Publications by authors named "Mohammad K Okla"

27 Publications

  • Page 1 of 1

CuO loaded ZnS nanoflower entrapped on PVA-chitosan matrix for boosted visible light photocatalysis for tetracycline degradation and anti-bacterial application.

J Environ Manage 2022 Jan 11;306:114396. Epub 2022 Jan 11.

Nanobiotechnology Laboratory, Department of Biotechnology, Bannari Amman Institute of Technology, Sathyamangalam, Tamil Nadu, India. Electronic address:

Novel photocatalyst CuO loaded ZnS nanoflower supported on carbon frame work PVA/Chitosan was synthesized by co-precipitation and ultrasonic assisted method. The co-existence of ZnS and CuO and its crystallinity in nanohybrid was verified by XRD, SAED and HR-TEM analysis. The availability of defects in ZnS was identified by EPR. FTIR and TGA verified the presence of PVA and Chitosan. Defects mediated ZnS-CuO/PVA/chitosan heterojunction promote synergistic charge separation with type II interface. Zn-vacancy facilitates two-photon excitation that improves visible-light harvesting. The photocatalytic activity of ZnS-CuO/PVA/Chitosan was 94.7% which is higher when compared to ZnS (40%) and CuO (60%). The photocatalytic mechanism was elucidated using scavenger test and both ·O and ·OH were found to play key role in tetracycline degradation. In addition, ZnS-CuO/PVA/Chitosan demonstrated efficient anti-microbial effect against the both gram strains on comparing with individual ZnS and CuO. Thus, the multifunctional ZnS-CuO/PVA/Chitosan is promising for the photocatalytic degradation of tetracycline and as an antimicrobial agent.
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http://dx.doi.org/10.1016/j.jenvman.2021.114396DOI Listing
January 2022

HPLC-DAD Based Polyphenolic Profiling and Evaluation of Pharmacological Attributes of Wall.

Molecules 2021 Dec 23;27(1). Epub 2021 Dec 23.

Department of Pharmacy, Faculty of Biological Sciences, Quaid-i-Azam University, Islamabad 45320, Pakistan.

The current study was intended to explore the phytochemical profiling and therapeutic activities of Wall. Crude extracts of different plant parts were subjected to the determination of antioxidant, antimicrobial, antidiabetic, cytotoxic, and protein kinase inhibitory potential by using solvents of varying polarity ranges. Maximum phenolic content was notified in distilled water extracts of the stem (DW-S) and leaf (DW-L) while the highest flavonoid content was obtained in ethyl acetate leaf (EA-L) extract. HPLC-DAD analysis confirmed the presence of various polyphenols, quantified in the range of 0.02 ± 0.36 to 2.05 ± 0.18 μg/mg extract. Maximum DPPH scavenging activity was expressed by methanolic extract of the stem (MeOH-S). The highest antioxidant capacity and reducing power was shown by MeOH-S and leaf methanolic extract (MeOH-L), respectively. Proficient antibacterial activity was shown by EA-L extract against and . Remarkable α-amylase and α-glucosidase inhibition potential was expressed by ethyl acetate fruit (EA-F) and n-Hexane leaf (nH-L) extracts, respectively. In case of brine shrimp lethality assay, 41.67% of the extracts (LC < 50 µg/mL) were considered as extremely cytotoxic. The test extracts also showed mild antifungal and protein kinase inhibition activities. The present study explores the therapeutic potential of . and calls for subsequent studies to isolate new bioactive leads through bioactivity-guided isolation.
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http://dx.doi.org/10.3390/molecules27010068DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8746485PMC
December 2021

Impact of Sprouting under Potassium Nitrate Priming on Nitrogen Assimilation and Bioactivity of Three Species.

Plants (Basel) 2021 Dec 27;11(1). Epub 2021 Dec 27.

Integrated Molecular Plant Physiology Research, Department of Biology, University of Antwerp, 2020 Antwerp, Belgium.

Edible sprouts are rich in flavonoids and other polyphenols, as well as proteins, minerals, and vitamins. Increasing sprout consumption necessitates improving their quality, palatability, and bioactivity. The purpose of this study was to test how KNO priming affects the sprouting process species on three species (, , and ) and their nutritional values. Targeted species of were primed with KNO, and the levels of different primary and secondary metabolites were determined. KNO induced biomass accumulation in the sprouts of the three species, accompanied by an increased content of total mineral nutrients, pigments, vitamins, and essential amino acids. Besides, our results showed that KNO enhanced the activity of nitrate reductase (NR), glutamate dehydrogenase (GDH), and glutamine synthetase (GS) enzymes, which are involved in the nitrogen metabolism and GOGAT cycle, which, in turn, increase the nitrogen and protein production. KNO treatment improved the bioactive compound activities of sprouts by increasing total phenolic and flavonoid contents and enhancing the antioxidant and antidiabetic activities. Furthermore, species-specific responses toward KNO priming were noticeable, where showed the highest antioxidant and antidiabetic activities, followed by . Overall, this study sheds the light on the physiological and biochemical bases of growth, metabolism, and tissue quality improvement impact of KNO on sprouts.
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http://dx.doi.org/10.3390/plants11010071DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8747570PMC
December 2021

Synthesis and application of CdS nanoparticles-decorated core-shell [email protected] nanohybrids for visible-light spectrophotometric assay of sulfide in aqueous sample.

Spectrochim Acta A Mol Biomol Spectrosc 2021 Dec 22;270:120793. Epub 2021 Dec 22.

