysed by means of liquid chromatography coupled with multiple-stage mass spectrometry (LC-MS n ). Anti-inflammatory potential was determined by the NF-κB and AP-1 inhibition assay, and DPPH, ABTS, and β-carotene bleaching assays were performed to assess its antioxidant properties., Results: JAME is a rich source of secondary metabolites, especially quinazoline alkaloids such as vasicine, vasicinone, vasicoline, and adhatodine. 7-Hydroxy derivatives of peganidine, vasicolinone, and adhatodine were also identified by means of MS n data and are here reported in J. adhatoda for the first time. JAME inhibited NF-κB and AP-1 expression in THP-1 cells to a greater extent than the positive control prednisolone. A moderate radical-quenching property was observed in DPPH and ABTS assays, but the anti-carotene bleaching activity was significantly higher than the reference BHT., Conclusions: To the best of our knowledge, this is the first insight into the phytochemical composition of Asuro leaves from Nepal and their bioactivity. Our results will contribute to the valorisation of this medicinal species still widely used in the traditional and complementary medicine., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Author(s). Published by Elsevier B.V. All rights reserved.)

The objective of the present study was to analyze and identify the phytochemical components found in neem leaf extracts using gas chromatography-mass spectrometry (GC-MS) and Fourier-transform infrared spectroscopy (FTIR) methods. The extract samples were acquired using ethyl acetate (EA) and petroleum ether (PE) solvents. Moreover, the extracts were assessed for their antibacterial and antioxidant features. In addition, chitosan nanoparticles (Cs NPs) containing neem extracts were synthesized and evaluated for their potential antibacterial properties, explicitly targeting multi-drug resistant (MDR) bacteria. The neem extracts were analyzed using GC-MS, which identified components such as hydrocarbons, phenolic compounds, terpenoids, alkaloids, and glycosides. Results revealed that the PE extract showed significant antibacterial activity against a range of bacteria. In addition, the PE extract exhibited significant antioxidant activity, exceeding both the EA extract and vitamin C. In addition, both extracts exhibited notable antibiofilm activity, significantly inhibiting the production of biofilm. The Cs NPs, loaded with neem extracts, exhibited significant antibacterial action against multidrug-resistant (MDR) microorganisms. The Cs NPs/EA materials had the greatest zone of inhibition values of 24 ± 2.95 mm against Pseudomonas aeruginosa. Similarly, the Cs NPs/PE materials exhibited a zone of inhibition measurement of 22 ± 3.14 mm against P. aeruginosa. This work highlights the various biochemical components of neem extracts, their strong abilities to combat bacteria and oxidative stress, and the possibility of Cs NPs containing neem extracts as effective treatments for antibiotic-resistant bacterial strains., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Introduction: Justicia pectoralis Jacq. is traditionally applied in folk medicine in Brazil and in several Latin American countries. The leaves are used in tea form, especially in the treatment of respiratory disorders, acting as an expectorant. It also has activity in gastrointestinal disorders, and it is anti-inflammatory, antioxidant, sedative, and estrogenic, among others., Aims: To investigate the gastroprotective activity of the methanol extract of the leaves of Justicia pectoralis Jacq. (MEJP) in different experimental models of gastric ulcers., Materials and Methods: The adult leaves of Justicia pectoralis Jacq. were collected and cultivated in beds, with an approximate spacing of 40 × 40 cm, organic fertilization, irrigation with potable water and without shelter from light. The MEJP was prepared from the dried and pulverized leaves and concentrated under reduced pressure in a rotary evaporator. For the experimental model of gastric ulcer, Swiss male albino mice were used. The inputs used in the experiment were MEJP at three different concentrations (250, 500 and 1000 mg/kg p.o.), cimetidine (50 mg/kg p.o.), indomethacin (50 mg/kg s.c.) and vehicle (10 mL/kg p.o.)., Results: MEJP (250, 500 and 1000 mg/kg p.o.) demonstrated gastroprotective activity, with levels of protection of 45.65%, 44.80% and 40.22%, respectively, compared to the control (vehicle). Compared with cimetidine (48.29%), MEJP showed similar gastroprotective activity., Conclusions: This study demonstrated the gastroprotective activity of MEJP and contributes to validate the traditional use the species for gastric disorders and provides a pharmacological basis for its clinical potential.

Vasicine from Adhatoda vasica was investigated in the management of aflatoxicosis and ochratoxicosis by in silico molecular docking approach. The computational analysis was carried out using Discovery Studio Autodock 4.5 tool. Absorption, distribution, metabolism, and excretion (ADME), pharmacodynamics and toxicity studies were also carried out using Swiss ADME and PASS online server, respectively. The standard drug compound used was silymarin and the structure were retrieved from the protein data bank for both the test compound vasicine and the standard drug. Vasicine interacted with aflatoxin B1 at 10 different poses and the maximum dock score was found to be 83.04 and the binding energy was -37.54 kcal/mol. Silymarin interacted with aflatoxin B1 at 10 different poses and the maximum dock score was found to be 143.578 and the binding energy was -67.32 kcal/mol. Vasicine interacted with ochratoxin A at 10 different poses and the maximum dock score was found to be 73.75 and the binding energy was -56.20 kcal/mol. Silymarin interacted with ochratoxin A at 10 different poses and the maximum dock score was found to be 89.23 and the binding energy was -98.86 kcal/mol. The compounds possess good gastro intestinal absorption with antioxidant property and exhibits minimum adverse effects. The obtained results support the toxin mitigating potential of the test compound with minimum adverse effects and hence vasicine can be regarded as a potential toxin binder of aflatoxin B1 and ochratoxin A, wherein it can be implemented for alleviating aflatoxicosis and ochratoxicosis., (Copyright © 2023 The Authors. Published by Elsevier Inc. All rights reserved.)

Ethnopharmacological Relevance: Justicia procumbens L. (JP) (Oriental Water Willow, Shrimp plant, Acanthaceae) is a herbaceous plant that is commonly found in India, Taiwan, Australia, Southern China, Vietnam, and Korea. The plant has been primarily used to treat fever, asthma, edema, cough, jaundice, urinary tract infection, and sore throat, as well as for snake bites and as a fish-killer. In the present review, the reported phyto-chemical, ethno-pharmacological, biological, and toxicological studies on J. procumbens were summarized. Special focus had been given to its reported lignans, regarding their isolation, characterization, quantitative estimation, and biosynthesis., Materials and Methods: A survey of the literature was done using assorted databases and publishers; Scopus, Sci-Finder, Web of Science, PubMed, GoogleScholar, ScienceDirect, Wiley, Taylors&Francis, Bentham, Thieme, and Springer., Results: Currently, 95 metabolites have been separated fromJ. procumbens. Lignans and their glycosides were reported as main phyto-constituents of J. procumbens. Various methods are mentioned for quantitative estimation of these lignans. These phyto-constituents possessed wide pharmacological effectiveness, such as antiplatelet aggregation, antimicrobial, antitumor, and antiviral., Conclusions: Many of the stated effects are harmonious with the reported traditional uses of this plant. This data could further support J. procumbens's utilization as a herbal remedy and drug lead. However, further study of J. procumbens toxicity, as well as preclinical and clinical investigation is required to ensure the safe usage of J. procumbens., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Objectives: Antibiotic resistance is rising, prompting innovative strategies for eradicating the epidemic. This study investigated the antibacterial properties of the leaves of a widely used medicinal plant, Adhatoda vasica ., Methods: The plant's polar (water, methanol) and non-polar (hexane) extracts were tested against several different bacterial strains using the disc diffusion technique., Results: In a study, it was found that the water extract had the greatest inhibitory effect on Staphylococcus simulans and Staphylococcus aureus , with minimum inhibitory concentrations of 16.444 and 19.315 g/mL, respectively. Gram-negative strains were more susceptible to plant extracts than Gram-positive strains. The phytochemical analysis indicated the presence of secondary metabolites such as alkaloids, saponins, flavonoids, tannins, and steroids, where absorbance was recorded at 415 nm. The water extract had the highest amount of phenolics, with a total phenolic content of 53.92 0.47 mg and a total flavonoid content of 7.25 0.08 mg. Results suggest that the extract may have potential therapeutic applications for antimicrobial properties., Conclusions: The study concluded that the extract's phenolic group of secondary metabolites were responsible for its antibacterial activity. The study highlights A. vasica as a promising source for discovering new and effective antibacterial compounds., (© 2023 Walter de Gruyter GmbH, Berlin/Boston.)

