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Research Article | DOI: https://doi.org/10.31579/2688-7517/239
1Department of Medical Laboratory Science, Faculty of Basic Medical Science, College of Medicine, Ekiti State University, Ado-Ekiti, Nigeria.
2Department of Chemistry, Faculty of Science, Federal University, Oye-Ekiti, Nigeria.
3The Department of Science Laboratory Technology, Faculty of Science, Ekiti State University, Ado-Ekiti, Nigeria.
4The Department of Medical Biochemistry, Faculty of Basic Medical Science, College of Medicine, Ekiti State University, Ado-Ekiti, Nigeria.
*Corresponding Author: Adewumi, F. A, Department of Medical Laboratory Science, Faculty of Basic Medical Science, College of Medicine, Ekiti State University, Ado-Ekiti, Nigeria.
Citation: Adewumi, F. A., Oseni, M. O., Asiah, S. E., Oseni, O. A, (2025), Studies on Antimicrobial, Biochemical and Three Metallic Nanoparticles on Aqueous Extracts of Justicia Carnea Leaf, J. Pharmaceutics and Pharmacology Research, 8(3); DOI:10.31579/2688-7517/239
Copyright: © 2025, Adewumi, F. A. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Received: 24 June 2025 | Accepted: 14 July 2025 | Published: 30 July 2025
Keywords: biochemical composition; antioxidant activity; toxicological safety; antimicrobial efficacy; green synthesis
Flamingo Flower (Justicia carnea), historically valued for various applications, and has been researched for its medicinal properties. This study aims to investigate the biochemical composition, antioxidant activity, toxicological safety, antimicrobial efficacy, and green synthesis of the aqueous extract of flamingo flower. Standard analytical methods were used in determining the various parameters. The results of the analyses revealed the presence of alkaloids, flavonoids, tannins, saponins, and phenolic compounds in the extract. It also exhibited strong antioxidant activities against DPPH and NO radicals with increased ferric reducing antioxidant power (FRAP) as well as significant levels of total phenols and flavonoids. The Acute and sub-acute toxicity analyses indicated no adverse effects from the aqueous extract on the Wistar rats. Hematological and white blood cell analyses in animals orally administered with the extract showed a positive safety profile. Furthermore, the green synthesis of metallic nanoparticles (silver, copper, and zinc) which were characterized using UV-Vis spectroscopy and FT-IR showed enhanced antimicrobial activity against various pathogenic bacteria and fungi, revealed some chemical properties based on the amount of light absorbed as well as identified some organic compounds based on their functional groups. Therefore, the study supports the traditional medicinal use of flamingo flower and suggests their potentials for developing natural antimicrobial agents, nanomedicine and drug development.
Justicia carnea, a flowering plant in the Acanthaceae family native to South America, particularly Brazil, is celebrated for its vibrant inflorescences and is widely cultivated worldwide as an ornamental shrub. Typically growing 1-1.5 meters tall with lanceolate to ovate dark green leaves, it produces dense spikes of tubular flowers in shades of pink, red, or white, arranged in verticillasters. These flowers attract pollinators such as bees and butterflies with their nectar-rich blooms. Justicia carnea thrives in well-drained soils with regular watering and partial shade, though it can tolerate full sun in cooler climates [1]. Beyond its aesthetic appeal, recent research has highlighted its potential medicinal properties, including antioxidant and anti-inflammatory activities, which may have implications for pharmaceutical and cosmetic applications. Conservation efforts are crucial due to habitat loss and fragmentation, underscoring the importance of habitat restoration and sustainable harvesting practices. Justicia carnea exemplifies tropical plant diversity, enhancing landscapes and supporting biodiversity through its role as a pollinator attractant [2].
Justicia carnea, a plant renowned for its vibrant flowers, plays a pivotal role in landscaping and horticulture worldwide due to its adaptability to diverse climates and soil conditions [3]. Its ecological significance lies in its ability to attract and support diverse pollinator populations, including bees, butterflies, and hummingbirds, thereby contributing significantly to local biodiversity and ecosystem stability.
