Elucidation of the interaction of apocarotenoids with calf thymus DNA by biophysical techniques and in vitro study in MCF-7 cells to explore their potential in cancer therapy

Objective(s): DNA is one of the targets of cancer-therapeutic small molecules. Cisplatin, a DNA intercalator, is one of the first-line drugs in the cancer chemo regimen which comes with health-compromising side effects during chemotherapy. The synergistic effect of natural molecules with cisplatin can help to potentiate its anti-cancer efficacy and decrease its negative effect on health. Here, we report the interaction of cisplatin with calf thymus-DNA (ct-DNA) in combination with natural molecules like apocarotenoids which are reported for their therapeutic properties.Materials and Methods: The combinatorial effect of apocarotenoids on ct-DNA was explored through various biophysical techniques such as UV-Visible spectroscopy, circular dichroism studies, DNA melt curve analysis, viscosity measurements, and an in vitro study in MCF-7 cells by cell cycle analysis.Results: UV-Visible spectroscopy studies suggest apocarotenoids and their combination shows a non-intercalative mode of binding. Circular dichroism analysis showed no major changes in DNA form during the interaction of DNA with apocarotenoids and their respective combinations with cisplatin, which is suggestive of the groove-binding mode of apocarotenoids. DNA melt curve analysis showed a decrease in the intensity of the fluorescence for apocarotenoids with cisplatin which indicates the possibility of DNA interaction through groove binding. Viscosity studies suggested a groove binding mode of interaction of ct-DNA with apocarotenoids and their combination as there was minimal change in the viscosity measurements. The in vitro analysis exhibits that the apocarotenoids and their combination have a considerable effect on DNA synthesis.Conclusion: This study provides a better perspective on the possible mode of interaction between ct-DNA and natural molecules along with cisplatin

DESIGN, 23 FACTORIAL OPTIMIZATION AND IN VITRO–IN VIVO PHARMACOKINETIC EVALUATION OF ROSUVASTATIN CALCIUM LOADED POLYMERIC NANOPARTICLES

Objective: The objective is to incorporate low bioavailable Rosuvastatin Calcium (20%) into polymeric nanoparticles (PNs) to improve its biopharmaceutical properties of Rosuvastatin Calcium. Methods: The PNs were prepared by solvent evaporation method by applying 23 factorial designs. The formulated PN are investigated for particle size (PS) and shape, zeta potential (ZP), polydispersity index (PI) and entrapment efficiency (EE), in vivo pharmacokinetic. Results: Among 8 formulations, PN7 shows least PS of 159.9±16.1 nm, which enhance the dissolution, surface area and permeability; ZP of-33.5±1.54 mV, which shows good stability; PI of 0.587±0.16 shows monodisperse distribution pattern; high EE of about 94.20±2.46 %; percentage yield of 96.80±2.08 %; maximum in vitro drug release of about 96.54±2.02 % at 24 h with controlled and predetermined release pattern. PN7 drug release obeys zero-order release kinetics, non-fickian diffusion mechanism with r2 value>0.96 and release exponent ‘n’ value falls between 0.5-0.8 for peppas kinetic model i.e., the mechanism of drug diffusion is based on polymer relaxation. In vivo pharmacokinetic studies illustrate enhance in AUCo-α in mg/ml, which proves a significant enhancement of bioavailability of Rosuvastatin Calcium by PNs. Conclusion: This PN shows a significant enhancement of bioavailability by minimizing the dose-dependent adverse side effects of rosuvastatin calcium

Fruit ripening: dynamics and integrated analysis of carotenoids and anthocyanins

Background: Fruits are vital food resources as they are loaded with bioactive compounds varying with different stages of ripening. As the fruit ripens, a dynamic color change is observed from green to yellow to red due to the biosynthesis of pigments like chlorophyll, carotenoids, and anthocyanins. Apart from making the fruit attractive and being a visual indicator of the ripening status, pigments add value to a ripened fruit by making them a source of nutraceuticals and industrial products. As the fruit matures, it undergoes biochemical changes which alter the pigment composition of fruits. Results: The synthesis, degradation and retention pathways of fruit pigments are mediated by hormonal, genetic, and environmental factors. Manipulation of the underlying regulatory mechanisms during fruit ripening suggests ways to enhance the desired pigments in fruits by biotechnological interventions. Here we report, in-depth insight into the dynamics of a pigment change in ripening and the regulatory mechanisms in action. Conclusions: This review emphasizes the role of pigments as an asset to a ripened fruit as they augment the nutritive value, antioxidant levels and the net carbon gain of fruits; pigments are a source for fruit biofortification have tremendous industrial value along with being a tool to predict the harvest. This report will be of great utility to the harvesters, traders, consumers, and natural product divisions to extract the leading nutraceutical and industrial potential of preferred pigments biosynthesized at different fruit ripening stages

The role of photosynthesis related pigments in light harvesting, photoprotection and enhancement of photosynthetic yield in planta.