The genus Justicia has more than 600 species distributed in both hemispheres, in the tropics and temperate regions, and it is used in the treatment of numerous pathologies. This study presents a review of the biological activities of plant extracts and isolated chemical constituents of Justicia (ACANTHACEAE), identified in the period from May 2011 to August 2022. We analyzed over 176 articles with various biological activities and chemical compound descriptions present in the 29 species of Justicia . These have a variety of applications, such as antioxidant and antimicrobial, with alkaloids and flavonoids (e.g., naringenin) the most frequently identified secondary metabolites. The most observed species were Justicia gendarussa Burm., Justicia procumbens L., Justicia adhatoda L., Justicia spicigera Schltdl, and Justicia pectoralis Jacq. The frontier molecular orbitals carried out using density functional theory (M062X and basis set 6-311++G(d,p) indicate reactive sites for naringenin compound and a chemical reaction on phytomedicine activity. The energy gap (206.99 kcal/mol) and dimer solid state packing point to chemical stability. Due to the wide variety of pharmacological uses of these species, this review points toward the development of new phytomedicines.

Objective: The present study is an in silico model of platelet amplification potential of Adhatoda vasica, which can be used to treat thrombocytopenia in dengue complications., Methods: Docking studies have proved to be an essential tool that facilitates the structural diversity of natural products to be harnessed in an organized manner. In the present study, vasicine containing natural anti-dengue potential was subjected to docking studies using Schrodinger glides software (ver.11.1). The docking study was carried out to find out the potential molecular targets for selected protein. The docking was carried out on different ligands, like vasicine, ramatroban, chloroquine, celgosivir, and standard eltrombopag downloaded from PubChem and retrieved to glide software and ligands prepared using lig prep wizard. Docking was performed using the ligand docking wizard of Glide-maestro 2018., Results: The docking score of vasicine (-5.27) is nearly identical to the standard eltrombopag (-6.08), and both ligands bind with one hydrogen bond. The validation score of ramatroban is -12.39, binding with five hydrogen bonds, Celgosivir exhibited a docking score of -7.3 with three hydrogen bonds, and chloroquine displayed no hydrogen bond but had a docking score of -4.6., Conclusion: Vasicine was found to be the most suitable target of platelet amplification potential from Adhatoda vasica. However, the molecular docking results are preliminary, and it has been indicated that vasicine could be one of the potential ligands to treat the thrombocytopenia of dengue; experimental evaluation will be carried out in the near future., (Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.net.)

Eleven previously undescribed arylnaphthalide lignans ( 1 - 11 ) together with seven known compounds were isolated from the whole plant of Justicia depauperata . The structures of 1 - 11 were elucidated by spectroscopic analysis and mass spectrometry. Compounds 6 (IC 50 = 4.1 μM) and 9 (IC 50 = 9.5 μM) displayed cytotoxic activity against the KB-3-1 cervical carcinoma cell line. This report provides an insight into the conformational equilibria occurring in the arylnaphthalide lignan constituents of this plant.

Natural products are being targeted as alternative anticancer agents due to their non-toxic and safe nature. The present study was conducted to explore the in vitro anticancer potential of Justicia adhatoda (J. adhatoda) leaf extract. The methanolic leaf extract was prepared, and the phytochemicals and antioxidant potential were determined by LCMS analysis and DPPH radical scavenging assay, respectively. A docking study performed with five major alkaloidal phytoconstituents showed that they had a good binding affinity towards the active site of NF-κB. Cell viability assay was carried out in five different cell lines, and the extract exhibited the highest cytotoxicity in MCF-7, a breast cancer cell line. Extract-treated cells showed a significant increase in nitric oxide and reactive oxygen species production. Cell cycle analysis showed an arrest in cell growth at the Sub-G0 phase. The extract successfully inhibited cell migration and colony formation and altered mitochondrial membrane potential. The activities of superoxide dismutase and glutathione were also found to decrease in a dose-dependent manner. The percentage of apoptotic cells was found to increase in a dose-dependent manner in MCF-7 cells. The expressions of caspase-3, Bax, and cleaved-PARP were increased in extract-treated cells. An increase in the expression of NF-κB was found in the cytoplasm in extract-treated cells. J. adhatoda leaf extract showed a potential anticancer effect in MCF-7 cells.

Chiral compounds find importance as drugs and therapeutic targets. Enantiomers of chiral drugs have been found to show different biological properties like pharmacokinetics, toxicology, pharmacology, metabolism, and so forth. In this study, we have identified the chiral compounds present in the medicinal plant Adhatoda vasica Nees (Justicia adhatoda Linn). Phytochemical investigation on the leaves of Justicia adhatoda resulted in the identification of 27 chiral compounds. We report diverse compounds identified in the crude methanolic extract of Justicia adhatoda leaves by GC-MS analysis exhibiting diverse biological activities. Quantitative analysis of anticancer compound dihydroxycolchicine from the methanolic extract of J. adhatoda leaves was done by external standard method, and the amount of anticancer compound dihydroxycolchicine was found to be 87.823 mg/l indicative of moderate production in the leaves. Therefore, the extract of leaves of Justicia adhatoda can be used as a potential source of chiral bioactive molecules of pharmacological importance for drug synthesis., (© 2022 Wiley Periodicals LLC.)

Plants are regarded as a valuable and inexpensive source of new drug development, and a variety of plant compounds are now being used in clinical trials to treat a variety of ailments. The goal of this work was to characterize and evaluate the anti-inflammatory and antioxidant effects of Justicia adhatoda L. leaf extract (Acanthaceae). The presence of alkaloids, saponins, tannins, phytosterols, phenols, and proteins in the leaf extract of J. adhatoda was determined using phytochemical screening. While the identification of different compounds in the leaf extract was carried out by HPLC analysis. Similarly, the anti-inflammatory potential of the leaf extract was assessed in Carrageenan and Formalin-induced inflammatory mice models. The phytochemical analysis of the leaf extract indicated a positive test for alkaloids, saponins, tannins, phytosterols, phenols, proteins, and amino acids, while the negative test for carbohydrates, and glycosides, flavonoids, and diterpenes. Moreover, among the detected compounds, gallic acid was found in the highest concentration with a 45.42% composition. The leaf extract showed the highest antibacterial activity against E. coli, while the lowest activity against Listeria was observed. The leaf extract of J. adhatoda revealed promising anti-inflammatory, analgesic, and antioxidants activities both in vitro and in vivo. Similarly, the detected compounds portrayed variable pharmacokinetic as well as binding affinities with the target proteins. In conclusion, the leaf extract exhibited significant antioxidants and antibacterial activities using in vitro assays. Similarly, the extract also revealed promising anti-inflammatory activities in vivo while exhibiting variable Pharmacokinetics and binding affinities towards protein target using computational tools., Competing Interests: Conflict of interest statement The authors declare no conflict of interest., (Copyright © 2022 The Authors. Published by Elsevier Masson SAS.. All rights reserved.)

Ethnopharmacological Relevance: Justicia procumbens L. is a traditional Chinese medicine, first recorded in "Shen Nong's Herbal Classic", for the treatment of lumbar pain and fever. As a widely distributed herb, it has also been documented in India, Nepal, and Malaysia. In "Tang Materia Medica", a famous medicinal book of Tang Dynasty in ancient China, it was first used to treat diseases associated with blood stasis. Blood stasis syndrome is closely related to thrombus formation and platelet aggregation. Although some compounds isolated from this plant have anti-platelet aggregation effects, the main chemical components and mechanism of J. procumbens in terms of these effects are little known., Aims of the Study: Through in vivo and in vitro experiments, this studsy revealed the characteristic components and action mechanism of anti-platelet aggregation by J. procumbens from an overall perspective., Materials and Methods: The effective crude extracts of the whole plant were screened via an in vitro anti-platelet aggregation test. After incubating these extracts with apheresis platelets, high affinity compounds were detected by HPLC-MS and regulatory genes were detected using gene chips. The effective components and potential target proteins were analyzed using computational docking technology. Furthermore, the compound with the strongest predicted activity was evaluated in vivo via an anti-thrombotic test., Results: Integrin a Ⅱb β 3 , PKCα, PI3Kγ, and mitogen-activated protein kinase 14 were found to be potential targets. Justicidin B, tuberculatin, chinensinaphthol methyl ether, and neojusticin B were effective compounds that inhibited human platelet aggregation by suppressing Gq-PLC-PKC and Gi-PI3K-MAPK signaling pathways. Among the compounds that bind to platelets, justicidin B showed the strongest virtual binding force. The test of carotid artery thrombosis induced by ferric chloride in SD rats confirmed that justicidin B inhibited thrombus formation., Conclusion: Experimental investigation showed that arylnaphthalene lignan aglycones with one methylenedioxy group and two methoxy groups are effective components for anti-platelet aggregation by J. procumbens. These compounds inhibit Gq-PLC-PKC and Gi-PI3K-MAPK signaling pathways by suppressing the expression of genes such as ITGB3, PRKCA, PIK3CG, and MAPK14. These results reflected the characteristics of multi-component and multi-target synergistic treatment of Chinese medicine., (Copyright © 2022 Elsevier B.V. All rights reserved.)