Medicinally, Justicia carnea has been studied for its bioactive compounds, such as antioxidants and anti-inflammatory agents, which show promise in combating oxidative stress and inflammation. This potential has sparked interest in pharmacological research aimed at exploring new therapeutic treatments [4]. In terms of cultural importance, Justicia carnea is valued for its visual appeal in gardens and its symbolic representation of beauty and resilience. Its flowers are commonly used in floral arrangements, enhancing landscapes with color and texture. Conservation efforts are increasingly vital to manage Justicia carnea sustainably, preserving its natural habitats and minimizing negative impact on local biodiversity. Continued research and conservation practices are essential to fully understand and harness the benefits of this versatile plant species in various fields [5].
The process of collecting and preparing Justicia carnea (flamingo flower) for aqueous extract involves several critical steps to ensure the extract's integrity and efficacy across various studies. The plant material, specifically the leaves, was collected from Akure, Ondo State, Nigeria, on January 3, 2024, during the early morning to preserve moisture and maximize phytochemical content. The collected specimen underwent authentication and was deposited at the herbarium of the Department of Plant Science, Faculty of Science, Ekiti State University, Ado Ekiti, Nigeria. Authentication was confirmed by Mr. Omotayo, the Chief Technologist, and assigned the code number UHAE 2020068. This meticulous process ensures consistency and reproducibility in biochemical, antioxidant, toxicological, antimicrobial, and nanoparticle synthesis studies involving Justicia carnea [6].
Preparation Of Plant Leaf Extract
The leaves of Justicia Carnea were carefully removed from the plant and rinsed under running tap water to remove any surface dust or contaminants. The cleaned leaves were then air-dried for several days. Once adequately dried, the leaves were ground into a fine powder. A measured amount of 20 grams of this powdered leaf material was mixed with 100 mL of distilled water. This mixture was then agitated continuously overnight to facilitate thorough extraction of the leaf compounds [7].
Aqueous Extraction
Powdered leaves were mixed with distilled water in a 1:10 ratio (w/v) and heated at 70-80°C for 3-4 hours. After cooling to room temperature, the mixture was filtered to separate solid particles from liquid. Concentration was achieved using a rotary evaporator under reduced pressure if needed. The resulting extract was stored at 4°C for stability until future use [8].
Phytochemical Analysis (Fresh Samples)
The phytochemical analysis of fresh plant samples involved tests for flavonoids, alkaloids (Mayer's, Dragendorff's, and Wagner's tests), saponins (foam test), phenolic compounds (Ferric chloride and Lead acetate tests), and steroids (Liebermann's test). Each test utilized specific reagents to detect the presence of these compounds based on characteristic color changes or precipitate formations. Additionally, in vitro antioxidant analysis was conducted using the DPPH free radical scavenging assay [9].
Estimation Of Vitamin C
To estimate the vitamin C content, samples were oxidized with bromine water, neutralized with thiourea, and derivatized with DNPH. After dilution, absorbance at 520 nm was measured against standard ascorbic acid solutions (0.2-1.0 mL, 2 mg/mL). Concentration was calculated using a calibration curve [10].
Green Synthesis and Anti-Microbial Analyses
The experiment involves the biosynthesis of nanoparticles using Justicia carnea leaf extract. Initially, a leaf extract is prepared by mixing sliced leaves with distilled water, filtering, and refrigerating overnight. Three metal ion solutions are then prepared: 0.1 M silver nitrate, 1 mM copper sulfate, and 1 mM zinc sulfate, by dissolving respective salts in deionized water. These solutions serve as precursors for nanoparticle synthesis [11].
Synthesis Of Silver Nanoparticle
Justicia Carnea extract (200 ml) was heated with silver ion solution in a 250 ml flask, leading to a color change indicating nanoparticle formation. The mixture was boiled at 290-700°C for 12 hours, yielding a biphasic solution. The lower nanoparticle-containing layer was centrifuged and rinsed with distilled water to remove unreacted substances. Finally, the nanoparticles were dried in an oven for several days and sent for characterization.
Synthesis Of Copper Nanoparticles
Justicia carnea extract (200 ml) was mixed with a Copper ion solution, causing a color change to dark brown, indicating nanoparticle formation. The mixture was heated for 12 hours, then separated into upper and lower layers in a beaker. The lower layer, containing nanoparticles, was centrifuged and cleaned with distilled water before being oven dried for characterization [12].
