Hepatoprotective potential of coconut inflorescence sap against paracetamol induced toxicity in hep G2 cell lines.

Coconut Inflorescence Sap (CIS) is the sweet, oyster-white colored, non-fermented juice obtained from the immature inflorescence of the Coconut tree. Acetaminophen (N-acetyl-p-aminophenol, or paracetamol) is one of the most frequently used drugs worldwide as an antipyretic or analgesic. HepG2 cell lines were used as an experimental model for studying in vitro hepatotoxicity induced by Paracetamol. The present study aims to identify biologically active compounds of CIS using LCMS analysis and to elucidate the ameliorative potential of CIS in alleviating paracetamol-induced hepatotoxicity. LC-MS analysis revealed the presence of 17 bioactive compounds. HepG2 cells were pretreated with Paracetamol (20 mM) for inducing toxicity, and Silymarin at a concentration of 50 µg/ml was used as a standard drug. The morphological analysis and MTT assay showed effective recovery from toxicity in cells treated with CIS in a dose-dependent manner. CIS at 25 µg/ml potentially showed the highest percentage of inhibitory activity against the toxicity induced by paracetamol. The treatment with paracetamol significantly increased the indicators of liver toxicity - LDH, SGOT, SGPT, and Glut.S Transferase in the media.CIS administration also increased the total protein levels, SOD, and Catalase activity. The morphological analysis, MTT assay, cytocompatibility studies, determination of enzymatic activities, etc., confirms the significant hepatoprotective efficacy of CIS.

Metagenomic Investigation Uncovers Presence of Probiotic-Type Microbiome in Kalparasa® (Fresh Unfermented Coconut Inflorescence Sap).

The phloem sap tapped from unopened inflorescence (spadix) of coconut palm using a novel collecting device, "coco-sap chiller," has been branded Kalparasa® (henceforth as Kalparasa in the text) to distinguish its properties not found in sap harvested by traditional methods. To know its hitherto unidentified microbiome profile, we employed high-throughput sequencing to uncover the bacteriome and mycobiome in fresh and 12-h fermented samples. Fresh Kalparasa had a pH of 7.2, which dropped to 4.5 after 12 h, signifying fermentation of the sap. Diversity analysis indicated fresh Kalparasa having higher bacterial species than the fermented one. Contrary to this, fresh sap had lower fungal/yeast diversity than the fermented sample. Fresh Kalparasa had relatively higher abundance of probiotic-type Leuconostoc genus followed by equal proportions of Gluconobacter, Acetobacter, and Fructobacillus. The 12-h fermented Kalparasa showed a significant increase in Gluconobacter with a sharp decrease in Leuconostoc. Mycobiome data revealed fresh Kalparasa to be preponderant in Saccharomyces and Hanseniaspora genera of yeasts while the fermented sap had higher representation of Hanseniaspora and Cortinarius and lesser Saccharomyces. This suggested that the fermentation of Kalparasa was probably driven by symbiotic culture of bacteria and yeasts (SCOBY), particularly acetic acid bacteria and non-Saccharomyces yeasts. The bacteriome-function predictions highlighted the enrichment of glycerophospholipid, ABC transporters, purine, and pyrimidine metabolisms. Based on our findings, Kalparasa containing large population of Leuconostoc mesenteroides, Fructobacillus fructosus, Saccharomyces cerevisiae, and Hanseniaspora guilliermondii can be promoted as a healthy "unfermented" plant edible food containing live probiotic-type microbiome during its consumption.

Reaction kinetics of physico-chemical attributes in coconut inflorescence sap during fermentation.