This article reports the three principal groups of compounds for the first time from Adhatoda vasica and Calotropis procera plants species using nuclear magnetic resonance methods in which aliphatic, oxy heterocyclic, and tannins compounds were detected from these plants. The leaves of both species were subjected to testing tyrosinase inhibition and antioxidant activities. ATP bioluminescence use for indirect measurement of the amount of organic residue on the surface of the leaves that provide support to microbial growth. The distinguishing characteristics and intraoperative findings of bacterial diseases involved in treatments were conducted against the positive and negative microbial strains using a scanning electron microscope (SEM). The methanolic extracts of leaves of both species were applied to bacterial strains through broth microdilution method to determine the minimum inhabitation concentrations (MICs) for both species. It was concluded that both plants are a rich resource of bioactive compounds. Their extract may also be used to treat various bacterial diseases and in drug manufacturing. HIGHLIGHTS: New chemical compounds of oxy-heterocyclic, aliphatic, and tannins derivatives are isolated from herbal plants as a source of various drugs. 1 H NMR spectrum and 13 C NMR spectrum of each new derivate were calculated. NMR-spectral analysis of new compound of chemistry class was studied and further applied in various bacterial strains. Tyrosinase inhibition property of bacteria strains by application of active compounds on these strains. Agar overlay bioassays were used to evaluate intercellular morphological features of strains applied on extracts by electron microscope (SEM). a-Glucosidase inhibition assay determined with antioxidants activity through FRAP assay methods., (© 2021 Wiley Periodicals LLC.)

Ethnopharmacological Relevance: Justicia vahlii Roth. (Acanthaceae), also called as kodasoori and bhekkar is an annual therophyte erect or decumbent herb used traditionally in toothache, skin diseases (itching, topical inflammation) and for the treatment of various respiratory disorders., Aim of the Study: The current study aimed at exploring pain cessation potential of J. vahlii Roth. via murine model of neuropathic pain and its phytochemical, toxicological and antioxidant profiles., Materials and Methods: The hydro-alcoholic extract of J. vahlii (HAEJv) prepared by maceration technique was subjected to preliminary phytochemical screening, total bioactive content determination, UPLC-QTOF-MS and GC-MS analysis. Toxicity assessment was carried out by using brine shrimp lethality assay and acute oral toxicity test. Murine model of neuropathic pain was applied to assess the antineuropathic potential of the species. Furthermore effect of the extract on catalase, superoxide oxide dismutase (SOD), Glutathione (GSH), interleukin-1beta (IL-1β) and total necrosis factor-alpha (TNF-α) was also studied. In vitro antioxidant profile was explored by using four methods; 2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2-azinobis(3-ethylbenothiazoline)-6-sulfonic acid (ABTS), CUPric reducing antioxidant capacity (CUPRAC) and Ferric reducing antioxidant power (FRAP) assay., Results: The phytochemical screening revealed the presence of phenols, flavonoids, coumarins, alkaloids and lignans as the major classes of secondary metabolites. The extract was found rich in total phenolics content (TPC) and total flavonoids content (TFC) with identification of total 59 bioactives in UPLC-QTOF-MS and 40 compounds in GC-MS analysis. The extract was found nontoxic up to 4000 mg/kg (p.o.) in mice and no mortality observed in brine shrimp lethality assay. The HAEJv significantly reduced number of acetic acid induced abdominal constrictions at 100 mg/kg (p < 0.01) and 200 mg/kg (p < 0.001) and increased paw withdrawal threshold p < 0.05 at 100 mg/kg and p < 0.001 at 200 mg/kg, and an increase in tail withdrawal latency time p < 0.001 at 200 mg/kg was observed. The extract significantly increased levels of catalase, SOD and GSH while decreased IL-1β and TNF-α levels in sciatic nerve tissue of mice. HAEJv showed highest antioxidant activity through CUPRAC method 121.32 ± 1.22 mg trolox equivalent per gram of dry extract (mg TE/g DE) followed by DPPH 81.334 ± 4.35 mg TE/g DE, FRAP 69.89 ± 3.05 mg TE/g DE and ABTS 38.17 ± 2.12 mg TE/g DE., Conclusion: The current study back the traditional use of J. vahlii in pain cessation through antioxidant based antineuropathic pain activity and revealed the extract non-toxic with number of functional phytoconstituents and warrants further research on isolation of the compounds and sub-acute toxicity studies., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Inflammation plays a major role in the onset and progression of many diseases related to the respiratory system. Cysteinyl leukotrienes, the products of 5-LOX are a potent bronchoconstrictor. Vasicine, vasicinone and deoxyvasicine are the pyrroquinazoline alkaloids of Adhatoda vasica that are well known for their bronchodilatory activity. The current investigation evaluates the 5-LOX inhibitory potential of these alkaloids. Molecular docking results indicated that these alkaloids have similar binding energy as that of Zileuton, a commercial drug. Analysis of the molecular dynamics simulations, the binding free energy derived from MM-PBSA and interaction entropy indicated that vasicinone (-8.33 kcal/mol) exhibited a binding free energy comparable to that of Zileuton (-8.52 kcal/mol). The in-vitro results indicate the potential of vasicinone as a competitive inhibitor, while the in-silico results highlighted the potential of vasicine and deoxyvasicine as allosteric inhibitors. A possible mechanism behind the activity exhibited by the plant was also determined, which emphasized the potential of these alkaloids as leads for the design of novel 5-LOX inhibitors.

Angiotensin converting enzyme (ACE) plays a crucial role in regulating blood pressure in the human body. Identification of potential ACE inhibitors from medicinal plants supported the idea of repurposing these medicinal plants against hypertension. A method based on ultra-performance liquid chromatography (UPLC) coupled with a diode array detector (DAD) was used for the rapid screening of plant extracts and purified compounds to determine their ACE inhibitory activity. Hippuryl-histidiyl-leucine (HHL) was used as a substrate, which is converted into hippuric acid (HA) by the action of ACE. A calibration curve of the substrate HHL was developed with the linear regression 0.999. The limits of detection and quantification of this method were found to be 0.134 and 0.4061 mM, respectively. Different parameters of ACE inhibitory assay were optimized, including concentration, incubation time and temperature. The ACE inhibition potential of Adhatoda vasica (methanolic-aqueous extract) and its isolated pyrroquinazoline alkaloids, vasicinol ( 1 ), vasicine ( 2 ) and vasicinone ( 3 ) was evaluated. Compounds 1 - 3 were characterized by various spectroscopic techniques. The IC 50 values of vasicinol ( 1 ), vasicine ( 2 ) and vasicinone ( 3 ) were found to be 6.45, 2.60 and 13.49 mM, respectively. Molecular docking studies of compounds 1 - 3 were also performed. Among these compounds, vasicinol ( 1 ) binds as effectively as captopril, a standard drug of ACE inhibition., Competing Interests: The authors declare no conflict of interest.

Three novel 14-membered cyclopeptide alkaloids, justicianenes B-D ( 1 - 3 ), were isolated from the EtOH extract of the whole plant of Justicia procumbens L., and their structures were determined on the basis of detailed NMR spectroscopic data and the absolute stereochemistry of the ring-bonded α-amino acids in the cyclopeptide alkaloids were determined by ECD spectra. The isolated compounds were evaluated for their in vitro cytotoxicity against human cancer cell lines, including brest cancer MCF-7, cervix carcinoma HeLa, lung cancer A549 and H460, and diphyllin ( 14 ) showed moderate cytotoxicity against the HeLa, A549 and H460 cells with IC 50 of 9.13, 23.12, 42.34 µM, respectively, justicianene D showed weak cytotoxicity against the MCF-7 cell with inhibition rate of 50% at the concentration of 90 µM.

