Rohit Arora

Evaluating Extraction Conditions of Glucosinolate Hydrolytic Products from Seeds of Eruca sativa (Mill.) Thell. Using GC-MS

UNLABELLEDnGlucosinolates and their hydrolytic products form an important class of plant secondary metabolites involved in various plant defense-linked mechanisms. The successful isolation of particular glucosinolate hydrolytic products is limited by a number of factors like understanding the parent glucosinolate moiety, solubility, and stability under different drying conditions. The extraction protocols currently available were modified to achieve both an increased yield as well as an increased number of hydrolytic products. Eruca sativa (Mill.) Thell. (called arugula in the U.S.A.), a rich source of varied glucosinolates, was used for the standardization of different extraction protocols. We exploited the volatile nature of the glucosinolates and developed a method that not only enhanced the yield of glucosinolate hydrolytic products, but also reduced undesired compounds. Among all the tested protocols, hydrodistillation using Clevenger apparatus was judged as the best protocol, which was evident from an enhanced yield as well as an increased number of hydrolytic products when compared to the other methods as monitored by gas chromatography-mass spectrometry.nnnPRACTICAL APPLICATIONnGlucosinolate hydrolytic products are important volatile metabolites that are difficult to extract. The different conditions, such as extraction method, solvent, and dryingmethods, are responsible for successful extractions. An improved extraction method will help in a better isolation of these valuable compounds, which may then be used for different biological activities such as anticancer, antimutagenic, bioherbicidal, antimicrobial, antigenotoxic, and antitumor activities.

Hepatic Dysfunction Induced by 7, 12-Dimethylbenz(α)anthracene and Its Obviation with Erucin Using Enzymatic and Histological Changes as Indicators

The toxicity induced by 7, 12-dimethylbenz(α)anthracene (DMBA) has been widely delineated by a number of researchers. This potent chemical damages many internal organs including liver, by inducing the production of reactive oxygen species, DNA-adduct formation and affecting the activities of phase I, II, antioxidant and serum enzymes. Glucosinolate hydrolytic products like isothiocyanates (ITCs) are well known for inhibiting the DNA-adduct formation and modulating phase I, II enzymes. Sulforaphane is ITC, currently under phase trials, is readily metabolized and inter-converted into erucin upon ingestion. We isolated erucin from Eruca sativa (Mill.) Thell. evaluated its hepatoprotective role in DMBA induced toxicity in male wistar rats. The rats were subjected to hepatic damage by five day regular intraperitoneal doses of DMBA. At the end of the protocol, the rats were euthanized, their blood was collected and livers were processed. The liver homogenate was analyzed for phase I (NADPH-cytochrome P450 reductase, NADH-cytochrome b5 reductase, cytochrome P450, cytochrome P420 and cytochrome b5), phase II (DT diaphorase, glutathione-S-transferase and γ-glutamyl transpeptidase) and antioxidant enzymes (superoxide dismutase, catalase, guaiacol peroxidise, ascorbate peroxidise, glutathione reductase and lactate dehydrogenase). The level of thiobarbituric acid reactive substances, lipid hydroperoxides, conjugated dienes and reduced glutathione in the liver homogenate was also analyzed. The serum was also analyzed for markers indicating hepatic damage (alkaline phosphatase, serum glutamic oxaloacetic transaminase, serum glutamic pyruvic transaminase, direct bilirubin and total bilirubin). Erucin provided significant protection against DMBA induced damage by modulating the phase I, II and antioxidant enzymes. The histological evaluation of liver tissue was also conducted, which showed the hepatoprotective role of erucin.

Conventional and modified hydrodistillation method for the extraction of glucosinolate hydrolytic products: a comparative account

Eruca sativa is extensively used as raw and its oil is also used for cooking due to its exceptional flavour. The volatile nature of the hydrolytic products of glucosinolates makes the extraction difficult. The hydrodistillation method used previously yield very less amount of the extract as well as the absence of stirring in the round bottom flask causes burning of both the crushed seeds and the flask. To overcome these drawbacks, a method has been developed using magnetic stirrer and hot plate. The yield and composition of hydrolytic products in the extract with the modified method was increased along with an increase in the amount of major hydrolytic products as seen by GC–MS. This method thus has immense potential in pharmaceutical industries, due to the ease of extraction and isolation.