Nanobiotechnology Laboratory, Department of Biotechnology, Bannari Amman Institute of Technology, Sathyamangalam, Tamil Nadu, India. Electronic address:

Novel [email protected] nanosphere decorated with CdS NPs ([email protected] NCs) was synthesized by one step chemical synthesis method. The fabricated NCs were characterized by transmission electron microscope (TEM), scanning electron microscope (SEM), fourier transfer infra-red spectroscopy (FTIR), X-ray diffraction (XRD), X-ray photoelectron spectroscopy (XPS), zeta sizer and particle size analyzer. TEM and XRD confirmed the Ag in core and Ni in shell for the effective formation of [email protected] core shell nanosphere. EDAX and XPS spectra of NCs confirms the formation of [email protected] NCs. Zeta potential and particle size of the NCs was found to be 29.5 ± 1.5 mV and 24 ± 1 nm respectively. The complete loss in the peak intensity of [email protected] NCs (localized surface plasmon resonance (LSPR)) at ∼410 nm in presence of S ions was observed which indicates its selective detection towards S ions. The sulfide ion sensing by [email protected] NCs was due to the successive oxidation of Ag results in the formulation of Ag ions in the system, which causes the diminishing of LSPR band of NCs. The limit of detection (LOD) of S ions by [email protected] NCs was calculated to be of 2.66 nM. The combination of CdS NPs with core-shell [email protected] nanosphere guides a promising strategy for S ions detection from environmental polluted samples.
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http://dx.doi.org/10.1016/j.saa.2021.120793DOI Listing
December 2021

Evaluation of the phycoremediation potential of microalgae for captan removal: Comprehensive analysis on toxicity, detoxification and antioxidants modulation.

J Hazard Mater 2021 Dec 30;427:128177. Epub 2021 Dec 30.

Botany and Microbiology Department, Faculty of Science, Beni-Suef University, Beni-Suef, Egypt; Integrated Molecular Plant Physiology Research, Department of Biology, University of Antwerp, Antwerp, Belgium.

Captan is one of the most widely used organochlorine fungicides, its frequent application contaminates both terrestrial and aquatic ecosystems and negatively affects their key ecological processes. This study demonstrated the toxicity and efficient removal of captan by two different taxonomic species; the green microalga Scenedesmus obliquus and cyanobacterium Nostoc muscorum. After a week of exposure to mild (15 mg/L) and severe (30 mg/L) captan doses, the intracellular captan uptake, degradation and metabolic regulation of captan detoxification were studied. Compared to N. muscorum, S. obliquus accumulated more captan, but efficiently degraded it into two safe eco-friendly by-products; phthalic acid and 1,2,3,6-tetrahydro phthalimide. S. obliquus showed less decrease in cell growth, photosynthesis activity and related parameters including Chla content and activity of PEPC and RuBisCo enzymes. Captan at the severe dose induced oxidative damage particularly in N. muscorum, as expressed by the high levels of HO, MDA, NADPH oxidase and protein peroxidation. Both species invested glutathione-s-transferase enzyme in captan detoxification however, induction of antioxidant defence system e.g. ascorbate and glutathione cycle was more pronounced in S. obliquus which could explain its tolerance ability. This study provided a better understanding of the environmental risks of captan and introduced S. obliquus as a promising captan phycoremediator.
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http://dx.doi.org/10.1016/j.jhazmat.2021.128177DOI Listing
December 2021

Developmental Stages-Specific Response of Anise Plants to Laser-Induced Growth, Nutrients Accumulation, and Essential Oil Metabolism.

Plants (Basel) 2021 Nov 26;10(12). Epub 2021 Nov 26.

Botany and Microbiology Department, Faculty of Science, Beni-Suef University, Beni Suef 62521, Egypt.

Compared to seeds and mature tissues, sprouts are well known for their higher nutritive and biological values. Fruits of (anise) are extensively consumed as food additives; however, the sprouting-induced changes in their nutritious metabolites are hardly studied. Herein, we investigated the bioactive metabolites, phytochemicals, and antioxidant properties of fruits, sprouts (9-day-old), and mature tissue (5-week-old) of anise under laser irradiation treatment (He-Ne laser, 632 nm). Laser treatment increased biomass accumulation of both anise sprouts and mature plants. Bioactive primary (e.g., proteins and sugars) and secondary metabolites (e.g., phenolic compounds), as well as mineral levels, were significantly enhanced by sprouting and/or laser light treatment. Meanwhile, laser light has improved the levels of essential oils and their related precursors (e.g., phenylalanine), as well as enzyme activities [e.g., O-methyltransferase and 3-Deoxy-D-arabino-heptulosonate-7-phosphate synthase (DAHPS)] in mature tissues. Moreover, laser light induced higher levels of antioxidant and anti-lipidemic activities in sprouts as compared to fruits and mature tissues. Particularly at the sprouting stage, anise was more responsive to laser light treatment than mature plants.
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http://dx.doi.org/10.3390/plants10122591DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8708645PMC
November 2021

Anti-Inflammatory Potential of Daturaolone from Mill.: In Silico, In Vitro and In Vivo Studies.

Pharmaceuticals (Basel) 2021 Nov 30;14(12). Epub 2021 Nov 30.

Department of Pharmacy, Quaid-i-Azam University, Islamabad 45320, Pakistan.

Exploration of leads with therapeutic potential in inflammatory disorders is worth pursuing. In line with this, the isolated natural compound daturaolone from Mill. was evaluated for its anti-inflammatory potential using in silico, in vitro and in vivo models. Daturaolone follows Lipinski's drug-likeliness rule with a score of 0.33. Absorption, distribution, metabolism, excretion and toxicity prediction show strong plasma protein binding; gastrointestinal absorption (Caco-2 cells permeability = 34.6 nm/s); no blood-brain barrier penetration; CYP1A2, CYP2C19 and CYP3A4 metabolism; a major metabolic reaction, being aliphatic hydroxylation; no hERG inhibition; and non-carcinogenicity. Predicted molecular targets were mainly inflammatory mediators. Molecular docking depicted H-bonding interaction with nuclear factor kappa beta subunit (NF-κB), cyclooxygenase-2, 5-lipoxygenase, phospholipase A2, serotonin transporter, dopamine receptor D1 and 5-hydroxy tryptamine. Its cytotoxicity (IC) value in normal lymphocytes was >20 µg/mL as compared to cancer cells (Huh7.5; 17.32 ± 1.43 µg/mL). Daturaolone significantly inhibited NF-κB and nitric oxide production with IC values of 1.2 ± 0.8 and 4.51 ± 0.92 µg/mL, respectively. It significantly reduced inflammatory paw edema (81.73 ± 3.16%), heat-induced pain (89.47 ± 9.01% antinociception) and stress-induced depression (68 ± 9.22 s immobility time in tail suspension test). This work suggests a possible anti-inflammatory role of daturaolone; however, detailed mechanistic studies are still necessary to corroborate and extrapolate the findings.
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http://dx.doi.org/10.3390/ph14121248DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8708807PMC
November 2021

Withaferin A: From Ancient Remedy to Potential Drug Candidate.