Objectives: The toxicity of chemotherapeutic anticancer drugs is a serious issue in clinics. Drug discovery from edible and medicinal plants represents a promising approach towards finding safer anticancer therapeutics. Justicia insularis T. Anderson (Acanthaceae) is an edible and medicinal plant in Nigeria. This study aims to discover cytotoxic compounds from this rarely explored J. insularis and investigate their underlying mechanism of action., Methods: The cytotoxicity of the plant extract was evaluated in human ovarian cancer cell lines and normal human ovarian surface epithelia (HOE) cells using a sulforhodamine B assay. Bioassay-guided isolation was carried out using column chromatography including HPLC, and the isolated natural products were characterized using GC-MS, LC-HRMS, and 1D/2D NMR techniques. Induction of apoptosis was evaluated using Caspase 3/7, 8, and 9, and Annexin V and PI based flow cytometry assays. SwissADME and SwissTargetPrediction web tools were used to predict the molecular properties and possible protein targets of identified active compounds. Key finding: The two cytotoxic compounds were identified as clerodane diterpenoids: 16(α/β)-hydroxy-cleroda-3,13(14) Z -dien-15,16-olide ( 1 ) and 16-oxo-cleroda-3,13(14) E -dien-15-oic acid ( 2 ) from the Acanthaceous plant for the first time. Compound 1 was a very abundant compound (0.7% per dry weight of plant material) and was shown to be more potent than compound 2 with IC 50 values in the micromolar range against OVCAR-4 and OVCAR-8 cancer cells. Compounds 1 and 2 were less cytotoxic to HOE cell line. Both compounds induced apoptosis by increasing caspase 3/7 activities in a concentration dependent manner. Compound 1 further increased caspase 8 and 9 activities and apoptosis cell populations. Compounds 1 and 2 are both drug like, and compound 1 may target various proteins including a kinase., Conclusions: Clerodane diterpenoids ( 1 and 2 ) in J. insularis were identified as cytotoxic to ovarian cancer cells via the induction of apoptosis, providing an abundant and valuable source of hit compounds for the treatment of ovarian cancer.

Thrombotic diseases seriously threaten human life. Justicia, as a common Chinese medicine, is usually used for anti-inflammatory treatment, and further studies have found that it has an inhibitory effect on platelet aggregation. Therefore, it can be inferred that Justicia can be used as a therapeutic drug for thrombosis. This work aims to reveal the pharmacological mechanism of the anti-thrombotic effect of Justicia through network pharmacology combined with wet experimental verification. During the analysis, 461 compound targets were predicted from various databases and 881 thrombus-related targets were collected. Then, herb-compound-target network and protein-protein interaction network of disease and prediction targets were constructed and cluster analysis was applied to further explore the connection between the targets. In addition, Gene Ontology (GO) and pathway (KEGG) enrichment were used to further determine the association between target proteins and diseases. Finally, the expression of hub target proteins of the core component and the anti-thrombotic effect of Justicia's core compounds were verified by experiments. In conclusion, the core bioactive components, especially justicidin D, can reduce thrombosis by regulating F2, MMP9, CXCL12, MET, RAC1, PDE5A, and ABCB1. The combination of network pharmacology and the experimental research strategies proposed in this paper provides a comprehensive method for systematically exploring the therapeutic mechanism of multi-component medicine., (© 2021. The Author(s).)

Obesity is a global health problem characterized by excessive fat deposition in adipose tissues and can be managed by targeting pancreatic lipase (PL) activity. In the present study, we investigated the in vitro antioxidant and anti-obesity potentials of methanolic leaf extract of Justicia carnea(MEJC) using lipase inhibition kinetics model. In silico evaluations of MEJC bioactive compounds as potential drug-like agents and inhibitors of PL were also investigated using SwissADME prediction tool, semi-empirical quantum mechanics(SQM), molecular electrostatic potential(MEP) and molecular docking analysis. Gas chromatography-mass spectrometry(GC-MS) revealed presence of campesterol, stigmasterol, beta-amyrin etc. MEJC scavenged reactive species and inhibited PL activity via a mixed inhibition pattern (K i  = 107.69 μg/mL; K ii  = 398.00 μg/mL) with IC 50  > orlistat's IC 50 . Molecular docking of GC-MS identified compounds with porcine PL showed compounds 8,10,12 and 14 having high PL-binding affinity and similar binding pose with orlistat. Hydrophobic interactions and van der Waals forces were predominantly involved in the ligands' interactions with some key catalytic site amino acid residues (Ser-153,His-264). Compounds 10,12,13 and 14 indicated high drug-likeness, bioavailability, electronegativity, E LUMO -E HOMO energy gaps and MEP. Our findings show that MEJC is a rich natural source of antioxidant and anti-obesity agents which could be optimized for development of new anti-obesity drugs., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Justicia spicigera Schltdl. (Acanthaceae) is used for treatment of gastrointestinal illnesses therapy in traditional medicine. The objective of this study was to give evidence of the antinociceptive and spasmolytic effects of the J. spicigera ethanol extract (JS EtOH) using in in vivo and/or in vitro assays. The JS EtOH exerted regulatory effect on the motility and a partial relaxing response on the intestinal tissue. Furthermore, a significant abdominal antinociceptive response was obtained in mice, which was totally abolished in the presence of 5-HT 1A receptor antagonist (WAY100635, 0.1 mg/kg, s.c.) and partially by blocking opioid receptors (NX, 1 mg/kg, i.p.), whereas the inhibition of the NO synthesis (L-NAME, 30 mg/kg, i.p.) facilitated antinociception of this extract. Kaempferitrin was isolated and identified as major secondary metabolite. These results support the analgesic and spasmolytic-like activity of J. spicigera aerial parts involving inhibitory neurotransmission reinforcing the potential of this medicinal plant for alleviating pain.

Severe asthma is a chronic airway disease that exhibits poor response to conventional asthma therapies. Growing evidence suggests that elevated hypoxia increases the severity of asthmatic inflammation among patients and in model systems. In this study, we elucidate the therapeutic effects and mechanistic basis of Adhatoda vasica (AV) aqueous extract on mouse models of acute allergic as well as severe asthma subtypes at physiological, histopathological, and molecular levels. Oral administration of AV extract attenuates the increased airway resistance and inflammation in acute allergic asthmatic mice and alleviates the molecular signatures of steroid (dexamethasone) resistance like IL-17A, KC (murine IL-8 homologue), and HIF-1α (hypoxia-inducible factor-1α) in severe asthmatic mice. AV inhibits HIF-1α levels through restoration of expression of its negative regulator-PHD2 (prolyl hydroxylase domain-2). Alleviation of hypoxic response mediated by AV is further confirmed in the acute and severe asthma model. AV reverses cellular hypoxia-induced mitochondrial dysfunction in human bronchial epithelial cells-evident from bioenergetic profiles and morphological analysis of mitochondria. In silico docking of AV constituents reveal higher negative binding affinity for C and O-glycosides for HIF-1α, IL-6, Janus kinase 1/3, TNF-α, and TGF-β-key players of hypoxia inflammation. This study for the first time provides a molecular basis of action and effect of AV whole extract that is widely used in Ayurveda practice for diverse respiratory ailments. Further, through its effect on hypoxia-induced mitochondrial dysfunction, the study highlights its potential to treat severe steroid-resistant asthma.