The study on fermentation kinetics of the coconut inflorescence sap is important to understand its shelf life at different storage conditions and to develop suitable value added products. The coconut inflorescence sap collected by using in-house developed coco-sap chiller device is called Kalparasa. The fermentation characteristics of Kalparasa were investigated at every 1-h interval under ambient (31 ± 2 °C) and refrigerated (5 ± 1 °C) storage conditions. The results reveal that pH of the sap and total sugar content decline rapidly under ambient conditions than under refrigerated conditions. Acidity, turbidity, and reducing sugar content significantly (p < 0.001) increases for the sap stored under ambient conditions. The reaction rate constant (k) of the vitamin C and total sugar degradation increases with the atmospheric fermentation. The degradation kinetics of vitamin C and total sugar in Kalparasa during natural fermentation (ambient condition) follow second-order equation whereas the reducing sugar follows the first-order equation.

Neera, a non-fermented traditional drink from coconut spadix restores the redox status in sodium fluoride intoxicated mice.

OBJECTIVES: Neera, non-fermented coconut inflorescence sap (NFCIS) and its fermented form, toddy (FCIS) are the two well-known traditional drinks in South India. Both the saps show high rejuvenation effect and NFCIS is used for the curing of bronchial suffocation, anemia, tuberculosis and piles in traditional medicine. But, a few scientific studies have been reported on their health benefits so far. Presently, the antioxidant effect of both non-fermented (NFCIS) and fermented form (FCIS) of coconut inflorescence were analyzed in experimental animals. METHODS: The free radical scavenging property of FCIS and NFCIS was analyzed in vitro. The effect of these saps on mitigating sodium fluoride (NaF) deteriorated redox status was evaluated in mice. RESULTS: NFCIS exhibited high antioxidant activity than its fermented form. NFCIS reduces metal ions and scavenge hydroxyl and DPPH radicals with IC50 values 6.5 ± 1.9 and 44 ± 3.14 µL/mL, respectively. Supplementation of NFCIS for 14 days increased SOD, CAT and GPx activities and GSH level in liver by 51.67, 52, 27.88 and 35.77%, respectively against NaF induced decline with a concomitant decrease in lipid peroxidation to 40.76%. Saps rich in minerals indicate pharmaceutical and nutritional value. CONCLUSION: The study revealed the antioxidant efficacy of non-alcoholic natural drink, Neera and recommends an alternative for synthetic carbonated soft drinks. The regular consumption of Neera may protect the body from various chronic diseases especially where the oxidative stress played as a key role.

Ameliorative effect of Neera, nonfermented coconut inflorescence sap, on cisplatin-induced renal toxicity by abating oxidative stress.

Objectives Neera, nonfermented coconut inflorescence sap (NFCIS) from unopened spadix of Cocos nucifera L., is a well-known traditional beverage. But, scientific reports on its health benefits are limited. NFCIS is reported to exhibits free radical scavenging activity, and its chemical composition is found promising. In the present study, the effect of NFCIS on alleviating cisplatin-induced nephrotoxicity was analyzed in mice. Methods The renal toxicity was induced by cisplatin (16 mg/kg b.wt. ip) in Swiss albino mice. The antioxidant activity of NFCIS was evaluated by nitric oxide radical scavenging assay and phorbol-12-myristate-13-acetate-induced superoxide radical generation in mice peritoneal macrophages. Total polyphenolic content of sap was determined using Folin-Ciocalteu reagent. The phytochemicals present in NFCIS was identified using Fourier transform infrared (FT-IR) spectroscopy. Results NFCIS was found to scavenge nitric oxide (NO) radicals (IC50 = 32 ± 2.47 µL/mL) and shown to inhibit superoxide (SO) generation (53.5 ± 2.1%) in macrophages. High polyphenolic content (193 µg gallic acid/mL) was determined in the sap. The FT-IR spectrum of NFCIS revealed the presence of several phytochemicals indicate its pharmaceutical and nutritional value. Cisplatin-induced hike in urea, creatinine and lipid peroxidation was significantly decreased to 65.16, 87.74 and 53.41% by NFCIS, respectively. Hb (42.37%) and total count (72.81%) were also found to be increased. Additionally, the activity of antioxidant enzymes superoxide dismutase, catalase, glutathione peroxidase and reduced glutathione was enhanced to 53.06, 40, 52.22 and 38.49%, respectively. Conclusions Results indicate that NFCIS effectively alleviates cisplatin-mediated renal toxicity by its antioxidant activity.