Molecules 2021 Dec 20;26(24). Epub 2021 Dec 20.

Department of Pharmacy, Faculty of Biological Sciences, Quaid-i-Azam University, Islamabad 45320, Pakistan.

Withaferin A (WA) is a pivotal withanolide that has conquered a conspicuous place in research, owning to its multidimensional biological properties. It is an abundant constituent in Dunal. (Ashwagandha, WS) that is one of the prehistoric pivotal remedies in Ayurveda. This article reviews the literature about the pharmacological profile of WA with special emphasis on its anticancer aspect. We reviewed research publications concerning WA through four databases and provided a descriptive analysis of literature without statistical or qualitative analysis. WA has been found as an effective remedy with multifaceted mechanisms and a broad spectrum of pharmacological profiles. It has anticancer, anti-inflammatory, antiherpetic, antifibrotic, antiplatelet, profibrinolytic, immunosuppressive, antipigmentation, antileishmanial, and healing potentials. Evidence for wide pharmacological actions of WA has been established by both in vivo and in vitro studies. Further, the scientific literature accentuates the role of WA harboring a variable therapeutic spectrum for integrative cancer chemoprevention and cure. WA is a modern drug from traditional medicine that is necessary to be advanced to clinical trials for advocating its utility as a commercial drug.
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http://dx.doi.org/10.3390/molecules26247696DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8705790PMC
December 2021

N.E.Br Manifests Extraction Medium Reliant Disparity in Phytochemical and Pharmacological Analysis.

Molecules 2021 Dec 13;26(24). Epub 2021 Dec 13.

Integrated Molecular Plant Physiology Research, Department of Biology, University of Antwerp, 2020 Antwerpen, Belgium.

Solubility of phytoconstituents depends on the polarity of the extraction medium used, which might result in the different pharmacological responses of extracts. In line with this, ethnomedicinally important food plant (i.e., extracts) have been made in fourteen distinct solvent systems that were then analyzed phytochemically via total phenolic amount estimation, total flavonoid amount estimation, and HPLC detection and quantification of the selected polyphenols. Test extracts were then subjected to a battery of in vitro assays i.e., antioxidants (DDPH scavenging, antioxidant capacity, and reducing power estimation), antimicrobial (antibacterial, antifungal, and antileishmanial), cytotoxic (brine shrimps, THP-1 human leukemia cell lines and normal lymphocytes), and protein kinase inhibition assays. Maximum phenolic and flavonoid contents were computed in distilled water-acetone and acetone extracts (i.e., 16 ± 1 μg/mg extract and 8 ± 0.4/mg extract, respectively). HPLC-DAD quantified rutin (0.58 µg/mg extract) and gallic acid (0.4 µg/mg extract) in methanol-ethyl acetate and methanol extracts, respectively. Water-acetone extract exhibited the highest DPPH scavenging of 36 ± 1%. Total reducing potential of 76.0 ± 1 μg/mg extract was shown by ethanol chloroform while maximum total antioxidant capacity was depicted by the acetone extract (92.21 ± 0.70 μg/mg extract). Maximal antifungal effect against , antileishmanial, brine shrimp cytotoxicity, THP-1 cell line cytotoxicity, and protein kinase inhibitory activities were shown by ethyl acetate-methanol (MIC: 50 µg/disc), n-hexane (IC: 120.8 ± 3.7 µg/mL), ethyl acetate (LD: 29.94 ± 1.6 µg/mL), distilled water-acetone (IC: 118 ± 3.4 µg/mL) and methanol-chloroform (ZOI: 19 ± 1 mm) extracts, respectively. Our findings show the dependency of phytochemicals and bioactivities on the polarity of the extraction solvent and our preliminary screening suggests the extract formulations to be tested and used in different ailments, however, detailed studies remain necessary for corroboration with our results.
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http://dx.doi.org/10.3390/molecules26247530DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8703747PMC
December 2021

Profiling of Antifungal Activities and In Silico Studies of Natural Polyphenols from Some Plants.

Molecules 2021 Nov 26;26(23). Epub 2021 Nov 26.

Department of Biochemistry, Quaid-i-Azam University, Islamabad 45320, Pakistan.

A worldwide increase in the incidence of fungal infections, emergence of new fungal strains, and antifungal resistance to commercially available antibiotics indicate the need to investigate new treatment options for fungal diseases. Therefore, the interest in exploring the antifungal activity of medicinal plants has now been increased to discover phyto-therapeutics in replacement to conventional antifungal drugs. The study was conducted to explore and identify the mechanism of action of antifungal agents of edible plants, including and . The antifungal potential was assessed via the disc diffusion method and, subsequently, the extracts were assessed for phytochemicals and total antioxidant activity. Potent polyphenols were detected using high-performance liquid chromatography (HPLC) and antifungal mechanism of action was evaluated in silico. exhibited antifungal activity against all the tested strains while all plant extracts showed antifungal activity against . Rutin, kaempferol, and quercetin were identified as common polyphenols. In silico studies showed that rutin displayed the greatest affinity with binding pocket of fungal 14-alpha demethylase and nucleoside diphosphokinase with the binding affinity (K, -9.4 and -8.9, respectively), as compared to terbinafine. Results indicated that and exert their antifungal effect possibly due to kaempferol and rutin, respectively, or possibly by inhibition of nucleoside diphosphokinase (NDK) and 14-alpha demethylase (CYP51), while and might exhibit antifungal potential due to quercetin. Overall, the study demonstrates that plant-derived products have a high potential to control fungal infections.
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http://dx.doi.org/10.3390/molecules26237164DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8659076PMC
November 2021

Effect of Elevated CO on Biomolecules' Accumulation in Caraway ( L.) Plants at Different Developmental Stages.

Plants (Basel) 2021 Nov 11;10(11). Epub 2021 Nov 11.

Department of Medicinal and Aromatic Plants, Desert Research Centre, Cairo 11753, Egypt.