Ethnopharmacological Relevance: Adhatoda vasica Nees is widely used herb of indigenous system to treat various ailments especially upper respiratory tract infections. Not only, anti-tubercular efficacy of crude extract and phytoconstituents of A. vasica has been documented but its hepatoprotective role against various drugs mediated hepatic alterations in different animal models has also been observed., Background and Purpose: Isoniazid, rifampicin and pyrazinamide (H-R-Z) are anti-tubercular drugs normally prescribed by health professionals for the treatment of tuberculosis, however along with their medical effectiveness these drugs also exhibit hepatotoxicity among TB patients. Unexpectedly, substantial toxicological data on the metabolism of anti-TB drugs are available but the mystery behind these xenobiotics is too complex and partly implicit. In this study, we further explored the hepatotoxic effects of these xeno-metabolic products and their amelioration by Adhatoda vasica Nees by elucidating its mechanistic action., Methods: We generated a hepatotoxic rodent model by oral administration of H, R and Z (30.85, 61.7 and 132.65 mg/kg body weight) drugs for 25 days in Wistar rats. Additionally, to achieve hepatoprotection two different doses of Adhatoda vasica Nees ethanolic leaf extract (200 and 300 mg/kg body weight) were used along with H-R-Z dosage, orally and once daily for 25 days and tried to ascertain their mechanistic action. For this, initially phytoconstituents of the extract were evaluated followed by extract standardization using RP-HPLC and FTIR methods. Furthermore, antioxidant activity of the extract was analyzed by DPPH assay. Finally, different treated groups were analyzed for hepatic oxidative stress markers, antioxidant markers, histopathological changes and gene expression study including CYP2E1, CYP7A1, NAT, NR1I2 and UGT1A1 genes involved in phase I and phase II xeno-metabolism., Results: Estimated content of vasicine in RP-HPLC method and free-radical scavenging activity in DPPH assay was found to be 134.519 ± 0.00269μg/10mg of leaf extract and 47.81 μg/mL respectively. In H-R-Z treated group, a significant increase in the levels of thiobarbituric acid, significant reduction in the levels of GSH, and enzymatic markers and marked changes in hepatic histological architecture were observed. In addition, there was significance up-regulation of CYP7A and NAT genes, down-regulation of CYP2E1 gene and insignificant expression levels of NR1I2 and UGT1A1 genes were observed in H-R-Z group. Conversely, high dose of A. vasica extract effectively diminished these alterations by declining oxidative stress and boosting of antioxidant levels. In addition, it acted as bi-functional inducer of both phase I (CYP2E1) and phase II (NAT and UGT1A1) enzyme systems., Conclusion: Hence, we concluded that anti-TB drugs exposure has potential to generate reactive metabolites that eventually cause hepatotoxicity by altering oxidant-antioxidant levels and their own metabolism. This study not only emphasized on xeno-metabolism mediated hepatic alterations but also explore the benefit of A. vasica on these toxic insults., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Background: COVID-19 pneumonia has been associated with severe acute hypoxia, sepsis-like states, thrombosis and chronic sequelae including persisting hypoxia and fibrosis. The molecular hypoxia response pathway has been associated with such pathologies and our recent observations on anti-hypoxic and anti-inflammatory effects of whole aqueous extract of Adhatoda Vasica (AV) prompted us to explore its effects on relevant preclinical mouse models., Methods: In this study, we tested the effect of whole aqueous extract of AV, in murine models of bleomycin induced pulmonary fibrosis, Cecum Ligation and Puncture (CLP) induced sepsis, and siRNA induced hypoxia-thrombosis phenotype. The effect on lung of AV treated naïve mice was also studied at transcriptome level. We also determined if the extract may have any effect on SARS-CoV2 replication., Results: Oral administration AV extract attenuates increased airway inflammation, levels of transforming growth factor-β1 (TGF-β1), IL-6, HIF-1α and improves the overall survival rates of mice in the models of pulmonary fibrosis and sepsis and rescues the siRNA induced inflammation and associated blood coagulation phenotypes in mice. We observed downregulation of hypoxia, inflammation, TGF-β1, and angiogenesis genes and upregulation of adaptive immunity-related genes in the lung transcriptome. AV treatment also reduced the viral load in Vero cells infected with SARS-CoV2., Conclusion: Our results provide a scientific rationale for this ayurvedic herbal medicine in ameliorating the hypoxia-hyperinflammation features and highlights the repurposing potential of AV in COVID-19-like conditions.

Ethnopharmacological Relevance: Adhatoda vasica Nees., which existed in a large; number of Tibetan medicine prescriptions for hepatopathy, used as an adjuvant to treat liver diseases., Hypothesis/purpose: Oxidative stress is the key player in the development and progression of liver pathogenesis. In recent years, research is increasingly being focused on exploitation of the active components from medicinal plants to combat the liver oxidative injury. In our study, we aimed to screen the active principles from A. vasica and clarify whether they could relieve oxidative damage induced by tert-Butyl hydroperoxide (t-BHP) and its potential mechanism via activating AMPK/p62/Nrf2 pathway., Materials and Methods: Ultra performance liquid chromatography (UPLC) was adopted for analysis of chemical composition in the extracts. Furthermore, the antioxidant activity of the fractions was evaluated using DPPH, ABTS and reducing power assay. Along with this, the compounds in this fraction with highest antioxidant activity were analyzed using UPLC-MS. Based on this, the condition for extracting flavonoids of this subfraction was optimized via response surface method. CCK-8 assay was used to detect cell viability. Detection kits were used to measure the activity changes of AST, ALT, LDH and CAT as well as MDA and GSH levels induced by t-BHP. Detection of reactive oxygen species (ROS) production was used DCFH-DA probe. DAPI staining and flow cytometry was used to detect cell apoptosis. In terms of the mechanistic studies, the expression of proteins involved in AMPK/p62/Nrf2 pathway was measured using western blotting., Results: Eventually, 70% ethanol extract from leaf of A. vasica was chosen due to its highest active components compared with other extracts. Further, ethyl acetate fraction derived from 70% ethanol extract in A. vasica (AVEA) possess highest ability for scavenging DPPH and ABTS free radicals as well as strongest reducing power than other fractions. Chemical composition analysis showed that AVEA contained 17 compounds, including 1 quinazoline alkaloid, 12 flavonoid-C-glycosides and 4 flavonoid-O-glycosides. In addition, the conditions (ratio of solid-liquid 1:14, the concentration of ethanol 73%, and the temperature 65 °C) were selected to enrich the flavonoids in AVEA. Furthermore, AVEA could attenuate t-BHP induced hepatocyte damage via increasing the cell viability, restoring abnormal the activities of AST, ALT, LDH and CAT as well as the levels of MDA and GSH. ROS fluorescence intensity was reduced by AVEA. Meanwhile, it could inhibit the cell apoptosis of BRL 3 A cells, as evidenced by restoration of cell morphology and decreasing the number of apoptotic cells. Further mechanistic studies indicated AVEA could promote p-AMPK expression to further induce autophagy adaptor-p62 protein expression, which could autophagic degradation of Keap1, leading to Nrf2 release and translocation into nucleus to induce antioxidant genes (HO-1, NQO-1, GCLC and GCLM) expression., Conclusion: In our study, AVEA was first to screen as the active fraction in A. vasica with alkaloids and abundant flavones. Moreover, the fraction potentiates its beneficial aspect by displaying the protective role on relieving t-BHP induced oxidative stress and activating AMPK/p62/Nrf2 pathway. AVEA helps maintain the redox homeostasis of hepatic cells and could be considered as an effective candidate against oxidative stress related liver disorders., (Copyright © 2020 Elsevier B.V. All rights reserved.)

The present study investigated the antioxidant and invitro antidiabetic capacities of Justicia carnea aqueous leaf extract (JCAE) using α-amylase inhibition model. α-Amylase binding-interaction with JCAE was also investigated using fluorescence spectroscopy and molecular docking. Phytochemical screening and Gas Chromatography-Mass Spectrometry (GC-MS) analysis indicated presence of bioactive compounds. Phenolic (132 mg GAE/g) and flavonoid contents (31.08 mg CE/g) were high. JCAE exhibited high antioxidant capacity and effectively inhibited α-amylase activity (IC 50 , 671.43 ± 1.88 μg/mL), though lesser than acarbose effect (IC 50 , 108.91 ± 0.61 μg/mL). α-Amylase intrinsic fluorescence was quenched in the presence of JCAE. Ultraviolet-visible and FT-IR spectroscopies affirmed mild changes in α-amylase conformation. Synchronous fluorescence analysis indicated alterations in the microenvironments of tryptophan residues near α-amylase active site. Molecular docking affirmed non-polar interactions of compounds 6 and 7 in JCAE with Asp-197 and Trp-58 residues of α-amylase, respectively. Overall, JCAE indicated potential to prevent postprandial hyperglycemia by slowing down carbohydrate hydrolysis., (Copyright © 2020 Elsevier B.V. All rights reserved.)

