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Ziziphus oxyphylla Edgew is in folk use in Pakistan as an analgesic, anti-inflammatory, and liver ailments. Therefore, we have investigated antioxidant, antinociceptive, anti-inflammatory, and hepatoprotective activities of the isolated compounds (ceanothic acid and zizybrenalic acid) from the chloroform fraction of Z. oxyphylla . Ceanothic acid and zizybrenalic acid showed significant DPPH and H 2 O 2 scavenging activity as compared to control. In the acute toxicity study, ceanothic acid and zizybrenalic acid showed no toxic effects upto 200 mg/kg. The antinociceptive activity shown by ceanothic acid and zizybrenalic acid at 50 mg/kg was 64.28% and 65.35% compared to diclofenac sodium (72.3%) at 50 mg/kg. The percent inhibition of xylene-induced ear edema exhibited by ceanothic acid and zizybrenalic acid at 50 mg/kg was 51.33% and 58.66%, respectively, as compared to diclofenac sodium (72.66%). Both the isolated compounds exhibited inhibition of carrageenan-induced paw edema as compared to control. Hepatoprotection exhibited by zizybrenalic acid was more pronounced than ceanothic acid as observed from the decrease in carbon tetrachloride (CCl 4 )-induced elevation of serum biomarkers, antioxidant enzymes and lipid peroxidation. Furthermore, zizybrenalic acid produced a marked decline in CCl 4 -induced prolongation of phenobarbital-induced sleeping duration. Zizybrenalic acid exhibited 55.4 ± 1.37% inhibition of hypotonic solution-induced hemolysis compared to sodium salicylate (75.6 ± 2.15%). The histopathological damage caused by CCl 4 was also countered by the administration of ceanothic acid and zizybrenalic acid. Ceanothic acid and zizybrenalic acid exhibited antioxidant, antinociceptive, anti-inflammatory, and hepatoprotective activities. Zizybrenalic acid exhibited better antioxidant, antinociceptive, anti-inflammatory, and hepatoprotective activity than ceanothic acid.

Fagonia indica is a rich source of pharmacologically active compounds. The variation in the metabolites of interest is one of the major issues in wild plants due to different environmental factors. The addition of chemical elicitors is one of the effective strategies to trigger the biosynthetic pathways for the release of a higher quantity of bioactive compounds. Therefore, this study was designed to investigate the effects of chemical elicitors, aluminum chloride (AlCl 3 ) and cadmium chloride (CdCl 2 ), on the biosynthesis of secondary metabolites, biomass, and the antioxidant system in callus cultures of F. indica . Among various treatments applied, AlCl 3 (0.1 mM concentration) improved the highest in biomass accumulation (fresh weight (FW): 404.72 g/L) as compared to the control (FW: 269.85 g/L). The exposure of cultures to AlCl 3 (0.01 mM) enhanced the accumulation of secondary metabolites, and the total phenolic contents (TPCs: 7.74 mg/g DW) and total flavonoid contents (TFCs: 1.07 mg/g DW) were higher than those of cultures exposed to CdCl 2 (0.01 mM) with content levels (TPC: 5.60 and TFC: 0.97 mg/g) as compared to the control (TPC: 4.16 and TFC: 0.42 mg/g DW). Likewise, AlCl 3 and CdCl 2 also promoted the free radical scavenging activity (FRSA; 89.4% and 90%, respectively) at a concentration of 0.01 mM, as compared to the control (65.48%). For instance, the quantification of metabolites via high-performance liquid chromatography (HPLC) revealed an optimum production of myricetin (1.20 mg/g), apigenin (0.83 mg/g), isorhamnetin (0.70 mg/g), and kaempferol (0.64 mg/g). Cultures grown in the presence of AlCl 3 triggered higher quantities of secondary metabolites than those grown in the presence of CdCl 2 (0.79, 0.74, 0.57, and 0.67 mg/g). Moreover, AlCl 3 at 0.1 mM enhanced the biosynthesis of superoxide dismutase (SOD: 0.08 nM/min/mg-FW) and peroxidase enzymes (POD: 2.37 nM/min/mg-FW), while CdCl 2 resulted in an SOD activity up to 0.06 nM/min/mg-FW and POD: 2.72 nM/min/mg-FW. From these results, it is clear that AlCl 3 is a better elicitor in terms of a higher and uniform productivity of biomass, secondary cell products, and antioxidant enzymes compared to CdCl 2 and the control. It is possible to scale the current strategy to a bioreactor for a higher productivity of metabolites of interest for various pharmaceutical industries.