Effect of cobalt(II) chloride hexahydrate on some human cancer cell lines

The present study investigates the anti-proliferative and apoptosis inducing mechanism of CoCl2·6H2O in PC-3 cancer cell line. Preliminary, three different forms of cobalt i.e., cobaltous (CoCl2·6H2O), macro-Co(II,III) oxide and nano-Co(II,III) oxide were screened for antiproliferative activity in PC-3 cell line. The CoCl2·6H2O being the most effective antiproliferative agent, hence it was further tested against lung (A549), prostrate (PC-3) and brain (IMR-32) cell lines. Human embryonic kidney cell line (293T) was used as a normal cell line to compare the toxicity of CoCl2·6H2O. The CoCl2·6H2O induced morphological and anatomical changes in PC-3 cancer cell which were studied using light, confocal and scanning electron microscopy. The lactate dehydrogenase was estimated which showed mild necrotic mode of cell death. The Annexin/PI staining confirmed the apoptotic mode of cell death in PC-3 cells. Further, production of reaction of reactive oxygen species and changes in mitochondrial membrane potential was also assessed spectrofluorimetrically. The cell cycle arrest was also investigated using flow cytometery. Finally, the caspase activity was estimated in CoCl2·6H2O treated PC-3 cancer cell line. Interestingly, it was found that CoCl2·6H2O induces more cell death in cancerous cells as compared to normal non-cancerous cells. These findings presented CoCl2·6H2O as potential antiproliferative agent.

β-Ionone derived apoptosis inducing endoperoxides; Discovery of potent leads for anticancer agents

A series of endoperoxides (3a-j) were synthesized and evaluated for cytotoxic activity against four human cancer cell lines by using SRB dye assay. All the compounds displayed moderate to high cytotoxic effect against almost all investigational cancer cells. Particularly, compounds bearing electron withdrawing groups such as nitro substituted compound 3j (IC50 = 0.001 μM) and fluoro substituted compound 3i (IC50 = 0.003 μM) showed comparatively more cytotoxic potential than standard drugs against lung cancer cell line (A549). All synthesized endoperoxides (3a-j) were further evaluated for their apoptotic potential through various parameters such as flow-cytometric analysis of nuclear DNA, flow-cytometric determination of mitochondrial membrane potential (ΔΨm), spectrofluorimetric estimation of intracellular ROS level and caspase-3 & 9 assays in treated lung cancer cells (A549); results reveal that endoperoxides induce apoptosis in cancer cells via mitochondrial pathway.

Synergistic antimutagenic effect of isothiocyanates against varied mutagens

Although plant foods provide an array of nutrients in the human diet, our knowledge of how these nutrients are interacting among each other at molecular level is limited. Among these, glucosinolates are the most important secondary metabolites, which are readily hydrolyzed to a variety of products including isothiocyanates. The current study emphasizes on the antimutagenic potential of three isothiocyanates (ITCs) viz. allyl, benzyl and 3-butenyl ITCs (individually and in binary combinations) in Ames Salmonella histidine reversion assay using Salmonella typhimurium TA98 and TA100 tester strains. Three different mutagens were used in the study including two direct acting mutagens [4-nitro-o-phenylenediamine (frame shift mutagen) and sodium azide (base substitution mutagen)] and one indirect acting mutagen [2-aminofluorene]. Furthermore, S9 fraction from rat liver microsomes was used to mimic the mammalian system. The study points towards the ability of ITCs to act as potent inhibitors of mutagenicity induced by different mutagens. Benzyl ITC was found to be the most effective among the three. Computation of combination effects was done on the basis of combination index values. It was observed that ITCs in combination induced a stronger antimutagenic response and that too at relatively low concentrations, without any signs of toxicity.

Alternative treatment strategies for neuropathic pain: Role of Indian medicinal plants and compounds of plant origin-A review

Neuropathic pain is a complex, chronic pain state accompanied by tissue injury and nerve damage. This important health issue constitutes a challenge for the modern medicine worldwide. The management of neuropathic pain remains a major clinical challenge, pertaining to an inadequate understanding of pathophysiological mechanisms of neuropathic pain. Various classes of drugs have been reported effective for the management of neuropathic pain viz. opiates, tricyclic antidepressants, and antiepileptic agents. However, association of adverse effects with these drugs hinders their confident prescription in people with neuropathic pain. Recently, various medicinal plants have been reported effective for the management of neuropathic pain. So, it may be prudent to look beyond synthetic drugs pertaining to their unprecedented pharmacotherapeutic effects with lesser adverse effects. The extensive literature review has been carried out from databases such as Science direct, Scifinder, Wiley online library, PubMed, Research gate, Google scholar and Chemical Abstracts. The list of Traditional Indian Medicinal plants (TIMPs) and isolated compounds have been compiled which have been reported effective as an alternative therapy for the management of neuropathic pain. This helps the researchers to discover some novel therapeutic agents against neuropathic pain.