In this work, the antibacterial activity of a crude extract of the endophytic fungus Flavodon flavus (JB257), isolated from leaves of Justicia brandegeana, was evaluated against both the vegetative and sporulated forms of Alicyclobacillus acidoterrestris. The microdilution technique was performed in order to determine the antibacterial activity of the crude extract alone as well as in combination with the bacteriocin, nisin. The minimum inhibitory concentration (MIC) of the crude extract and nisin alone against A. acidoterrestris vegetative forms were 250 μg/mL and 31.5 μg/mL, respectively, while the minimum bactericidal concentrations (MBC) were 1000 μg/mL and 62.5 μg/mL,respectively. For A. acidoterrestris spores, treatment with the crude extract at a concentration of 500 μg/mL caused a 47% reduction in growth, while nisin at 62.5 μg/mL could reduce 100% of the growth. The in vitro evaluation of the crude extract combined with nisin against A. acidoterrestris by the Checkerboard method showed a synergistic interaction between the two compounds. In addition, greater selectivity towards bacterial cells over host cells, a human hepatocyte cell line, was achieved when the crude extract was combined with nisin, Using scanning electron microscopy, interferences in the cell membrane of A. acidoterrestris could be observed after treatment with the crude extract. The results presented in this study indicate that the crude extract of the endophyte F. flavus has biotechnological potential in the food industry, especially for the treatment of orange juices through the control of A. acidoterrestris., (Copyright © 2020 Elsevier B.V. All rights reserved.)

New Findings: What is the central question of this study? Can Justicia flava leaf extract (JF) inhibit human myometrial contractility as was previously shown in mouse myometrium? What is the main finding and its importance? JF abolished human myometrial contractions and therefore presents as a lead plant in drug discovery studies involving drugs for preterm birth., Abstract: In the search for new potent therapies for preterm labour, Justicia flava leaf extract (JF) was previously shown to potently inhibit uterine contractility in both pregnant and non-pregnant mouse uterus. This study took the investigation a step further and investigated the activity of JF on pregnant human myometrial contractility. JF potently inhibited human myometrial contractility in a concentration-dependent manner. This pilot study provides evidence that JF should be further investigated as a lead plant in the drug discovery of new uterine relaxants., (© 2020 The Authors. Experimental Physiology © 2020 The Physiological Society.)

Justicia adhatoda L. (Acanthaceae), an Old-World species of Justicia, is found in almost all geographical regions of India. Indian botanists have persistently used two accepted synonyms of J. adhatoda, namely, Adhatoda vasica and Adhatoda zeylanica, treating them as names of separate species, but without considering or making any reference to variation of forms in the species. Here, different aspects of variation-phenotypic, genotypic, and distributional-in Indian populations of J. adhatoda were studied to determine whether the two names might have been used to designate distinguishable forms of the species. We conducted field studies in different regions of India, laboratory studies of diverse phenotypic traits in experimental plots (anatomical, biochemical, reproductive, and morphometric), and a preliminary study of genetic variation using homologous cytochrome P450 gene fragments. We assessed herbarium samples from across India and the taxonomic literature for pointers indicating the presence of distinguishable forms. Population-level phenotypic and genetic variation pointed to the presence of two distinct morphotypes of the species, which separately tend to occur in dry and wet regions. Each form retains its original phenotype, either when the two forms are transplanted and cultivated together, or when found growing in regions (presumed introduced) outside its normal distributional range. Morphological studies and metabolic profiling (leaf and seed fatty acids, wax load and wax composition in leaf) suggest functional adaptation of the two forms, one to drier and the other to wetter regions. We could distinguish these forms in herbarium specimens dating back to 1821, but neither herbarium specimens nor the taxonomic literature reveal any reference to two forms. We propose that the forms be recognized as two distinct morphotypes of Justicia adhatoda.

Mitochondrial dysfunction and disturbed mitochondrial dynamics were found to be common phenomena in the pathogenesis of Parkinson's disease (PD). Vasicinone is a quinazoline alkaloid from Adhatoda vasica . Here, we investigated the autophagy/mitophagy-enhancing effect of vasicinone and explored its neuroprotective mechanism in paraquat-mimic PD modal in SH-SY5Y cells. Vasicinone rescued the paraquat-induced loss of cell viability and mitochondrial membrane potential. Subsequently, the accumulation of mitochondrial reactive oxygen species (ROS) was balanced by an increase in the expression of antioxidant enzymes. Furthermore, vasicinone restored paraquat-impaired autophagy and mitophagy regulators DJ-1, PINK-1 and Parkin in SH-SY5Y cells. The vasicinone mediated autophagy pathways were abrogated by treatment with the autophagy inhibitor 3-MA, which lead to increases α-synuclein accumulation and decreased the expression of p-ULK and ATG proteins and the autophagy marker LC3-II compared to that observed without 3-MA treatment. These results demonstrated that vasicinone exerted neuroprotective effects by upregulating autophagy and PINK-1/Parkin mediated mitophagy in SH-SY5Y cells., Competing Interests: The authors declare no conflict of interest.

This study aimed to assess and determine the estrogenic activity of the leaf extract of Justicia flava (JF) in mice, which may interfere with its therapeutic use in female reproduction. The uterotrophic assay (UTA) utilizing 20 days old female mice and the reproductive cycle assay (RCA) utilizing adult female mice were used in this study. All administrations were performed orally. Reproductive organ and blood samples were collected the day after last administration of JF for histology and hormone analysis. Other parameters such as organ weight, temperature, body weight, and reproductive cycles were analyzed. Our study showed that for UTA, JF increased uterine weights slightly, which were nonsignificant but more pronounced at the highest dose of 1000 mg/kg. JF did not induce vaginal opening, which is a sign of puberty onset. JF also had minimal effect on organ morphology and caused a slight increase in serum estrogen. For RCA, JF did not significantly alter body weight and temperature although an upward trend in temperature was observed. JF did not disrupt cycling significantly ( P  > .005) compared with estrogen (the positive control drug used). JF also did not significantly alter uterus morphology except at 1000 mg/kg where some increase in the number of glands and cell activity were observed. JF has mild estrogenic activity and will not interfere with reproductive functions at lower doses (10-100 mg/kg) during therapy, but high doses (up to 1000 mg/kg and above) may cause some alterations. Our data, therefore, suggest that JF is a useful candidate in the management of female reproductive health issues at lower doses.

Justicia procumbens is a food and medicine homologous variety, popularly used for making vegetable soups. In this study, a novel mesoporous silica was synthesized and used as the sorbent of SPE for the purification of lignans from J. procumbens. A laboratory-made SPE cartridge was packed with 100 mg of mesoporous silica, which was washed with 10% methanol and eluted using 0.8 mL acetonitrile after sample loading. Afterward, the extract was analyzed by ultra-performance liquid chromatography (UPLC) and MS/MS. All the lignans were efficiently separated in 6 min with the noise level in the range of 50-150 cps. 6'-Hydroxy justicidin B, 6'-hydroxy justicidin A, justicidin B, chinensinaphthol methyl ether, justicidin C, and neojusticdin A were identified to be the dominant molecular species in J. procumbens with contents of 0.065-0.37 mg/g in three tested sample batches from different geographic origins. In conclusion, the proposed mesoporous silica based SPE UPLC-MS/MS method is efficient in linearity (R 2 = 0.9989-0.9996), sensitivity (LOD ≤0.13 μg/kg and LOQ ≤0.42 μg/kg), precision (RSD intra-day ≤3.12 and RSD inter-day ≤4.56), and recovery (83.42-96.11%, RSD ≤2.88%)., (© 2020 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

Indigenous populations use plants as an important healthcare resource or remedy for different diseases. Here, isolated extracts from Justicia (family Acanthanceae) plant leaves used in Africa as remedy for anemia are characterized by different methods to assess composition and potential nutritional or therapeutic value. Extracts from Justicia leaves were obtained by aqueous extraction, with further isolation by centrifuging and high-performance liquid chromatography. Extracts and isolated compounds were characterized by ultraviolet-visible (UV-Vis) spectroscopy and inductively coupled plasma mass spectrometry (ICP-MS). Hemoglobin activity was assessed using different hemoglobin assays (Cayman Chemical, and Sigma-Aldrich), as well as ELISA. In addition, the safety of the isolated samples was assessed in vitro and in vivo in mice. ICP-MS study results revealed many essential metabolites found in blood plasma. The UV-Vis spectroscopy results highlighted the presence of hemoglobin, with assays showing levels over 4 times higher than that of similar mass of lyophilized human hemoglobin. Meanwhile, in vivo studies showed faster recovery from anemia in mice administered with the isolated extracts compared to untreated mice. Moreover, in vitro and in vivo studies highlighted safety of the extracts. This study reveals the presence of high levels of elements essential for blood health in the isolated extracts from Justicia plant leaves. The findings inspire further research with the potential applications in food fortification, and as remedy for blood disorders like anemia, which disproportionally affects cancer patients, pregnant women, and populations in low- and middle-income countries., Competing Interests: The authors declare no conflict of interest. The funders had no role in the design of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript, or in the decision to publish the results.