Background: Viola canescens Wall. ex. Roxb. exhibits analgesic, antimalarial and antispasmodic activities. It is used folklorically for the treatment of liver diseases, hypertension, malaria and cancer. The current study investigates phytochemical constituents, antioxidant and hepatoprotective activity of solvent extracts of whole plant of Viola canescens., Methods: Phytochemicals, acute toxicity study and antioxidant activity of Viola canescens methanolic extract (VCME), ethyl acetate fraction (EAF), and partially purified EAF (90% EAF and combination of 80% EAF + 20% methanol fraction (EAF + Me) was carried out. Hepatoprotective activity of VCME, EAF (200 and 400 mg/kg body weight) and partially purified EAF (50 mg/kg body weight) was investigated in carbon tetrachloride (CCl 4 ) intoxicated BALB/c mice for 7 days. Membrane stabilization effect was determined by hypotonic solution induced hemolysis while DNA ladder assay was carried out by polyacrylamide gel electrophoresis., Results: Phytochemical screening of VCME showed the presence of alkaloids, phenols, flavonoids, saponins, carbohydrates, tannins and triterpenes. VCME, EAF (at 200 and 400 mg/kg body weight) and partially purified EAF (90% EAF and EAF + Me) at 50 mg/kg body weight significantly reduced the level of ALT, ALP, total bilirubin and restored the level of serum protein in comparison to CCl 4 treated group. A significant reduction in malondialdehyde (MDA) and elevation in catalase (CAT) and superoxide dismutase (SOD) level was observed in extract treated animals as compared to CCl 4 (p < 0.05). The IC 50 values in membrane stabilization potential for VCME, EAF and sodium salicylate were 3.7 ± 0.11, 3.4 ± 0.15 and 3.2 ± 0.09 mg/ml, respectively. Similarly, CCl 4 induced degradation of DNA was counteracted by VCME and EAF. The liver biopsy of mice treated with the solvent extracts showed remarkable restoration of normal histological archeitecture., Conclusions: Viola canescens showed significant hepatoprotective potential due to its antioxidant and membrane stabilization effect.

Gentamicin is a potent antibiotic, effective against Gram negative bacteria. The most common adverse effect of gentamicin is nephrotoxicity. Present study was aimed to explore the protective potentials of Citrullus colocynthis against gentamicin induced nephrotoxicity due to its strong antioxidant properties. Toxic doses of gentamicin (80 mg/kg/day, i.m.) were administered alone and as co-therapy with the extract of C. colocynthis (25 mg/kg/day, p.o.). Physiological, biochemical and histological examinations were performed to compare the experimental and toxic groups (n = 6) with control group animals. Co-therapy of C. colocynthis with gentamicin protected changes in the body weight, blood urea nitrogen, creatinine clearance, proteins and lactate dehydrogenase excretions. However, a significant rise in serum creatinine and serum uric acid with fall in serum calcium and serum potassium was observed, which were significantly different from control group animals. Necrotic and ruptured tubules were also found abundantly. This study revealed that co-theapy of C. colocynthis with gentamicin for twenty one days, failed to protect renal injury associated by gentamicin in spite of its strong antioxidant properties.

The Biochemistry Section at the Ayub Agricultural Research Institute (AARI) in Faisalabad served as the research hub for our study throughout 2020-21. We delved into the antioxidants and biochemical parameters found in an array of fruits including peach (Prunus persica), grapes (Vitis vinifera), grapefruit (Citrus paradisi), lychee (Litchi chinensis), fig (Ficus carica), plum (Prunus domestica), sweet lime (Citrus limetta), Kuranda (Carissa carandas), kiwi (Actinidia deliciosa), and pear (Pyrus communis). These fruits offer vital nutrition like Vitamin C, ranging from 2.49 in fig to 48.7 in sweet lime, mineral matter is minimum (0.34%) in pear and maximum 6.19% in Lychee, crude fiber was highest (7.26%) in kuranda and lowest (0.82%) in Grapefruit, crude fat was less than 1 in all the fruits except kuranda having crude fat 1.66%. crude protein was also less than one in most of the fruits under study except Fig (2.05%), Plum (3.33%) and Kuranda (1.16%). Total antioxidants were higher in Peach (94.1%) followed by Plum (88.5%), Sweet lime (84.6%) and Fig (83.9%) while lowest value was observed in Kuranda (10.4%). Their adequate consumption not only fortifies the immune system but also serves as a shield against afflictions. Phenols packed abundantly within fruits, stand out as some of the most nutrient-rich components. Total phenols were ranging from 13.8 µg GAE/mL (Grapefruit) to 1182.5 µg GAE/mL (Kuranda). Total sugar lies between 6% (Plum) to 14.3% (Luchee). This paper delves deep into an exhaustive analysis of bioactive compounds, exploring their antioxidant capacities within the most widely cultivated fruits. Our exploration identifies that all the fruits are useful due to one or more nutritious elements. Our findings conclusively advocate the regular consumption of fruits, emphasizing their pivotal role in fostering nutrition and safeguarding against diseases. [ABSTRACT FROM AUTHOR]