Caraway plants have been known as a rich source of phytochemicals, such as flavonoids, monoterpenoid glucosides and alkaloids. In this regard, the application of elevated CO (eCO) as a bio-enhancer for increasing plant growth and phytochemical content has been the focus of many studies; however, the interaction between eCO and plants at different developmental stages has not been extensively explored. Thus, the present study aimed at investigating the changes in growth, photosynthesis and phytochemicals of caraway plants at two developmental stages (sprouts and mature tissues) under control and increased CO conditions (ambient CO (a CO, 400 ± 27 μmol CO mol air) and eCO, 620 ± 42 μmol CO mol air ppm). Moreover, we evaluated the impact of eCO-induced changes in plant metabolites on the antioxidant and antibacterial activities of caraway sprouts and mature plants. CO enrichment increased photosynthesis and biomass accumulation of both caraway stages. Regarding their phytochemical contents, caraway plants interacted differently with eCO, depending on their developmental stages. High levels of CO enhanced the production of total nutrients, i.e., carbohydrates, proteins, fats and crude fibers, as well as organic and amino acids, in an equal pattern in both caraway sprouts and mature plants. Interestingly, the eCO-induced effect on minerals, vitamins and phenolics was more pronounced in caraway sprouts than the mature tissues. Furthermore, the antioxidant and antibacterial activities of caraway plants were enhanced under eCO treatment, particularly at the mature stage. Overall, eCO provoked changes in the phytochemical contents of caraway plants, particularly at the sprouting stage and, hence, improved their nutritive and health-promoting properties.
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http://dx.doi.org/10.3390/plants10112434DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8619137PMC
November 2021

Potential Importance of Molybdenum Priming to Metabolism and Nutritive Value of spp. Sprouts.

Plants (Basel) 2021 Nov 5;10(11). Epub 2021 Nov 5.

Integrated Molecular Plant Physiology Research, Department of Biology, University of Antwerp, 2020 Antwerpen, Belgium.

Molybdenum ions (Mo) can improve plants' nutritional value primarily by enhancing nitrogenous metabolism. In this study, the comparative effects of seed priming using Mo were evaluated among sproutings of species/cultivars, including (CA1), (CA2), and (CA3). Mo impacts on growth, metabolism (e.g., nitrogen and phenolic metabolism, pigment and total nutrient profiles), and biological activities were assayed. Principal component analysis (PCA) was used to correlate Mo-mediated impacts. The results showed that Mo induced photosynthetic pigments that resulted in an improvement in growth and increased biomass. The N content was increased 0.3-fold in CA3 and 0.2-fold in CA1 and CA2. Enhanced nitrogen metabolism by Mo provided the precursors for amino acids, protein, and lipid biosynthesis. At the secondary metabolic level, phenolic metabolism-related precursors and enzyme activities were also differentially increased in species/cultivars. The observed increase in metabolism resulted in the enhancement of the antioxidant (2,2-diphenyl-1-picryl-hydrazyl-hydrate (DPPH) free radical scavenging, 2,2-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), ferric reducing antioxidant power (FRAP)) and antidiabetic potential (Glycemic index (GI) and inhibition activity of α-amylase, and α-glucosidase) of species. The antioxidant activity increased 20% in CA3, 14% in CA1, and 8% in CA2. Furthermore, PCA showed significant variations not only between Mo-treated and untreated samples but also among species. Overall, this study indicated that the sprouts of species have tremendous potential for commercial usage due to their high nutritive value, which can be enhanced further with Mo treatment to accomplish the demand for nutritious feed.
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http://dx.doi.org/10.3390/plants10112387DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8625590PMC
November 2021

Chitosan capped Ag/NiS nanocomposites: A novel colorimetric probe for detection of L-cysteine at nanomolar level and its anti-microbial activity.

Int J Biol Macromol 2021 Dec 10;193(Pt B):2054-2061. Epub 2021 Nov 10.

Nanobiotechnology Laboratory, Department of Biotechnology, Bannari Amman Institute of Technology, Sathyamangalam, Tamil Nadu, India. Electronic address:

L-Cysteine (L-cys) plays very crucial role in biological systems. The study reports the colorimetric detection of L-cys at nanomolar level using chitosan capped Ag decorated NiS nanocomposite (chit-Ag/NiS NCs).The chemical reduction and co-precipitation methods were adopted to prepare chit-Ag/NiS NCs. The fabricated NCs was characterized by X-ray diffraction (XRD), fourier-transform infrared spectroscopy (FT-IR), FT-Raman, scanning electron microscopy (SEM), thermogravimetric analysis (TGA), high-resolution transmission electron microscopy (HR-TEM), energy-dispersive X-ray spectroscopy (EDS) and X-ray photoelectron spectroscopy (XPS). The chit-Ag/NiS NCs particularly detect L-cys even in other amino acids presence. The chit-Ag/NiS NCs showed the surface charge of -26 ± 39.9 mV. The detection of L-cys was indicated by disappearance of yellowish-brown color of Chit-Ag/NiS NCs to colorless. A good linear correlation was found between absorbance vs logarithmic concentration of L-cys (1 μM to 1 nM) with R value of 0.99. The chit-Ag/NiS NCs impregnated cotton swabs was prepared for real time detection of L-cys and the prepared probe was found to be highly selective and specific. The effect of pH, temperature and salinity influencing the L-cys detection was studied. Also, the antimicrobial activity of Chit-Ag/NiS NCs was investigated against gram negative (E. coli) and gram positive (B. subtilis) bacteria.
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http://dx.doi.org/10.1016/j.ijbiomac.2021.11.037DOI Listing
December 2021

Antibacterial, Antihemolytic, Cytotoxic, Anticancer, and Antileishmanial Effects of Transgenic Plants.

Plants (Basel) 2021 Sep 13;10(9). Epub 2021 Sep 13.

Department of Biochemistry, Quaid i Azam University, Islamabad 45320, Pakistan.