The noble metal such as Ag and Au doped CeO 2 nanoparticles was successfully prepared by ionic liquid assisted hydrothermal method in the presence of Justicia adhatoda leaves extract and were used as an antibacterial and anticancer agent. The FT-IR and Raman spectrum exhibit the peaks at 460 and 464 cm -1 assigned to CeO stretching vibrations of NPs. The electron microscopic micrographs confirmed the spherical shaped morphology of prepared NPs. The insertion of Ag and Au ions into the CeO 2 surface creates lattice defects the leads to reduce the band gap energy of Ag-Au/CeO 2 at 3.15 eV. The XRD results suggested the average crystalline size of the silver‑gold loaded CeO 2 was 28 nm. From the elemental mapping images, we have visualized that existence and uniform distribution of Ag, Au, Ce, and O in the prepared nanomaterials. The antibacterial activity of unloaded and bimetal loaded CeO 2 NPs was evaluated with Gram-positive and Gram-negative bacteria using disc diffusion assay. The AgAu loaded CeO 2 NPs exhibited the highest zone of inhibition against E. coli and S. aureus strains when compared with pristine CeO 2, Ag loaded CeO 2, and Au loaded CeO 2 NPs. In addition, the 100 μg mL -1 of CeO 2, Ag/CeO 2, Au/CeO 2 and Ag-Au/CeO 2 NPs exposed the 50, 51, 52 and 56% of anticancer activity against the HeLa cells respectively. Overall, this study concludes that the ionic liquid functionalized green synthesized bimetal loaded cerium oxide NPs showed potent antibacterial and anticancer activities., (Copyright © 2019 Elsevier B.V. All rights reserved.)

In folk medicines, Justicia gendarussa ( J. gendarussa ) is used as a depurative herb for treating fever, pain, and cancer and as laxative for constipation. The aim of the present investigation was to evaluate the hepatoprotective effect of the leaf methanol extract of J. gendarussa leaf (J gMe) against carbofuran (CF)-intoxicated liver injuries in Sprague-Dawley rats, along with the antioxidant activity of this extract. For this purpose, levels of serum diagnostic markers, hepatic antioxidant enzymes, and liver histo-architecture were employed to justify the protective efficacy of J gMe. In addition, the phenolic and flavonoid contents of the extract were quantified, and antioxidant activity was investigated using the 1,1-diphenyl-2-picrylhydrazyl (DPPH), nitric oxide, hydrogen peroxide, and hydroxyl free radical scavenging assays. Results revealed that the leaf extract caused a significant (<0.05, <0.01) decrease of the level of hepatic enzymes, triglycerides, and bilirubin and an increase of the total protein. J gMe has also significantly (<0.05, <0.01) lowered the level of malonylaldehyde. Carbofuran markedly suppressed hepatic antioxidant enzymes, however, the leaf extract significantly augmented these enzymes. The hepatoprotective effect was demonstrated by the improvement in the histo-architectural features of liver sections of CF-intoxicated rats treated with J gMe at 500 mg/kg dose. In addition, J gMe showed moderate total phenolic and total flavonoid content, whereas the IC 50 values of DPPH, nitric oxide, hydrogen peroxide, and hydroxyl free radical scavenging assays were 71.31 ± 0.42, 134.82 ± 0.14, 47.69 ± 0.38, and 118.44 ± 0.30 μg/mL, respectively. In conclusion, the present study suggests the protective role of J gMe against hepatic injury induced by CF, which may be attributed to its higher antioxidant properties and thereby scientifically justifies its traditional use.

Twenty-one lignans including three new ones (1, 2 and 13) were isolated from Justicia procumbens. The chemical structures of the new lignans were determined by spectroscopic means including 1D and 2D NMR analysis. These compounds were evaluated for their cytotoxic and anti-HIV activities. The new secoisolariciresinol dimethyl ether acetate (13) exhibited anti-HIV-1 activity with an IC 50 value of 5.27 μmol·L -1 and a selective index (SI) value of 2.2. The known arylnaphthalene lignan procumbenoside A (3) and diphyllin (8) demonstrated inhibitory activity against HIV-1 with IC 50 values of 4.95 (SI > 6.2) and 0.38 μmol·L -1 (SI = 5.3), respectively., (Copyright © 2019 China Pharmaceutical University. Published by Elsevier B.V. All rights reserved.)

Ethnopharmacological Relevance: The leaves of Justicia flava are traditionally used in the South of Nigeria to prevent preterm births., Aim of the Study: In this study, the activity of the methanol leaf extract of J. flava (JF) was investigated on uterine contractility in non-pregnant and pregnant isolated mouse tissues., Material and Methods: The effects on spontaneous, oxytocin, and KCl-induced contractions were determined. The effects in calcium-free media were also determined. Possible mechanisms of activity were investigated using receptor and channel modulators. Mass spectrometric analysis was additionally performed on the leaf extract to identify secondary metabolites., Results: JF was observed to inhibit spontaneous, oxytocin and high KCl-induced uterine contractility. JF also inhibited contractions in Ca 2+ -free media. JF was found to exert its inhibitory effect via interaction with inositol triphosphate and ryanodine receptors and also through modulation of K + - channels. Lignans and alkaloids were identified with the lignans being the most abundant in JF., Conclusion: JF has been shown to potently inhibit uterine contractions in non-pregnant and pregnant isolated mouse uterus. The inhibitory activity of JF has been shown to occur via blockade of extracellular and intracellular calcium entry and these effects may be due to the lignans identified in - JF. JF has therefore been shown in this study to be a lead plant in the discovery of new drugs with uterine inhibitory activity., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Metal and metal oxide nanoparticles (NPs) possess significant properties that are promising materials for biological applications. In this research work, we prepared ionic liquid assisted Ag-Au/ZnO NPs, using J.adhatoda leaves extract by hydrothermal method. Ionic liquids performed as a stabilizing and templating agent to improve the surface morphology of the synthesized Ag and Au doped ZnO NPs. The prepared ZnO, Ag-doped ZnO, Au doped ZnO, and AgAu doped ZnO NPs exhibit the average crystalline size of 36, 34, 32, and 25 nm and their band gap energy values are 3.36, 3.29, 3.17, and 2.98 eV respectively. The XRD and UV-DRS result shows that after doping of Au and Ag the ZnO crystalline size and band gap energy was decreased, which leads to enhanced the biomedical (antibacterial and anticancer) properties of AgAu doped ZnO NPs. The Raman active mode of A 1 (LO) and A 1 (TO) showed that the formation of lattice defects due to the Ag and Au doping in the ZnO crystalline plane to improve the Ag-Au/ZnO properties. SEM and TEM images revealed that the prepared AgAu doped ZnO NPs exhibits nano stick shape with particle size range from 20 to 25 nm. The EDX spectrum and elemental mapping results confirmed that Ag and Au atoms are doped and spread over the ZnO NPs. The corresponding SAED pattern also confirms the crystallinity of Ag-Au/ZnO NPs. Furthermore, the synthesized Ag-Au/ZnO NPs has been explored for its antibacterial and anticancer activities. It shows good antibacterial activity against E.coli and S.aureus bacteria. Additionally, the Ag-Au/ZnO NPs show excellent anticancer activity against the HeLa cancer cells. The excellent antibacterial and anticancer results prove that the bi-metal (Ag and Au) doping can enhance the biomedical properties of ZnO NPs. It will be a promising material in the biomedical field., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Introduction: Polyherbal formulations are an integral part of various indigenous medicinal systems such as Traditional Chinese Medicine (TCM) and Ayurveda. The presence of a very large number of compounds makes the quality control of polyherbal formulations very difficult., Objectives: To overcome this problem, we have developed a comprehensive strategy for the dereplication of natural products in polyherbal formulations by using Adhatoda vasica as a case study., Methods: The strategy is based on five major steps: the collection of plant samples from different locations to observe the effects of environmental variables; LC-ESI-MS/MS-based untargeted metabolite profiling of the plant samples to identify marker compounds using extensive chemometric analysis of the obtained data; the identification of marker compounds in polyherbal products; the isolation, purification and characterization of the marker compounds; and MRM-based quantitative analysis of the isolated marker compounds using LC-ESI-MS/MS., Results: Using this strategy, we identified a total of 51 compounds in the methanolic extract of A. vasica plants from 14 accessions. Chemical fingerprinting of the plant led to the identification of characteristic peaks that were used to confirm the presence of A. vasica in complex polyherbal formulations. Four quinazoline alkaloids (marker compounds) were isolated, purified and quantified in various herbal formulations containing A. vasica., Conclusion: This method demonstrates a comprehensive strategy based on untargeted and targeted metabolite analysis that can be used for the standardization of complex polyherbal formulations.