Photosynthetic pigments are an integral and vital part of all photosynthetic machinery and are present in different types and abundances throughout the photosynthetic apparatus. Chlorophyll, carotenoids and phycobilins are the prime photosynthetic pigments which facilitate efficient light absorption in plants, algae, and cyanobacteria. The chlorophyll family plays a vital role in light harvesting by absorbing light at different wavelengths and allowing photosynthetic organisms to adapt to different environments, either in the long-term or during transient changes in light. Carotenoids play diverse roles in photosynthesis, including light capture and as crucial antioxidants to reduce photodamage and photoinhibition. In the marine habitat, phycobilins capture a wide spectrum of light and have allowed cyanobacteria and red algae to colonise deep waters where other frequencies of light are attenuated by the water column. In this review, we discuss the potential strategies that photosynthetic pigments provide, coupled with development of molecular biological techniques, to improve crop yields through enhanced light harvesting, increased photoprotection and improved photosynthetic efficiency

Ethnoveterinary medicine of the Shervaroy Hills of Eastern Ghats, India as alternative medicine for animals

The Eastern Ghats of India is well known for its wealth of natural vegetation and Shervaroy is a major hill range of the Eastern Ghats of Tamil Nadu. Ethnomedicinal studies in the Eastern Ghats of Tamil Nadu or the Shervaroy Hills have been carried out by various researchers. However, there is not much information available on ethnoveterinary medicine in the Eastern Ghats of India. The aim of this study was to examine the potential use of folk plants as alternative medicine for cattle to cure various diseases in the Shervaroy Hills of the Eastern Ghats. Based on interactions with traditional medicine practitioners, it has been observed that a total of 21 medicinal plants belonging to 16 families are used to cure various diseases such as mastitis, enteritis, arthritis, stomatitis, salivation from the mouth, wounding, and conjunctivitis in animals. It has been observed that the traditional knowledge of ethnoveterinary medicine is now confined only among the surviving older people and a few practitioners in the tribal communities of the Shervaroy Hills. Unfortunately, no serious attempts have been made to document and preserve this immense treasure of traditional knowledge

Biologically active recombinant human erythropoietin expressed in hairy root cultures and regenerated plantlets of Nicotiana tabacum L.

Hairy root culture is a potential alternative to conventional mammalian cell culture to produce recombinant proteins due to its ease in protein recovery, low costs and absence of potentially human pathogenic contaminants. The current study focussed to develop a new platform of a hairy root culture system from Nicotiana tabacum for the production of recombinant human EPO (rhEPO), which is regularly produced in mammalian cells. The human EPO construct was amplified with C-terminal hexahistidine tag from a cDNA of Caco-2 cells. Two versions of rhEPO clones, with or without the N-terminal calreticulin (cal) fusion sequence, were produced by cloning the amplified construct into gateway binary vector pK7WG2D. Following Agrobacterium rhizogenes mediated transformation of tobacco explants; integration and expression of constructs in hairy roots were confirmed by several tests at DNA, RNA and protein levels. The amount of intracellular rhEPO from hairy root cultures with cal signal peptide was measured up to 66.75 ng g-1 of total soluble protein. The presence of the ER signal peptide (cal) was essential for the secretion of rhEPO into the spent medium; no protein was detected from hairy root cultures without ER signal peptide. The addition of polyvinylpyrrolidone enhanced the stabilization of secreted rhEPO leading to a 5.6 fold increase to a maximum concentration of 185.48 pg rhEPOHR g-1 FW hairy root cultures. The rhizo-secreted rhEPO was separated by HPLC and its biological activity was confirmed by testing distinct parameters for proliferation and survival in retinal pigment epithelial cells (ARPE). In addition, the rhEPO was detected to an amount 14.8 ng g-1 of total soluble leaf protein in transgenic T0 generation plantlets regenerated from hairy root cultures with cal signal peptide

Recent advances in extraction methodologies for the valorization of mango peel wastes

ABSTRACTMango is an important tropical edible fruit having economic importance, which is cultivated mainly in India (36.6%). It contains various macronutrients, micronutrients, antioxidants, and other bioactive compounds. It is consumed in fresh form or processed into different products namely pulp, juice, puree, pickle, jam, and nectar. It has been predicted mango processing will increase considerably reaching USD 1.8 billion in 2029.] The by-products generated during the processing of mangoes are peel, pomace, seed, and kernel which constitute 25–40% of fresh fruit. This review article describes the nutritional composition of mango peels and also provides detailed insights into different extraction methodologies for value-added compounds. This review also explores the available literature reports that prove mango peels are an excellent source of carotenoids, pectin, phenolic compounds, and volatile aroma compounds. Mango peels contain significantly higher amounts of minerals compared to pulp. It possesses antioxidant, antimicrobial, cardioprotective, anti-diabetic, and anti-cancer properties. This article emphasizes the advantages of green extraction methodologies like ultrasound or microwave-assisted deep eutectic solvents compared to conventional extraction methods. The sustainable valorization of mango peels generated during processing can be economical as well as environmentally feasible

Genomic DNA and transcript analysis of hairy root cultures.

<p>(A) Southern blot analysis was performed using the genomic DNA of transformed hairy root clones. (B) RT-PCR analysis of <i>EPO</i> mRNA in hairy root cultures ^pK7cal/rh<i>EPO</i>^HR and ^pK7rh<i>EPO</i>^HR. The presence of mRNA for tobacco <i>β actin</i> gene is shown as an internal control for transcript analysis.</p