Herbal and traditional medicines can play a pivotal role in combating cancer and neglected tropical diseases. , family Lamiaceae, is an important medicinal plant. The genetic transformation of with genes of further enhances its metabolic content. This study aimed at undertaking the molecular, phytochemical, and in vitro biological analysis of extracts. We transformed the plant with genes and raised the regenerants from the hairy roots. Transgenic integration and expression of were confirmed by conventional polymerase chain reaction (PCR) and qPCR analysis. The methanol: chloroform crude extracts of wild-type plants and transgenic regenerants were screened for in vitro antibacterial, antihemolytic, cytotoxic, anticancer, and leishmanial activity. Among all plants, transgenic line 3 (ABRL3) showed the highest expression of the gene. Fourier transform infra-red (FTIR) analysis confirmed the enhanced number of functional groups of active compounds in all transgenic lines. Moreover, ABRL3 exhibited the highest antibacterial activity, minimum hemolytic activity (CC = 7293.05 ± 7 μg/mL) and maximum antileishmanial activity (IC of 56.16 ± 2 μg/mL). ABRL1 demonstrated the most prominent brine shrimp cytotoxicity (LD39.6 ± 4 μg/mL). ABRL3 was most effective against various human cancer cell lines with an IC of 57.1 ± 2.2 μg/mL, 46.2 ± 1.1 μg/mL, 72.4 ± 1.3 μg/mL, 73.3 ± 2.1 μg/mL, 98.7 ± 1.6 μg/mL, and 97.1 ± 2.5 μg/mL against HepG2, LM3, A549, HT29, MCF-7, and MDA-MB-231, respectively. Overall, these transgenic extracts may offer a cheaper therapeutic source than the more expensive synthetic drugs.
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http://dx.doi.org/10.3390/plants10091894DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8473257PMC
September 2021

Polyphenol Rich Transgenic Regenerants Display Better Pharmacological Potential.

Molecules 2021 Aug 11;26(16). Epub 2021 Aug 11.

Department of Biochemistry, Faculty of Biological Sciences, Quaid-i-Azam University, Islamabad 45320, Pakistan.

Wall. ex Benth. is an endangered medicinal herb traditionally used against different ailments. The present study aimed to create new insight into the fundamental mechanisms of genetic transformation and the biological activities of this plant. We transformed the plant with genes of and raised the regenerants from the hairy roots. These transgenic regenerants were screened for in vitro antioxidant activities, a range of in vivo assays, elemental analysis, polyphenol content, and different phytochemicals found through HPLC. Among 18 polyphenolic standards, kaempferol was most abundant in all transgenic lines. Furthermore, transgenic line 3 (ABRL3) showed maximum phenolics and flavonoids content among all tested plant extracts. ABRL3 also demonstrated the highest total antioxidant capacity (8.16 ± 1 μg AAE/mg), total reducing power, (6.60 ± 1.17 μg AAE/mg), DPPH activity (IC50 = 59.5 ± 0.8 μg/mL), hydroxyl ion scavenging (IC50 = 122.5 ± 0.90 μg/mL), and iron-chelating power (IC50 = 154.8 ± 2 μg/mL). Moreover, transformed plant extracts produced significant analgesic, anti-inflammatory, anticoagulant, and antidepressant activities in BALB/c mice models. In conclusion, transgenic regenerants of pose better antioxidant and pharmacological properties under the effect of genes as compared to wild-type plants.
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http://dx.doi.org/10.3390/molecules26164874DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8399588PMC
August 2021

Designing and Development of FRET-Based Nanosensor for Real Time Analysis of N-Acetyl-5-Neuraminic Acid in Living Cells.

Front Nutr 2021 31;8:621273. Epub 2021 May 31.

Department of Botany, Faculty of Life Sciences, Aligarh Muslim University, Aligarh, India.

-acetyl-5-neuraminic acid (NeuAc) plays crucial role in improving the growth, brain development, brain health maintenance, and immunity enhancement of infants. Commercially, it is used in the production of antiviral drugs, infant milk formulas, cosmetics, dietary supplements, and pharmaceutical products. Because of the rapidly increasing demand, metabolic engineering approach has attracted increasing attention for NeuAc biosynthesis. However, knowledge of metabolite flux in biosynthetic pathways is one of the major challenges in the practice of metabolic engineering. So, an understanding of the flux of NeuAc is needed to determine its cellular level at real time. The analysis of the flux can only be performed using a tool that has the capacity to measure metabolite level in cells without affecting other metabolic processes. A Fluorescence Resonance Energy Transfer (FRET)-based genetically-encoded nanosensor has been generated in this study to monitor the level of NeuAc in prokaryotic and eukaryotic cells. Sialic acid periplasmic binding protein (SiaP) from was exploited as a sensory element for the generation of nanosensor. The enhanced cyan fluorescent protein (ECFP) and Venus were used as Fluroscence Resonance Energy Transfer (FRET) pair. The nanosensor, which was termed fluorescent indicator protein for sialic acid (FLIP-SA), was successfully transformed into, and expressed in BL21 (DE3) cells. The expressed protein of the nanosensor was isolated and purified. The purified nanosensor protein was characterized to assess the affinity, specificity, and stability in the pH range. The developed nanosensor exhibited FRET change after addition to NeuAc. The developed nanosensor was highly specific, exhibited pH stability, and detected NeuAc levels in the nanomolar to milimolar range. FLIP-SA was successfully introduced in bacterial and yeast cells and reported the real-time intracellular levels of NeuAc non-invasively. The FLIP-SA is an excellent tool for the metabolic flux analysis of the NeuAc biosynthetic pathway and, thus, may help unravel the regulatory mechanism of the metabolic pathway of NeuAc. Furthermore, FLIP-SA can be used for the high-throughput screening of mutant libraries for varied NeuAc production levels.
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http://dx.doi.org/10.3389/fnut.2021.621273DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8200523PMC
May 2021

A novel biogenic Allium cepa leaf mediated silver nanoparticles for antimicrobial, antioxidant, and anticancer effects on MCF-7 cell line.

Environ Res 2021 07 29;198:111199. Epub 2021 Apr 29.

Department of Botany and Microbiology, College of Science, King Saud University, Riyadh, 11451, Saudi Arabia.