Synthesis Of Zinc Nanoparticle
Justicia carnea extract (200 ml) was reacted with zinc ion solution in a 250 ml flask, resulting in a color change to dark brown, indicating reduction of nickel nitrate. The mixture was refluxed for 12 hours at 350°C, then separated into upper and lower layers in a beaker. The lower layer containing nanoparticles was isolated via centrifugation, washed with distilled water, dried, and prepared for characterization [13].
Characterization Of The Synthesized Silver, Copper And Zinc Nanoparticles
Nanoparticle biosynthesis of AgNPs, CuNPs, and ZnNPs was monitored using UV-visible spectroscopy (Shimadzu UV-1800) from 300 to 800 nm. Distilled water served as a blank for baseline correction. FTIR spectroscopy was employed to identify organic functional groups on leaf extract and nanoparticles, revealing their chemical composition and surface characteristics. This dual spectroscopic approach provided comprehensive insights into the synthesis and properties of nanoparticles [14].
Leaves were processed in two sets: one with 5 grams ground and extracted in 50ml ethanol and water, labeled "Aqueous" and "Ethanolic." Another set used 20 grams dry leaves with 300ml ethanol and water for extraction. Extracts were stored refrigerated post-filtration [15].
Liver Marker Enzyme Test
The study evaluated the effects of Justicia carnea extract at doses of 200 mg/kg and 400 mg/kg on liver function in Wistar rats over 21 days. Enzyme levels of ALT, AST, and ALP were measured to assess hepatoprotective or hepatotoxic effects. No acute toxicity was observed, and biochemical analysis showed significant alterations in enzyme levels, indicating potential impacts on liver function at higher doses of the extract. These findings suggest a need for further investigation into the safety and dosage of Justicia carnea extract in therapeutic [16].
Anti- Microbial Analysis
The study prepared concentrations of 100 mg/ml and 50 mg/ml of plant extracts using a double dilution method. Four bacterial isolates—Escherichia coli, Staphylococcus aureus, Salmonella typhi, and Pseudomonas aeruginosa—were tested for antimicrobial activity. Glassware and media were sterilized rigorously, and bacterial inocula were standardized to 0.5 McFarland standard (10^5 cfu/ml). Antimicrobial activity was assessed using Mueller Hinton agar, with the zone of inhibition measured after 24 hours of incubation at 37°C [17].
Table 1: Proximate composition of Flamingo flower (Justicia carnea)
Table 2: Antinutrient composition of Flamingo flower (Justicia carnea)
Table 3: Mineral composition of Flamingo flower (Justicia carnea)
Table 4: Phytochemical screening of Flamingo flower (Justicia carnea)
Table 5: % Antioxidant composition of Flamingo flower (Justicia carnea)
Table 6: Free radical scavenging abilities of Flamingo flower (Justicia carnea)
Table 7b: Antifungal potential of blood leaf (Justicia carnea) aqueous extract and its metal nanoparticles on Fusarium oxysporium
Parameters Mycellial growth inhibition of Fusarium oxysporium (%)
Justicia carnea _
Leaf extract
Justicia carnea 14.82
AgNP
Justicia carnea _
CuNP
Justicia carnea 14.82
ZnNP
|
Measurement Properties]
Wavelength Range (nm.): 200.00 to 900.00
Sampling Interval: 0.5
Scan Mode: Single
Instrument Type: UV-1800 Series
Measuring Mode: Absorbance
Slit Width: 1.0 nm
Light Source Change Wavelength: 340.0 nm
-0.174 200.00 400.00 600.00 800.00 900.00
Figure 1.0a: The UV-Visible Characterization of Blood leaf (Justicia carnea) leaf aqueous extract
| |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
[Measurement Properties]
Wavelength Range(nm.): 200.00 to 900.00
Sampling Interval: 0.5
Scan Mode: Single
Instrument Type:
UV-1800
Series
Measuring Mode: Absorbance
Slit Width: 1.0 nm
Light Source Change Wavelength: 340.0 nm
Figure 1.0d: The UV-Visible Characterization of Blood leaf (Justicia carnea) of zinc nanoparticle
Sample ID: Blood leaf (Justicia carnea) leaf aqueous extract
SampleScans:32 Background Scans:32
