Antioxidant and Hypoglycemic Potential of Phytogenic Selenium Nanoparticle- and Light Regime-Mediated In Vitro Caralluma tuberculata Callus Culture Extract.

In vitro plant cultures have emerged as a viable source, holding auspicious reservoirs for medicinal applications. This study aims to delineate the antioxidant and hypoglycemic potential of phytosynthesized selenium nanoparticle (SeNP)- and light stress-mediated in vitro callus cultures of Caralluma tuberculata extract. The morphophysicochemical characteristics of biogenic SeNPs were assessed through a combination of analytical techniques, including UV-visible spectrophotometry, scanning electron microscopy, energy-dispersive X-rays, Fourier transform infrared spectrometry, and zeta potential spectroscopy. The antioxidative potential of the callus extract 200 and 800 µg/mL concentrations was assessed through various tests and exhibited pronounced scavenging potential in reducing power (26.29%), ABTS + scavenging (42.51%), hydrogen peroxide inhibition (37.26%), hydroxyl radical scavenging (40.23%), and phosphomolybdate (71.66%), respectively. To inspect the hypoglycemic capacity of the callus extract, various assays consistently demonstrated a dosage-dependent relationship, with higher concentrations of the callus extract exerting a potent inhibitory impact on the catalytic sites of the alpha-amylase (78.24%), alpha-glucosidase (71.55%), antisucrase (59.24%), and antilipase (74.26%) enzyme activities, glucose uptake by yeast cells at 5, 10, and 25 mmol/L glucose solution (72.18, 60.58 and 69.33%), and glucose adsorption capacity at 5, 10, and 25 mmol/L glucose solution (74.37, 83.55, and 86.49%), respectively. The findings of this study propose selenium NPs and light-stress-mediated in vitro callus cultures of C. tuberculata potentially operating as competitive inhibitors. The outcomes of the study were exceptional and hold promising implications for future medicinal applications.

Exposure of Caralluma tuberculata to biogenic selenium nanoparticles as in vitro rooting agent: Stimulates morpho-physiological and antioxidant defense system.

The commercial-scale production of Caralluma tuberculata faces significant challenges due to lower seed viability and sluggish rate of root growth in natural conditions. To overcome these obstacles, using phyto-mediated selenium nanomaterials as an in vitro rooting agent in plant in vitro cultures is a promising approach to facilitate rapid propagation and enhance the production of valuable therapeutic compounds. This study aimed to investigate the impact of phytosynthesized selenium nanoparticles (SeNPs) on the morphological growth attributes, physiological status, and secondary metabolite fabrication in in vitro propagated Caralluma tuberculata. The results demonstrated that a lower dose of SeNPs (100 µg/L) along with plant growth regulators (IBA 1 mg/L) had an affirmative effect on growth parameters and promoted earliest root initiation (4.6±0.98 days), highest rooting frequency (68.21±5.12%), number of roots (6.3±1.8), maximum fresh weight (710±6.01 mg) and dry weight (549.89±6.77 mg). However, higher levels of SeNPs (200 and 400 µg/L) in the growth media proved detrimental to growth and development. Further, stress caused by SeNPs at 100 µg/L along with PGRs (IBA 1 mg/L) produced a higher level of total chlorophyll contents (32.66± 4.36 µg/ml), while cultures exposed to 200 µg/L SeNPs alone exhibited the maximum amount of proline contents (10.5± 1.32 µg/ml). Interestingly, exposure to 400 µg/L SeNPs induced a stress response in the cultures, leading to increased levels of total phenolic content (3.4 ± 0.052), total flavonoid content (1.8 ± 0.034), and antioxidant activity 82 ± 4.8%). Furthermore, the combination of 100 µg/L SeNPs and plant growth regulators (1 mg/L IBA) led to accelerated enzymatic antioxidant activities, including superoxide dismutase (SOD = 4.4 ± 0.067 U/mg), peroxidase dismutase (POD = 3.3 ± 0.043 U/mg), catalase (CAT = 2.8 ± 0.048 U/mg), and ascorbate peroxidase (APx = 1.6 ± 0.082 U/mg). This is the first report that highlights the efficacy of SeNPs in culture media and presents a promising approach for the commercial propagation of C. tuberculata with a strong antioxidant defense system in vitro.

Purification and characterization of novel isoforms of the polyphenol oxidase from Malus domestica fruit pulp.

Polyphenol oxidases (PPOs), belong to the group of oxidoreductases that are copper containing enzymes and are responsible for plant browning. PPOs are extensively distributed in plant kingdom and can oxidize wide range of aromatic compounds of industrial importance. The aim of this study was purification and characterization of PPO isoforms from the fruit pulp of Golden delicious apple. High performance liquid chromatography was used to purify the two novel isoforms of PPO and further their molecular weights (45 and 28 kDa) were determined using sodium dodecyl sulfate polyacrylamide gel electrophoresis. The purified isoforms have optimum pH (6.5), optimum temperature (40°C), the Vmax (4.45 µM/min) and Km (74.21 mM) with catechol substrate. The N-terminal microsequences of both PPO isoforms were determined using a pulse liquid protein sequencer and found to be AKITFHG (28 kDa) and APGGG (45 kDa). Polyphenol oxidases are efficiently used in the pharmaceutical, paper and pulp, textiles and food industries. Recently, the PPOs have been used for bioremediation and in the development of biosensors.

Calcium based siRNA coating: a novel approach for knockdown of HER2 gene in MCF-7 cells using gold nanoparticles.