In the present study, Allium cepa leaf extract was utilized to reduce the silver nitrate into the nanoscale range of silver ions (Ag NPs). The biosynthesized Ag NPs were extensively characterized by X-ray diffraction analysis (XRD), Dynamic light scattering analysis (DLS), UV-Visible spectroscopy (UV-vis), Transmission electron microscopy (TEM), Energy dispersive X-ray analysis (EDX) and Fourier transform infrared spectroscopy (FTIR). The antioxidant activity of synthesized Ag NPs was verified by DPPH assay. From the results obtained from XRD and DLS studies, the size of Ag NPs was determined to be around 54.3 nm. The measured zeta potential value of -19.1 mV confirms the excellent stability of biosynthesized Ag NPs. TEM analyses reveal that the biosynthesized Ag NPs have a spherical structure of 13 nm in size. The presence of various functional groups was confirmed through FTIR studies and EDAX verifies the weight percentage of silver content in biosynthesized nanoparticles to be 30.33%. In the present study, anti-cancer activity was carried out by using breast cancer cell line MCF-7. Further, silver nanoparticles exhibited antimicrobial effectiveness against gram-positive Bacillus cereus and gram-negative Escherichia coli. The MTT assay also showed better cytotoxic activity against the MCF- 7 cell line.
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http://dx.doi.org/10.1016/j.envres.2021.111199DOI Listing
July 2021

FRET-Based Genetically Encoded Nanosensor for Real-Time Monitoring of the Flux of α-Tocopherol in Living Cells.

ACS Omega 2021 Apr 23;6(13):9020-9027. Epub 2021 Mar 23.

Department of Botany, Aligarh Muslim University, Aligarh 202002, India.

Vitamin E plays an exemplary role in living organisms. α-Tocopherol is the most superior and active form of naturally occurring vitamin E that meets the requirements of human beings as it possesses the α-tocopherol transfer protein (α-TTP). α-Tocopherol deficiency can lead to severe anemia, certain cancers, several neurodegenerative and cardiovascular diseases, and most importantly male infertility. As a result of the depletion of its natural sources, researchers have tried to employ metabolic engineering to enhance α-tocopherol production to meet the human consumption demand. However, the metabolic engineering approach relies on the metabolic flux of a metabolite in its biosynthetic pathway. Analysis of the metabolic flux of a metabolite needs a method that can monitor the α-tocopherol level in living cells. This study was undertaken to construct a FRET (fluorescence resonance energy transfer)-based nanosensor for monitoring the α-tocopherol flux in prokaryotic and eukaryotic living cells. The human α-TTP was sandwiched between a pair of FRET fluorophores to construct the nanosensor, which was denoted as FLIP-α (the fluorescence indicator for α-tocopherol). FLIP-α showed excellence in monitoring the α-tocopherol flux with high specificity. The sensor was examined for its pH stability for physiological applications, where it shows no pH hindrance to its activity. The calculated affinity of this nanosensor was 100 μM. It monitored the real-time flux of α-tocopherol in bacterial and yeast cells, proving its biocompatibility in monitoring the α-tocopherol dynamics in living cells. Being noninvasive, FLIP-α provides high temporal and spatial resolutions, which holds an indispensable significance in bioimaging metabolic pathways that are highly compartmentalized.
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http://dx.doi.org/10.1021/acsomega.1c00041DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8028167PMC
April 2021

Antibacterial and Antifungal Activity of the Extracts of Different Parts of (Forssk.) Vierh.

Plants (Basel) 2021 Jan 28;10(2). Epub 2021 Jan 28.

Department of Zoology, Faculty of Science, King Saud University, Riyadh 11451, Saudi Arabia.

Increased problems associated with side effects and bacterial resistance of chemical drugs has prompted the research focus on herbal medicines in the past few decades. In the present investigation, the antimicrobial activity of the various parts of (AM), a mangrove plant, has been evaluated. The plants were collected from the Jazan area of the Kingdom of Saudi Arabia. Primary extracts of roots, stem, leaves, fruits, and seeds were made in ethanol and fractioned in ethanol, ethyl acetate, petroleum ether, chloroform, and water. Minimal inhibitory concentration (MIC) and minimal bactericidal concentration (MBC) of the extracts were determined against , , , and . It has been observed that the chloroform extract of roots of the AM exhibited inhibitory effects against both (MIC = 1.5 ± 0.03 mg/mL) and (MIC = 1.7 ± 0.01 mg/mL). The ethanolic extract of the AM roots has shown antibacterial activity against (MIC = 10.8 ± 0.78 mg/mL) (MIC = 6.1 ± 0.27 mg/mL), (MIC = 2.3 ± 0.08 mg/mL), and (MIC = 6.3 ± 0.28 mg/mL). The leaf extract of the AM in ethyl acetate showed antibacterial activity against and . Antifungal activity of these extracts was also investigated against and Ethanolic extract of roots and seeds of the AM has shown antifungal activity against when applied individually. Ethanolic extract of the AM fruits has shown an inhibitory effect on the growth of and . It is suggested that the plant extracts of AM have tremendous antimicrobial activity against a group of microbes, and this effect depends on both the plant part and the solvent used for extraction. Therefore, this plant can be considered to treat various diseases caused by antibiotic-resistant bacteria.
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http://dx.doi.org/10.3390/plants10020252DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7911470PMC
January 2021

A Non-Invasive Tool for Real-Time Measurement of Sulfate in Living Cells.

Int J Mol Sci 2020 Apr 7;21(7). Epub 2020 Apr 7.

Department of Botany, Faculty of Life Sciences, Aligarh Muslim University, Aligarh 202001, India.

Sulfur (S) is an essential element for all forms of life. It is involved in numerous essential processes because S is considered as the primary source of one of the essential amino acids, methionine, which plays an important role in biological events. For the control and regulation of sulfate in a metabolic network through fluxomics, a non-invasive tool is highly desirable that opens the door to monitor the level of the sulfate in real time and space in living cells without fractionation of the cells or tissue. Here, we engineered a FRET (fluorescence resonance energy transfer) based sensor for sulfate, which is genetically-encoded and named as FLIP-SP (Fluorescent indicator protein for sulfate). The FLIP-SP can measure the level of the sulfate in live cells. This sensor was constructed by the fusion of fluorescent proteins at the N- and C-terminus of sulfate binding protein (sbp). The FLIP-SP is highly specific to sulfate, and showed pH stability. Real-time monitoring of the level of sulfate in prokaryotic and eukaryotic cells showed sensor bio-compatibility with living cells. We expect that this sulfate sensor offers a valuable strategy in the understanding of the regulation of the flux of sulfate in the metabolic network.
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http://dx.doi.org/10.3390/ijms21072572DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7177696PMC
April 2020

Metabolic Flux Analysis of Catechin Biosynthesis Pathways Using Nanosensor.