The experiment was conducted at Biochemistry Section, Ayub Agricultural Research Institute, Faisalabad Pakistan, during the year 2017-18 to assess the total phenols and antioxidants potential of mango, grapes, banana, guava, mulberry, strawberry, loquat, melon, apricot and phalsa. Fruits samples were obtained from the market of Faisalabad Pakistan, washed, dry at room temperature, extracted their juice/pulp. Thejuice was stored at 4 °C for further analysis. Juice samples were processed for Vitamin-C, pH, TSS, total phenolics and total antioxidant capacity. The results showed that among all the ten fruits vitamin C was higher in mango (24.33 mg/100 mL) followed by phalsa (19.74 mg/100 mL) and Grapes (11.25 mg/100 mL), whereas lower value was observed in melon (3.3 mg/100 mL) and loquat (3.4 mg/100 mL). Vitamin C for other fruits lies between these. Highly acidic pH of fresh juices of all the fruits was observed except in melon (5.82) and guava (5.38) which is less acidic. Percentage of TSS was higher in phalsa, mulberry and mangoes (13.8, 13.5 and 13.9% respectively) and lesser in melon (5.6%). Antioxidant capacity (% DPPH reduction) was found higher in grapes (91.9%) and mango (91.2%) which was statistically at par with melon (90.9%), while lesser value was observed in mulberry (51.6%). Total phenolics were significantly higher in grapes (1266.4 pg GAE/mL), whereas minimum phenolic contents were found in melon (4.1 pg GAE/mL). Therefore, it was concluded that grapes are more nutritious and healthy fruit than other. [ABSTRACT FROM AUTHOR]

In the present study, three different leaf extracts of wild variety of Olea europaea were tested for their phytochemical composition, antioxidant and antimicrobial activities. The antimicrobial activities were determined using the well diffusion method. Phytochemical screening revealed that aqueous, ethanol and ethyl acetate extracts contained alkaloids, flavonoids, steroids, saponins, phenols and tannins while only ethyl acetate fraction was negative for glycosides. Ethanol extract of the plant was observed to have highest contents of alkaloids, flavonoids and phenols. The free radical reducing power assay, DPPH and hydrogen peroxide radical scavenging activities of ethanol extract of O. europaea showed a linear correlation with phenol contents. Ethanol extract was more effective against most of the pathogenic bacteria while aqueous and ethyl acetate extracts were found more potent against most of the fungal strains. The current study revealed that wild variety of O. europaea is as an efficient source of antioxidant and antimicrobial phytochemicals. [ABSTRACT FROM AUTHOR]

Background: Viola canescens Wall. ex. Roxb. exhibits analgesic, antimalarial and antispasmodic activities. It is used folklorically for the treatment of liver diseases, hypertension, malaria and cancer. The current study investigates phytochemical constituents, antioxidant and hepatoprotective activity of solvent extracts of whole plant of Viola canescens. Methods: Phytochemicals, acute toxicity study and antioxidant activity of Viola canescens methanolic extract (VCME), ethyl acetate fraction (EAF), and partially purified EAF (90% EAF and combination of 80% EAF + 20% methanol fraction (EAF + Me) was carried out. Hepatoprotective activity of VCME, EAF (200 and 400 mg/kg body weight) and partially purified EAF (50 mg/kg body weight) was investigated in carbon tetrachloride (CCl4) intoxicated BALB/c mice for 7 days. Membrane stabilization effect was determined by hypotonic solution induced hemolysis while DNA ladder assay was carried out by polyacrylamide gel electrophoresis. Results: Phytochemical screening of VCME showed the presence of alkaloids, phenols, flavonoids, saponins, carbohydrates, tannins and triterpenes. VCME, EAF (at 200 and 400 mg/kg body weight) and partially purified EAF (90% EAF and EAF + Me) at 50 mg/kg body weight significantly reduced the level of ALT, ALP, total bilirubin and restored the level of serum protein in comparison to CCl4 treated group. A significant reduction in malondialdehyde (MDA) and elevation in catalase (CAT) and superoxide dismutase (SOD) level was observed in extract treated animals as compared to CCl4 (p < 0.05). The IC50 values in membrane stabilization potential for VCME, EAF and sodium salicylate were 3.7 ± 0. 11, 3.4 ± 0.15 and 3.2 ± 0.09 mg/ml, respectively. Similarly, CCl4 induced degradation of DNA was counteracted by VCME and EAF. The liver biopsy of mice treated with the solvent extracts showed remarkable restoration of normal histological archeitecture. Conclusions: Viola canescens showed significant hepatoprotective potential due to its antioxidant and membrane stabilization effect. [ABSTRACT FROM AUTHOR]