Resolution:8 Range:4000 - 400
Figure 2:0a: FTIR characterization of Blood leaf (Justicia carnea) leaf aqueous extract
Sample ID: Blood leaf (Justicia carnea) AgNP
SampleScans:32 Background Scans:32
Resolution:8 Range:4000 - 400
Figure 2:0b: FTIR characterization of Blood leaf (Justicia carnea) Silver nanoparticle (AgNP)
Sample ID: Blood leaf (Justicia carnea) CuNP
SampleScans:32
Background Scans:32
Resolution:8 Range:4000 - 400
Figure 2.0c: FTIR characterization of Blood leaf (Justicia carnea) Copper nanoparticle (CuNP)
Sample ID: Blood leaf (Justicia carnea) ZnNP
SampleScans:32 Background Scans:32
Resolution:8 Range:4000 - 400
Figure 2.0d: FTIR characterization of Blood leaf (Justicia carnea) Zinc nanoparticle (ZnNP)
Justicia carnea (Flamingo flower or Blood leaf) leaves exhibit a diverse nutritional and phytochemical profile [17]. Proximate analysis revealed a moisture content of 10.91 g/100g, ash content of 16.20%, and substantial levels of crude fat (16.67%), crude fibre (15.38%), crude protein (27.71%), and carbohydrates (13.13%). Antinutrient analysis indicated moderate levels of oxalate (0.57 mg/g) and tannins (24.68 mg/L), with low phytate content (0.27%) and significant trypsin inhibitor activity (21.05%).
Mineral composition highlighted significant levels of magnesium, phosphorus, and potassium, with moderate amounts of calcium, iron, and zinc, and trace elements like sodium, copper, chromium, manganese, and lead present. Phytochemical analysis revealed the presence of saponins, phenols, flavonoids (10.0%), terpenoids, steroids, glycosides, alkaloids (5 mg/g), and moderate levels of tannins and phlobatannins [18,19].
Flamingo flower leaves are rich in antioxidants, containing Vitamin C (440.00 mg/100g) and phenolic compounds (1.98 mg GAE/g). Antioxidant assays demonstrated significant free radical scavenging capabilities [20], (e.g., DPPH: 50.43%, NO scavenging: 41.08%) and lipid peroxidation inhibition (TBARS: 0.05 mg MDA/g), along with ferric-reducing antioxidant power (FRAP: 17.39 mg Vitamin C/g) [21].
Further characterization via FTIR spectroscopy of aqueous extracts and metal nanoparticles (AgNP, CuNP, ZnNP) derived from Justicia carnea confirmed the presence of characteristic functional groups such as hydroxyl (-OH), carbonyl (C=O), and methyl/methylene (C-H) groups, indicative of bioactive molecules and stabilizing agents [22]. These findings underscore the nutritional, antioxidant, and potential therapeutic properties of Justicia carnea, supporting its traditional medicinal uses and potential applications in pharmaceutical and dietary contexts [23].
The study revealed that "Ewe Eje," scientifically known as Justicia carnea, is a flowering plant indigenous to parts of tropical Africa, including Nigeria. In traditional medicine, it is valued for its purported therapeutic properties, which include treating skin infections, headaches, and rheumatism. The plant's leaves and roots are typically used, either brewed into teas or applied externally as poultices. The study revealed that Justicia carnea leaf is a good source of phytochemicals which have been reported to have various biochemical and physiological effects. The leaf is apparently a good source of antioxidants and their various nanonoparticles: Zinc nanoparticles, copper nanoparticles and silver nanoparticles have been tested against bacteria and the result showed that they possess antibacterial activities. Vitamin c is present.
The outcome of this research will be useful to all the communities, Pharmacological industries and traditionalist using this plant since they will be the first beneficiaries. It will create awareness to local communities to enhance the traditional knowledge with scientific finding. It will be useful to pharmaceutical companies in order for them to appreciate the benefits, the local users and most communities will be exposed to and the need to gives adequate orientation and advice on planting of the plant.