Antioxidants (Basel) 2020 Mar 31;9(4). Epub 2020 Mar 31.

Department of Botany, Aligarh Muslim University, Aligarh 202002, India.

(+)-Catechin is an important antioxidant of green tea ( (L.) O. Kuntze). Catechin is known for its positive role in anticancerous activity, extracellular matrix degradation, cell death regulation, diabetes, and other related disorders. As a result of enormous interest in and great demand for catechin, its biosynthesis using metabolic engineering has become the subject of concentrated research with the aim of enhancing (+)-catechin production. Metabolic flux is an essential concept in the practice of metabolic engineering as it helps in the identification of the regulatory element of a biosynthetic pathway. In the present study, an attempt was made to analyze the metabolic flux of the (+)-catechin biosynthesis pathway in order to decipher the regulatory element of this pathway. Firstly, a genetically encoded fluorescence resonance energy transfer (FRET)-based nanosensor (FLIP-Cat, fluorescence indicator protein for (+)-catechin) was developed for real-time monitoring of (+)-catechin flux. characterization of the purified protein of the nanosensor showed that the nanosensor was pH stable and (+)-catechin specific. Its calculated was 139 µM. The nanosensor also performed real-time monitoring of (+)-catechin in bacterial cells. In the second step of this study, an entire (+)-catechin biosynthesis pathway was constructed and expressed in in two sets of plasmid constructs: pET26b-PT7-rbs-PAL-PT7-rbs-4CL-PT7-rbs-CHS-PT7-rbs-CHI and pET26b-T7-rbs-F3H-PT7-rbs- DFR-PT7-rbs-LCR. The harboring the FLIP-Cat was transformed with these plasmid constructs. The metabolic flux analysis of (+)-catechin was carried out using the FLIP-Cat. The FLIP-Cat successfully monitored the flux of catechin after adding tyrosine, 4-coumaric acid, 4-coumaroyl CoA, naringenin chalcone, naringenin, dihydroquercetin, and leucocyanidin, individually, with the bacterial cells expressing the nanosensor as well as the genes of the (+)-catechin biosynthesis pathway. Dihydroflavonol reductase (DFR) was identified as the main regulatory element of the (+)-catechin biosynthesis pathway. Information about this regulatory element of the (+)-catechin biosynthesis pathway can be used for manipulating the (+)-catechin biosynthesis pathway using a metabolic engineering approach to enhance production of (+)-catechin.
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http://dx.doi.org/10.3390/antiox9040288DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7222200PMC
March 2020

Development of an In Vitro Propagation Protocol and a Sequence Characterized Amplified Region (SCAR) Marker of Wall. ex Ging.

Plants (Basel) 2020 Feb 14;9(2). Epub 2020 Feb 14.

Department of Botany, Aligarh Muslim University, Aligarh 202002, India.

An efficient protocol of plant regeneration through indirect organogenesis in was developed in the present study. Culture of leaf explants on MS (Murashige and Skoog) medium supplemented with 2.0 mg/L 6-benzyladenine and 0.13 mg/L 2,4-dichloro phenoxy acetic acid. Adventitious shoot formation was observed when calli were transferred on to MS medium containing 0.5 mg/L α-naphthalene acetic acid and 2.25 mg/L kinetin, which showed the maximum 86% shoot regeneration frequency. The highest root frequency (80.92%) with the 5.6 roots per explant and 1.87 cm root length was observed on MS medium supplemented with 2 mg/L indole-3-butyric acid. The plantlets were transferred to the mixture of sand, coffee husk and soil in the ratio of 1:2:1 in a pot, and placed under 80% shade net for one month. It was then transferred to 30% shade net for another one month, prior to transplantation in the field. These plantlets successfully acclimatized under field conditions. A Sequence Characterized Amplified Region (SCAR) marker was also developed using a 1135 bp amplicon that was obtained from RAPD (Random Amplification of Polymorphic DNA) analysis of six accessions of Testing of several market samples of using the SCAR marker revealed successful identification of the genuine samples of This study, therefore, provides a proficient in vitro propagation protocol of using leaf explants and a SCAR marker for the authentic identification of . This study will be helpful for conservation of authentic .
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http://dx.doi.org/10.3390/plants9020246DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7076368PMC
February 2020

Novel halochromic cellulose nanowhiskers from rice straw: Visual detection of urea.

Carbohydr Polym 2020 Mar 13;231:115740. Epub 2019 Dec 13.

Department of Zoology, Faculty of Science, Beni-Suef University, Beni-Suef, 65211, Egypt.

Colorimetric nanocomposite film sensor was fabricated by incorporating TCFH spectroscopic probe into cellulose nanowhiskers (CNW)/Urease enzyme matrix. CNW-TCFH can be used as disposable molecular biosensor in which CNW is the probe carrier comprising high surface area-to-volume ratio, urease is the catalyst and TCFH is the molecular probe. Tricyanofuran-hydrazone (TCFH) spectroscopic probe was prepared. UV-vis absorption spectra demonstrated solvatochromic behavior and a reversible color change of the tricyanofuran-hydrazone probe solution in acetone under acid/base conditions. CNW were reinforced with sodium alginate biopolymer to introduce biocomposite film. This CNW-TCFH film biosensor responds through visible color shift from light yellow to pink when exposed to urea in aqueous media. The morphology properties of CNW and CNW-TCFH films were examined by different tools. The photophysical properties of the prepared TCFH probe, including solvatochromism and pH sensory, were also studied.
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http://dx.doi.org/10.1016/j.carbpol.2019.115740DOI Listing
March 2020

Antioxidant, Hypoglycemic, and Neurobehavioral Effects of a Leaf Extract of on Autoimmune Diabetic Mice.