Background: Cholinesterase inhibition is a vital target for the development of novel and mechanism based inhibitors, owing to their role in the breakdown of acetylcholine (ACh) neurotransmitter to treat various neurological disorders including Alzheimer's disease (AD). Similarly, free radicals are implicated in the progression of various diseases like neurodegenerative disorders. Due to lipid solubility and potential to easily cross blood brain barrier, this study was designed to investigate the anticholinesterase and antioxidant potentials of the standardized essential oils from the leaves and flowers of Polygonum hydropiper. Methods: Essential oils from the leaves (Ph.LO) and flowers (Ph.FO) of P. hdropiper were isolated using Clevenger apparatus. Oil samples were analyzed by GC-MS to identify major components and to attribute the antioxidant and anticholinesterase activity to specific components. Acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) inhibitory potentials of the samples were determined following Ellman's assay. Antioxidant assays were performed using 1,1-diphenyl,2-picrylhydrazyl (DPPH), 2,2-azinobis[3-ethylbenzthiazoline]-6-sulfonic acid (ABTS) and hydrogen peroxide (H2O2) free radical scavenging assays. Results: In the GC-MS analysis 141 and 122 compounds were indentified in Ph.LO and Ph.FO respectively. Caryophylene oxide (41.42 %) was the major component in Ph.FO while decahydronaphthalene (38.29 %) was prominent in Ph.LO. In AChE inhibition, Ph.LO and Ph.FO exhibited 87.00** and 79.66***% inhibitions at 1000 µg/ml with IC50 of 120 and 220 µg/ml respectively. The IC50 value for galanthamine was 15 µg/ml. In BChE inhibitory assay, Ph.LO and Ph.FO caused 82.66*** (IC50 130 µg/ml) and 77.50***% (IC50 225 µg/ml) inhibitions respectively at 1000 µg/ml concentration. In DPPH free radical scavenging assay, Ph.LO and Ph.FO exhibited IC50 of 20 and 200 µg/ml respectively. The calculated IC50s were 180 & 60 µg/ml for Ph.LO, and 45 & 50 µg/ml for Ph.FO in scavenging of ABTS and H2O2 free radicals respectively. Conclusions: In the current study, essential oils from leaves and flowers of P. hydropiper exhibited dose dependent anticholinesterase and antioxidant activities. Leaves essential oil were more effective and can be subjected to further in-vitro and in-vivo anti-Alzheimer's studies. [ABSTRACT FROM AUTHOR]

Fruits of Silybum marianum (L.) Gaernt are the main source of taxifolin derived flavonolignans. Together, these molecules constitute a mixture called silymarin with many useful applications for cosmetic and pharmaceutic industries. Here, a validated method for the separation of the silymarin constituents has been developed to ensure precision and accuracy in their quantification. Each compound was separated with a high reproducibility. Precision and repeatability of the quantification method were validated according to the AOAC recommendations. The method was then applied to study the natural variability of wild accessions of S. marianum. Analysis of the variation in the fruits composition of these 12 accessions from Pakistan evidenced a huge natural diversity. Correlation analysis suggested a synergistic action of the different flavonolignans to reach the maximal antioxidant activity, as determined by cupric ion reducing antioxidant capacity (CUPRAC) and ferric reducing antioxidant power (FRAP) assays. Principal component analysis (PCA) separated the 12 accessions into three distinct groups that were differing from their silymarin contents, whereas hierarchical clustering analysis (HCA) evidenced strong variations in their silymarin composition, leading to the identification of new silybin-rich chemotypes. These results proved that the present method allows for an efficient separation and quantification of the main flavonolignans with potent antioxidant activities. [ABSTRACT FROM AUTHOR]