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To Dear Erin Aust, I would like to express my heartfelt appreciation for the opportunity to have my work published in this esteemed journal. The entire publication process was smooth and well-organized, and I am extremely satisfied with the final result. The Editorial Team demonstrated the utmost professionalism, providing prompt and insightful feedback throughout the review process. Their clear communication and constructive suggestions were invaluable in enhancing my manuscript, and their meticulous attention to detail and dedication to quality are truly commendable. Additionally, the support from the Editorial Office was exceptional. From the initial submission to the final publication, I was guided through every step of the process with great care and professionalism. The team's responsiveness and assistance made the entire experience both easy and stress-free. I am also deeply impressed by the quality and reputation of the journal. It is an honor to have my research featured in such a respected publication, and I am confident that it will make a meaningful contribution to the field.
"I am grateful for the opportunity of contributing to [International Journal of Clinical Case Reports and Reviews] and for the rigorous review process that enhances the quality of research published in your esteemed journal. I sincerely appreciate the time and effort of your team who have dedicatedly helped me in improvising changes and modifying my manuscript. The insightful comments and constructive feedback provided have been invaluable in refining and strengthening my work".
I thank the ‘Journal of Clinical Research and Reports’ for accepting this article for publication. This is a rigorously peer reviewed journal which is on all major global scientific data bases. I note the review process was prompt, thorough and professionally critical. It gave us an insight into a number of important scientific/statistical issues. The review prompted us to review the relevant literature again and look at the limitations of the study. The peer reviewers were open, clear in the instructions and the editorial team was very prompt in their communication. This journal certainly publishes quality research articles. I would recommend the journal for any future publications.
Dear Jessica Magne, with gratitude for the joint work. Fast process of receiving and processing the submitted scientific materials in “Clinical Cardiology and Cardiovascular Interventions”. High level of competence of the editors with clear and correct recommendations and ideas for enriching the article.
We found the peer review process quick and positive in its input. The support from the editorial officer has been very agile, always with the intention of improving the article and taking into account our subsequent corrections.
My article, titled 'No Way Out of the Smartphone Epidemic Without Considering the Insights of Brain Research,' has been republished in the International Journal of Clinical Case Reports and Reviews. The review process was seamless and professional, with the editors being both friendly and supportive. I am deeply grateful for their efforts.
To Dear Erin Aust – Editorial Coordinator of Journal of General Medicine and Clinical Practice! I declare that I am absolutely satisfied with your work carried out with great competence in following the manuscript during the various stages from its receipt, during the revision process to the final acceptance for publication. Thank Prof. Elvira Farina
Dear Jessica, and the super professional team of the ‘Clinical Cardiology and Cardiovascular Interventions’ I am sincerely grateful to the coordinated work of the journal team for the no problem with the submission of my manuscript: “Cardiometabolic Disorders in A Pregnant Woman with Severe Preeclampsia on the Background of Morbid Obesity (Case Report).” The review process by 5 experts was fast, and the comments were professional, which made it more specific and academic, and the process of publication and presentation of the article was excellent. I recommend that my colleagues publish articles in this journal, and I am interested in further scientific cooperation. Sincerely and best wishes, Dr. Oleg Golyanovskiy.
Dear Ashley Rosa, Editorial Coordinator of the journal - Psychology and Mental Health Care. " The process of obtaining publication of my article in the Psychology and Mental Health Journal was positive in all areas. The peer review process resulted in a number of valuable comments, the editorial process was collaborative and timely, and the quality of this journal has been quickly noticed, resulting in alternative journals contacting me to publish with them." Warm regards, Susan Anne Smith, PhD. Australian Breastfeeding Association.
Dear Jessica Magne, Editorial Coordinator, Clinical Cardiology and Cardiovascular Interventions, Auctores Publishing LLC. I appreciate the journal (JCCI) editorial office support, the entire team leads were always ready to help, not only on technical front but also on thorough process. Also, I should thank dear reviewers’ attention to detail and creative approach to teach me and bring new insights by their comments. Surely, more discussions and introduction of other hemodynamic devices would provide better prevention and management of shock states. Your efforts and dedication in presenting educational materials in this journal are commendable. Best wishes from, Farahnaz Fallahian.
Dear Maria Emerson, Editorial Coordinator, International Journal of Clinical Case Reports and Reviews, Auctores Publishing LLC. I am delighted to have published our manuscript, "Acute Colonic Pseudo-Obstruction (ACPO): A rare but serious complication following caesarean section." I want to thank the editorial team, especially Maria Emerson, for their prompt review of the manuscript, quick responses to queries, and overall support. Yours sincerely Dr. Victor Olagundoye.