Evid Based Complement Alternat Med 2019 21;2019:1263260. Epub 2019 May 21.

Department of Zoology, Faculty of Science, King Saud University, Riyadh, Saudi Arabia.

Diabetes mellitus (DM) is a metabolic disease that can affect the central nervous system and behavioral traits in animals. Streptozotocin-induced diabetes is considered an autoimmune disease. The aim of the current study was to determine whether supplementation with the alcoholic extract of leaves could improve diabetes-associated pathological changes. The animals were divided into four groups: a control group (A), an receiving nondiabetic group (B), a diabetic group (C), and a DM group orally supplemented with alcoholic leaf extract (D). The DM group of animals receiving the alcoholic extract of leaves had reduced blood glucose levels, improved blood picture, and organ functions. This group also showed improvement in locomotory behavior. The results of this study showed that supplementation with the alcoholic extract of leaves reduced oxidative stress and blood sugar levels, protected the liver, and improved the neurobehavioral changes associated with diabetes in mice. Introducing alcoholic leaf extract of to diabetic mice decreased inflammatory cells aggregation, vacuolation, and hemorrhage. Additionally, a positive effect of the alcoholic leaf extract on the histopathological changes was observed in the testicular tissue of treated mice.
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http://dx.doi.org/10.1155/2019/1263260DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6556331PMC
May 2019

Molecular and phytochemical analysis of wild type and olive cultivars grown under Saudi Arabian environment.

3 Biotech 2017 Oct 24;7(5):289. Epub 2017 Aug 24.

Department of Plant Production College of Food and Agricultural Sciences, King Saud University, Riyadh, Saudi Arabia.

This study aimed to assess genetic variability at molecular and phytochemical levels among the four most commonly grown olive cultivars and the wild-type olive of Saudi Arabia. Sixty-six and 80 amplicons were generated from 9 random amplified polymorphic DNA (RAPD) and inter simple sequence repeats (ISSR) primers, each, producing an average of 95.9 and 86.44% polymorphism for the two markers, respectively. The PIC values were 82.2% for the RAPD and 85.4% for the ISSR markers and the discrimination power for both the markers was 11.1%. The UPGMA cluster analysis based on the RAPD and ISSR data resulted in the aggregation of cultivars and wild accession with a good bootstrapping value according to their origin. Furthermore, a total of 199 compounds were identified in the cultivars based on peak area, retention time, and molecular formula using GC-MS analyses of methanolic and ethanolic extracts. These compounds were classified according to their chemical class; most of them were fatty acids, alcoholic compounds, carboxylic acids, aldehydes, heterocyclic compounds, ketones, alkanes, and phenols. Genetic and phytochemical distances were significantly correlated, based on the Mantel test. The Saudi wild accession also had high numbers of fatty acids and their esters, and can be used in breeding programs for generating new genotypes with interesting characters.
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http://dx.doi.org/10.1007/s13205-017-0920-5DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5570721PMC
October 2017

Evaluation of Skin Surface as an Alternative Source of Reference DNA Samples: A Pilot Study.

J Forensic Sci 2018 Jan 23;63(1):227-233. Epub 2017 Feb 23.

College of Forensic Sciences, Naif Arab University for Security Sciences, P.O. Box: 6830, Riyadh, 11452, Saudi Arabia.

An acceptable area for collecting DNA reference sample is a part of the forensic DNA analysis development. The aim of this study was to evaluate skin surface cells (SSC) as an alternate source of reference DNA sample. From each volunteer (n = 10), six samples from skin surface areas (forearm and fingertips) and two traditional samples (blood and buccal cells) were collected. Genomic DNA was extracted and quantified then genotyped using standard techniques. The highest DNA concentration of SSC samples was collected using the tape/forearm method of collection (2.1 ng/μL). Cotton swabs moistened with ethanol yielded higher quantities of DNA than swabs moistened with salicylic acid, and it gave the highest percentage of full STR profiles (97%). This study supports the use of SSC as a noninvasive sampling technique and as a extremely useful source of DNA reference samples among certain cultures where the use of buccal swabs can be considered socially unacceptable.
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http://dx.doi.org/10.1111/1556-4029.13468DOI Listing
January 2018

Genotoxicity of ferric oxide nanoparticles in Raphanus sativus: Deciphering the role of signaling factors, oxidative stress and cell death.

J Environ Sci (China) 2016 Sep 18;47:49-62. Epub 2016 Apr 18.

Zoology Department, College of Sciences, King Saud University, P.O. Box 2455, Riyadh 11451, Saudi Arabia; A.R. Al-Jeraisy Chair for DNA Research, Zoology Department, College of Sciences, King Saud University, P.O. Box 2455, Riyadh 11451, Saudi Arabia.

We have studied the genotoxic and apoptotic potential of ferric oxide nanoparticles (Fe2O3-NPs) in Raphanus sativus (radish). Fe2O3-NPs retarded the root length and seed germination in radish. Ultrathin sections of treated roots showed subcellular localization of Fe2O3-NPs, along with the appearance of damaged mitochondria and excessive vacuolization. Flow cytometric analysis of Fe2O3-NPs (1.0mg/mL) treated groups exhibited 219.5%, 161%, 120.4% and 161.4% increase in intracellular reactive oxygen species (ROS), mitochondrial membrane potential (ΔΨm), nitric oxide (NO) and Ca(2+) influx in radish protoplasts. A concentration dependent increase in the antioxidative enzymes glutathione (GSH), catalase (CAT), superoxide dismutase (SOD) and lipid peroxidation (LPO) has been recorded. Comet assay showed a concentration dependent increase in deoxyribonucleic acid (DNA) strand breaks in Fe2O3-NPs treated groups. Cell cycle analysis revealed 88.4% of cells in sub-G1 apoptotic phase, suggesting cell death in Fe2O3-NPs (2.0mg/mL) treated group. Taking together, the genotoxicity induced by Fe2O3-NPs highlights the importance of environmental risk associated with improper disposal of nanoparticles (NPs) and radish can serve as a good indicator for measuring the phytotoxicity of NPs grown in NP-polluted environment.
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http://dx.doi.org/10.1016/j.jes.2015.12.037DOI Listing
September 2016
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