Ziziphus oxyphylla Edgew (ZO) has been traditionally used for the treatment of hepatic diseases in Pakistan. The current study was carried out to investigate the effectiveness and mechanism underlying hepatoprotective effect of ZO. Hepatoprotective potential of root extracts (crude methanolic extract and subsequent fractions) was investigated against carbon tetrachloride (CCl4) induced hepatotoxicity in BALB/c mice via determination of serum biomarkers, antioxidant enzymes (catalase, superoxide dismutase), lipid peroxidation and histopathology. Membrane stabilizing activity was determined via inhibition of hypotonic solution and heat-induced hemolysis. Crude methanolic extract and chloroform fraction ameliorated CCl4 induced elevated levels of alanine transaminase (ALT), alkaline phosphatase (ALP), total proteins (TP), total bilirubin (TB) and reversed the antioxidants imbalance in terms of catalase (CAT), superoxide dismutase (SOD) and reduced malondialdehyde (MDA) level. Crude methanol extract inhibited hemolysis induced by hypotonic solution and heat. Histopathological examination of liver specimens of extracts administered mice exhibited recovery from CCl4 induced pathological changes. Moreover, CCl4 induced alteration in body and liver weight was countered by the administration of ZO. Compounds betulinic acid (AZO1), ceanothic acid (AZO2) and zizybrenalic acid (AZO3) were isolated from chloroform fraction of the plant. The structures of compounds were deduced using 1D-and 2D-NMR spectroscopy techniques. The results show that Ziziphus oxyphylla roots extract possess antihepatotoxic activity mediated by the protection of anti-oxidant defense system and membrane-stabilizing activity, probably due to the presence of pentacyclic triterpenes. [ABSTRACT FROM AUTHOR]

The pomegranate (Punica granatum L.) is popular for its high medicinal value. It is a common table fruit and whole fruit contains important vitamins, minerals and antioxidants. Four pomegranate varieties (Pearl, Golden, Sultan, and Qandhari) were analyzed in the Biochemistry laboratory of Post Harvest Research Centre, Ayub Agricultural Research Institute (AARI), Faisalabad Pakistan during the year 2021-2022. The results showed that juice carried a remarkably higher value of TSS (total soluble solids) ranging from 12.4% (Pearl) to 17.2 % (Sultan), total invert sugars were minimum (11.8%) in Golden and maximum (14.3%) in Sultan. The value of vitamin-C varies from 10.6 mg/100 mL (Qandhari) to 11.5 mg/100 mL (Pearl) and acidity ranges from 0.8% (Sultan) to 1.9% (Pearl). Sultan showed a higher value for total phenols and antioxidants (1111µg GAE L-1 and 43.5% DPPH inhibition respectively) and lowest in Pearl (1022µg GAE L-1 and 27.7% respectively). Peel powder of different varieties carry a different percentage of crude fat and crude fiber but Sultan is statistically at par with Qandhari but significantly higher than pearl. Ash/mineral matter was significantly different in different varieties. Sultan was better than other varieties having ash contents of 3.67%. The lowest value was observed in Golden (2.7%). Keeping in view the current results Sultan was better than Pearl and Golden and comparable to Qandhari. [ABSTRACT FROM AUTHOR]

Purpose: To synthesize copper oxide (CuO) nanoparticles using a ecofriendly technique and evaluate their antimicrobial, antioxidant and photo-catalytic dye degradation potentials. Methods: A superficial method (solution combustion method) was employed for the synthesis of copper oxide nanoparticles from an aqueous extract of Abutilon indicum. The CuO nanoparticles were characterized using x-ray diffraction (XRD), energy-dispersive x-ray spectroscopy (EDX), scanning electron microscope (SEM) and ultraviolet-visible (UV-Vis) spectroscopic techniques. The antimicrobial activity of the CuO nanoparticles was determined by agar well diffusion method, while their antioxidant properties were assessed by DPPH radical scavenging, ferric reducing antioxidant power (FRAP), total antioxidant, ferric thiocyanate (FTC) and total phenolic content (TPC) assays. The photo-catalytic degradation activity of synthesized CuO nanoparticles was assessed by the degradation of Acid Black 210 (AB) dye under sunlight irradiation. Results: XRD, EDX and SEM results confirmed successful synthesis of CuO nanoparticles, with hexagonal, wurtzite and sponge crystal structure. Photo-catalytic data revealed that the nanoparticles are a good catalyst for effective degradation of Acid Black 210. The nanoparticles also exhibited remarkable antioxidant activity, with IC50 and FRAP values ranging from 40 ± 0.23 to 84 ± 0.32 μg/ml, and 0.65 ± 0.01 to 9.10 ± 0.21 Trolox equivalent/mL, respectively. Significant bactericidal activity was manifested by the CuO nanoparticles against Klebsiella and Bacillus subtilis with zone of inhibition of 14 ± 0.05 and 15 ± 0.11 mm, respectively. Conclusion: The synthesized CuO nanoparticles exhibit antibacterial and antioxidant potential, indicating that they are good candidates for future therapeutic applications. [ABSTRACT FROM AUTHOR]