Dear Ashley Rosa, Editorial Coordinator, International Journal of Clinical Case Reports and Reviews. Many thanks for publishing this manuscript after I lost confidence the editors were most helpful, more than other journals Best wishes from, Susan Anne Smith, PhD. Australian Breastfeeding Association.
Dear Agrippa Hilda, Editorial Coordinator, Journal of Neuroscience and Neurological Surgery. The entire process including article submission, review, revision, and publication was extremely easy. The journal editor was prompt and helpful, and the reviewers contributed to the quality of the paper. Thank you so much! Eric Nussbaum, MD
Dr Hala Al Shaikh This is to acknowledge that the peer review process for the article ’ A Novel Gnrh1 Gene Mutation in Four Omani Male Siblings, Presentation and Management ’ sent to the International Journal of Clinical Case Reports and Reviews was quick and smooth. The editorial office was prompt with easy communication.
Dear Erin Aust, Editorial Coordinator, Journal of General Medicine and Clinical Practice. We are pleased to share our experience with the “Journal of General Medicine and Clinical Practice”, following the successful publication of our article. The peer review process was thorough and constructive, helping to improve the clarity and quality of the manuscript. We are especially thankful to Ms. Erin Aust, the Editorial Coordinator, for her prompt communication and continuous support throughout the process. Her professionalism ensured a smooth and efficient publication experience. The journal upholds high editorial standards, and we highly recommend it to fellow researchers seeking a credible platform for their work. Best wishes By, Dr. Rakhi Mishra.
Dear Jessica Magne, Editorial Coordinator, Clinical Cardiology and Cardiovascular Interventions, Auctores Publishing LLC. The peer review process of the journal of Clinical Cardiology and Cardiovascular Interventions was excellent and fast, as was the support of the editorial office and the quality of the journal. Kind regards Walter F. Riesen Prof. Dr. Dr. h.c. Walter F. Riesen.
Dear Ashley Rosa, Editorial Coordinator, International Journal of Clinical Case Reports and Reviews, Auctores Publishing LLC. Thank you for publishing our article, Exploring Clozapine's Efficacy in Managing Aggression: A Multiple Single-Case Study in Forensic Psychiatry in the international journal of clinical case reports and reviews. We found the peer review process very professional and efficient. The comments were constructive, and the whole process was efficient. On behalf of the co-authors, I would like to thank you for publishing this article. With regards, Dr. Jelle R. Lettinga.
Dear Clarissa Eric, Editorial Coordinator, Journal of Clinical Case Reports and Studies, I would like to express my deep admiration for the exceptional professionalism demonstrated by your journal. I am thoroughly impressed by the speed of the editorial process, the substantive and insightful reviews, and the meticulous preparation of the manuscript for publication. Additionally, I greatly appreciate the courteous and immediate responses from your editorial office to all my inquiries. Best Regards, Dariusz Ziora
Dear Chrystine Mejia, Editorial Coordinator, Journal of Neurodegeneration and Neurorehabilitation, Auctores Publishing LLC, We would like to thank the editorial team for the smooth and high-quality communication leading up to the publication of our article in the Journal of Neurodegeneration and Neurorehabilitation. The reviewers have extensive knowledge in the field, and their relevant questions helped to add value to our publication. Kind regards, Dr. Ravi Shrivastava.
Dear Clarissa Eric, Editorial Coordinator, Journal of Clinical Case Reports and Studies, Auctores Publishing LLC, USA Office: +1-(302)-520-2644. I would like to express my sincere appreciation for the efficient and professional handling of my case report by the ‘Journal of Clinical Case Reports and Studies’. The peer review process was not only fast but also highly constructive—the reviewers’ comments were clear, relevant, and greatly helped me improve the quality and clarity of my manuscript. I also received excellent support from the editorial office throughout the process. Communication was smooth and timely, and I felt well guided at every stage, from submission to publication. The overall quality and rigor of the journal are truly commendable. I am pleased to have published my work with Journal of Clinical Case Reports and Studies, and I look forward to future opportunities for collaboration. Sincerely, Aline Tollet, UCLouvain.