Facile coconut inflorescence sap mediated synthesis of silver nanoparticles and its diverse antimicrobial and cytotoxic properties.

Green synthesis of nanoparticles (NPs) involves the use of diverse extracts of biological origin as substrates to synthesize NPs and can overcome the hazards associated with chemical methods. Coconut inflorescence sap, which is unfermented phloem sap obtained by tapping of coconut inflorescence, is a rich source of sugars and secondary metabolites. In this study, coconut inflorescence sap was used to synthesize silver NPs (AgNPs). We have initially undertaken metabolomic profiling of coconut inflorescence sap from West Coast Tall cultivar to delineate its individual components. It was found to comprise of 64% secondary metabolites, 9% sugars, 12% lipids/fats and 9% peptides in positive mode, whereas in the negative mode, it was 33, 20, 9 and 11%, respectively. The concentration of silver nitrate, inflorescence sap and incubation temperature for the synthesis of AgNPs were optimized. Incubating the reaction mixture at 40 °C was found to enhance AgNP synthesis. The AgNPs synthesized were characterized using UV-visible (UV-Vis) spectrophotometry, X-Ray Diffraction (XRD), Fourier Transform Infrared spectroscopy (FTIR) and Transmission Electron Microscopy (TEM). The particles were crystalline in nature and the bulk of the particles were spherical with smooth (thin) shell and poly-dispersed with a diameter ranging from 10 nm to 30 nm. Antimicrobial property of AgNPs was tested in tissue culture of arecanut (Areca catechu L.) where bacterial contamination (Bacillus pumilus) was a frequent occurrence. A significant reduction in the contamination was observed when plantlets were treated with aqueous solutions of AgNPs. Notably, treatment with AgNPs did not affect the growth and development of the arecanut plantlets. Antimicrobial properties of AgNPs synthesized from inflorescence sap were also evaluated in human pathogenic bacteria viz., Escherichia coli ATCC 25922; Salmonella Typhimurium ATCC 14028 and Vibrio parahaemolyticus AQ4037. The antibacterial action was confirmed by determining the production of reactive oxygen species (ROS) and protein leakage studies. Cytotoxicity of AgNPs was quantified in HeLa cells. The viability (%) of HeLa cells declined significantly at 10 mg L-1 concentration of AgNP and complete mortality was observed at a concentration of 60 mg L-1. The study concludes that unfermented inflorescence sap, with above neutral pH, serves as an excellent reducing agent to synthesize AgNPs from Ag+.

Nephro-protective effect of a novel formulation of unopened coconut inflorescence sap powder on gentamicin induced renal damage by modulating oxidative stress and inflammatory markers.

Fresh oyster white translucent sap obtained from the tender unopened inflorescence of coconut trees (Cocos nucifera) is identified to have great health benefits. Drug induced Nephrotoxicity is one of the major causes of renal damage in present generation. As a therapeutic agent, gentamicin imparts direct toxicity to kidney, resulting in acute tubular necrosis, glomerular and tubulointerstitial injury, haemodynamically mediated damage and obstructive nephropathy.There exists an increasing demand for safe and natural agents for the treatment and/or preventionofchronic nephrotoxicity and pathogenesis of kidney diseases. Our study shows the nephro protective/curing effect of a novel powder formulation of micronutrient enriched, unfermented coconut flower sap (CSP). The study was performed on adult male Wistar rats. The animals were grouped into three and treated separately with vehicle, gentamicin and gentamicin+CSP for 16days. Initially, gentamicin treatment significantly (p<0.05)reduced thelevels of antioxidant enzymes (SOD, CAT, GPx) and GSH and increased (p<0.05) the levels of creatinine, uric acid, urea, inflammatory markers (nitrite, IL-6, TNF- α, iNOS) and lipid peroxidation. Supplementation of coconut flower sap powder showed significant (p<0.05) reversal of all these biochemical parameters indicating an effective inhibition of the pathogenesis of nephrotoxicity and kidney disease.