Surface functionalization of nanoparticles (NPs) for therapeutic siRNA delivery into cancer cells has gained interest. The present study was designed for surface functionalization of gold nanoparticles (AuNPs) for efficient siRNA delivery and knockdown in cancer cells. In order to achieve this objective, AuNPs were coated with HER2-siRNA in the presence of 11-mercaptoundecanoic acid (11-MUA), calcium chloride (CaCl2) and polyethyleneimine (PEI) in alternate charge bearing successive layers. MCF-7 cells were cultured and transfected with fabricated assembly of AuNPs. Cytotoxicity analysis revealed that the half inhibitory concentration (IC50) for the formulation was 45.35 nM  . Total RNA was isolated from transfected cells, reverse transcribed into complementary DNA (cDNA) and real-time polymerase chain reaction (RT-PCR) was performed. The RT-PCR based delta-delta Ct analysis in treated cells revealed a significant 18.94 times decrease (p<0.001) in the expression of HER2 gene standardized with ACTB housekeeping gene as compared to untreated cells, which makes this formulation a potent approach for siRNA delivery and  gene knockout.

Formulation and evaluation of antioxidant and antityrosinase activity of Polygonum amplexicaule herbal gel.

Medicinal plants are long been used for pharmaceutical and cosmetic industry. Among medicinal plants, Polygonum amplexicaule of family polygonaceae has traditional use in medicines and skin care. P. amplexicaule belongs to genus Polygonum that contains several important phytochemicals and considered as a rich source of antioxidants. The present study was designed to formulate herbal gel containing P. amplexicaule extract and evaluate its different physical properties as well as antioxidants and antityrosinase activities. Chitosan gel base was used as gelling agent and different gel formulations were prepared by different concentrations of extracts and polymers. Physical properties like pH, colour, odour, appearance and homogeneity, spreadability, extrudability and stability were optimized and analysed. A stable gel formulation containing 1% chitosan gel base and 5% plant extract was prepared that showed good appearance and homogeneity, easily spread ability and excellent extrudability. This gel formulation was tested for antioxidant and skin whitening properties by DPPH free radical scavenging assay and tyrosinase inhibition assay respectively and ascorbic acid was used as reference standard. DPPH scavenging activity with an IC50 value of 0.446 mg/mL and tyrosinase inhibition activity with an IC50 value of 0.805 mg/mL was observed and results indicated that this herbal gel formulation has a good potential for cosmetic use.

In vitro anticancer activities of Withania coagulans against HeLa, MCF-7, RD, RG2, and INS-1 cancer cells and phytochemical analysis.

BACKGROUND: The Pakistani Salt Range has a rich floral diversity including Withania coagulans from the Solanaceae family. METHODS: The crude methanolic extracts of the root, leaf, leaf stalk, and fruit of this plant were screened for their cytotoxic activity against human (HeLa, MCF-7, RD) and rat (RG2 and INS-1) cancer cell lines at 20 µg/mL and compared to methotrexate. The IC50 values indicated that leaf stalk and fruit extracts exert an 80% or higher cytotoxic activity against all cell lines at 24 hours. RESULTS: The leaf stalk extract showed the highest cytotoxic efficacy against all tested cell lines, with IC50 values ranging from 0.96 ± 0.01 µg/mL to 4.73 ± 0.05 µg/mL followed by the fruit extract with IC50 values of 0.69 ± 0.01-6.69 ± 0.06 µg/mL after 48-72 hours incubation. The leaf stalk and seed extracts were analyzed for polyphenols and flavonoids using RP-HPLC. The total flavonoid content (TFC) was calculated for all tested samples, and the highest TFC was recorded for the root extract (394.34 ± 1.26 µg/g). The total phenolic content (TPC) was found in the seed extract (307.86 ± 9.42 µg/g) of W. coagulans. The highest contents of myricetin (358.46 ± 2.91 µg/g) were noted in the leaf extract, and highest quercetin was recorded in the seed extract (21.43 ± 0.13 µg/g). The highest gallic acid concentration (83.62 ± 0.71 µg/g) was recorded in leaf stalk extract and p-hydroxybenzoic acid in the seed extract (157.46 ± 1.43 µg/g). CONCLUSION: The present study gives a scientific insight and comparative analysis of various plant parts in this medicinally important plant species from the Salt Range of Pakistan against both human and rat cancer cells.

Evaluation of antioxidant potential and HPLC based identification of phenolics in Polygonum amplexicaule extract and its fractions.

There is a growing interest for the plant-based medicines in pharmaceutical industry. Plant derived Antioxidants have gained huge importance regarding their medicinal value. The present study was designed to establish pharmaceutical value of Polygonum amplexicaule for their antioxidant activity using shoot, leaf and rhizome crude methanolic extract along with their n-butanolic, ethanolic, ethyl acetate and aqueous fractions. DPPH assay was used to assess antioxidants, which shows the maximum activity by crude methanolic extract of leaves (CMEL) having IC(50) 1.03 µg/ml where all other fractions showed IC(50) in a range of 1.03-58.2 µg/mL. The DNA plasmid protection assay showed that 10 ppm and 100 ppm concentrations of crude methanolic extracts (rhizome and leaf), aqueous fractions (shoot and leaf extract), n-butanolic fractions (shoot and leaf extract) and ethanolic fraction (rhizome extract) have DNA protection properties. TLC and HPLC based Identification of different antioxidants present in shoot, leaf and rhizome crude extracts and their fractions showed the presence of gallic acid, quercetin, catechin, caffeic acid, rutin, myricetin and kaempferol. This study suggested that this plant have high content of antioxidants, which needs to be investigated further for their medicinal and/cosmaceutical applications.