Objective: To evaluate the effects of Justicia acuminatissima , or Amazonian Sara Tudo , on renal hemodynamics, oxidative profile, and renal histology in rats with ischemic acute kidney injury., Method: Preclinical assay with adult male Wistar rats, weighing from 250 g to 350 g, distributed into Sham, ischemia, and ischemia + Sara Tudo groups. Hemodynamic parameters, renal function, oxidative stress, and renal histology were evaluated., Results: Pretreatment with Sara Tudo reduced the functional injury, which was shown by the increase in creatinine clearance and thiols; reduction of oxidative markers, renal vascular resistance, and tubulointerstitial injury in the renal tissue; and the significant improvement in renal blood flow., Conclusion: The renoprotection provided by Justicia acuminatissima , or Sara Tudo , in cases of ischemic acute kidney injury was characterized by a marked improvement in renal function, reducing the oxidative injury, and impacting on renal histology positively.

The objective of this study was to determine whether preantral follicles cultured in vitro for 7 days within ovine ovarian cortical strips could be isolated at the secondary follicles (SF) and grown until antral stage during an additional 6 days period of in vitro culture in the presence of aqueous extract of Justicia insularis. Fresh ovarian fragments from 16 adult sheep were fixed for histological analysis (Control 1) or in vitro cultured individually in α-MEM + supplemented with 0.3 mg/ml J. insularis (Step 1) for 7 days. Part of the fragments then were fixed for histological analysis (in vitro culture group). Remaining fragments were exposed stepwise to increasing trehalose concentrations before immediate isolation of SF and viability assessment (Control 2) or after 6 days of culture in α-MEM ++ supplemented with 0.3 mg/ml J. insularis (Step 2). In Step 1, percentage of follicular activation was 80%. In Step 2, a significant increase (p < 0.05) in follicular diameter and antrum formation within 6 days in vitro culture of isolated follicles was achieved. The total antioxidant capacity from both steps significantly increase (p < 0.05) from day 2 to day 6. Confocal analysis of oocytes showed 57.14% oocytes with homogeneous distribution and 42.86% with peri-cortical distribution. In conclusion, SF can be successfully isolated from sheep ovarian cortex after 7 days of culture and are capable of surviving and forming an antral cavity if cultured in vitro for an additional 6 days in the presence of 0.3 mg/ml J. insularis., (© 2019 Blackwell Verlag GmbH.)

To analyse metabolic compounds of Justicia adathoda to evaluate against pathogens, inflammation and cervical cancer. The investigation exposed that the extracts of Justicia adathoda have potent metabolic to eradicate the human diseases. The antibacterial, tumorolytic and anti-inflammatory activity of ethyl acetate and aqueous extracts of Justicia adathoda (leaves) were assessed. In vitro anti-inflammatory activity was assessed by standard procedures. Justicia adathoda metabolic exhibit anticancer activity in human cervical cancer cell line (HeLa) (in vitro) analysis. Flavonoids, saponins, alkaloids, amino acids, tannins and terpenoids were present in both the extracts. The active components present in the extracts were found to be amino acids, alkaloids, lipids and triterpenoids which have antibacterial activity shows inhibition against Salmonella and Escherichia coli. Justicia adathoda possesses significant anti-inflammatory activity and it was confirmed by in-vitro analysis. The anticancer activity was found effective in human cervical cancer cell line (HeLa) (in-vitro) analysis. From the investigation could conclude that the metabolic compounds Justicia adathoda is effective against Anti-inflammation and ethyl acetate extract of Justicia adathoda are effective for Cancer.

Background: In Mexico, an infusion made from muicle (Justicia spicigera, Mexican honeysuckle) is used to treat menstrual problems and soothe people's "nerves" (i.e., anxiety and/or depression). However, no studies have yet substantiated these affects. Anxiety currently ranks as the seventh most common cause of disability worldwide, a fact that has intensified the search for new treatments with fewer adverse effects and better therapeutic efficacy., Objective: The anxiolytic-like effect of the aqueous extract of muicle leaves (AEML) was analyzed, and the influence of the estrous cycle on its pharmacological effect in rats was explored., Methods: Female Wistar rats were subjected to the elevated plus maze test to evaluate the anxiolytic effect, followed by an assessment of locomotor activity using the open field test. Diazepam was the reference drug used. Preliminary phytochemical tests were performed., Results: The dose of 12  mg/kg of AEML increased the time to first entry, and the frequency of entries into, the open arms in the proestrus-estrus (P-E) and metestrus-diestrus phases (M-D), while also reducing the anxiety index in the P-E phases. These effects are similar to those observed with diazepam on the elevated plus maze test. None of the treatments showed effects on locomotor activity. The presence of flavonoids, sterols and terpenes in the AEML was confirmed., Conclusion: The anxiolytic-like effect of the dose of 12  mg/kg of AEML in the P-E phases could be due to the presence of the metabolites identified; i.e., flavonoids, sterols and/or terpenes., (Copyright © 2018 Elsevier GmbH. All rights reserved.)

The present study evaluated the effect of supplementing in vitro culture medium with J. insularis compared to FSH on isolated secondary follicles and in vitro maturation of oocytes from those follicles. Secondary follicles were isolated from sheep ovaries and individually cultured for 18 days in α-MEM+ (Control), α-MEM+ supplemented with 100 ng/mL recombinant bovine follicle stimulating hormone (FSH) or with 0.3, 1.25, or 2.5 mg/mL of J. insularis extract (JI0.3, JI1.25, and JI2.5, respectively). Culture medium collected every 2 days was used to measure ROS levels. At the end of the culture period, cumulus oocytes complex (COCs) were collected and matured in vitro. Follicular walls were used for mRNA quantitation. JI0.3 led to a higher (P < 0.05) percentages of intact follicles than other groups after 18 days of culture. While follicular diameter remained unchanged from Day 6 onwards with JI0.3 and FSH, percentages of antral cavity formation were higher (P < 0.05) with JI0.3 at Day 6 than in all other treatments. No differences were observed between controls and treatment groups regarding ROS levels and mRNA expression of genes. Viability of resulting oocytes was higher (P < 0.05) in JI0.3 compared to FSH. Interestingly, in control experiment, supplementation of maturation medium with JI0.3 led to higher (P < 0.05) percentages of metaphase II compared to controls. Although more validations will be needed, it seems that this natural extract could be used as a cheap and easily available alternative to commercial FSH., Competing Interests: The authors have declared that no competing interests exist.

Our previous study revealed that the ethanolic extract of Justicia procumbens ameliorates ovalbumin-induced airway inflammation and airway hyper-responsiveness in a mouse model of asthma. However, the mechanism of action of the extract remains unknown. In this study, we prepared DW2008S, an optimized and standardized powder extracted from J. procumbens using anhydrous ethanol, and investigated its anti-asthmatic effect and mechanism of action. Our results showed that DW2008S contains two major ingredients, justicidin A (JA) and justicidin B (JB), which selectively inhibit T helper 2 (Th2) cell responses in concanavalin A-activated spleen cells and polarized Th2 cells. Blockade of T cell immunoreceptor with immunoglobulin and immunoreceptor tyrosine-based inhibition motif domains (TIGIT) using a neutralizing antibody also selectively inhibited Th2 cell responses. Furthermore, DW2008S regulated TIGIT expression in the mice and cultured cells. Additionally, DW2008S and JA antagonized human adenosine receptor A 3 (A 3 AR), which mediates mast cell-dependent inflammation and bronchoconstriction. DW2008S and JB inhibited human phosphodiesterase 4 (PDE4), which is known to cause bronchoconstriction; however, the required concentrations were higher than those needed to affect TIGIT . These findings suggest that DW2008S can potentially ameliorate Th2-driven airway inflammation and bronchoconstriction through negative regulation of TIGIT and blockade of A 3 AR and PDE4 activities., (© 2018 The Authors. Journal of Cellular and Molecular Medicine published by John Wiley & Sons Ltd and Foundation for Cellular and Molecular Medicine.)