Immunopotentiating significance of conventionally used plant adaptogens as modulators in biochemical and molecular signalling pathways in cell mediated processes

Natural products are of great surge in the identification of chemopreventive agents and biologically active molecules for the development of new promising therapeutic agents. These agents influence the cascade of biochemical and molecular signalling pathways involved in numerous physiological and pathological processes. The natural agents combat the dogma associated with the most dreaded, unconquered health concern and a multigenic disease- cancer. A category of plants known as adaptogens maintain perturbed homoeostasis, augment adaptations to noxious stimuli (exposure to cold, heat, pain, general stress, infectious organisms) and offer endurance to attenuate several disorders in human beings. The well known adaptogens and immunomodulators such as Rhodiola rosea, Withania somnifera, Tinospora cordifolia, Bacopa monnieri, Emblica officinalis, Glycyrrhiza glabra, Asparagus racemosus, Ocimum sanctum and Panax notoginseng claimed to have significant antioxidant and anticarcinogenic properties due to the presence of various biologically active chemical compounds. Their immunopotentiating activity is mediated through the modulation of T-cell immunity biochemical factors, transcription factors, some genes and factors associated with tumor development and progression. The combinatory formulation of active immunostimulating constituents from these plants may provide better homeostasis. These immunostimulant factors suggest their potential therapeutic significance in adjuvant or supportive therapy in cancer treatment.

Ameliorative potential of Alstonia scholaris (Linn.) R. Br. against chronic constriction injury-induced neuropathic pain in rats

BackgroundAlstonia scholaris commonly known as ‘Saptaparni’ is an Indian traditional medicinal plant used in Ayurveda. It is commonly used to treat various disorders like asthma, bronchitis, diarrhea, dysentery and malaria. In folklore medicine the milky juice of the plant is applied on wounds and ulcers to treat pain, ear ache and also in rheumatic pains.AimThe present study was designed to investigate the potential of A. scholaris R. Br. in chronic constriction injury of sciatic nerve (CCI) induced neuropathic pain in rats.MethodsPeripheral neuropathy was induced by chronic constriction injury of sciatic nerve. The behavioral parameters like mechanical and thermal hyperalgesia and cold allodynia were assessed on the 14th day. Tissue parameters like total protein, thiobarbituric acid reactive substances, reduced glutathione, myeloperoxidase, total calcium and TNF-α were assessed to check biochemical changes. Chloroform and methanol extract of A. scholaris leaves (100 and 200xa0mg/kg) and pregabalin (10xa0mg/kg, as positive control) were administered orally for 14 consecutive days starting from the day of surgery.ResultsCCI resulted in significant development of mechanical hyperalgesia, heat hyperalgesia and cold allodynia along with alteration in the biochemical changes. Administration of methanol extract at 200xa0mg/kg significantly attenuated the CCI induced change in nociceptive threshold and biochemical changes which was comparable to that of pregabalin. The high-performance liquid chromatography (HPLC) of the bioactive methanol extract revealed the presence of different types of flavonoids such as gallic acid, catechin, epicatechin, ellagic acid and kaempferol, in which kaempferol was observed to be in higher concentration.ConclusionMethanol extract (200xa0mg/kg) of A. scholaris showed the ameliorative effect in CCI induced neuropathic pain which may be due to the presence of kaempferol and attributed to its anti-oxidative and anti-inflammatory properties.

Development of validated high-temperature reverse-phase UHPLC-PDA analytical method for simultaneous analysis of five natural isothiocyanates in cruciferous vegetables

In the present study reverse-phase UHPLC-PDA technique was developed at 60°C for simultaneous quantification of allyl, 3-butenyl, 4-(methylthio)butyl, benzyl and phenethyl isothiocyanates. The validation parameter showed a very good linearity, with a correlation coefficient of 1.00 for all detected standard analytes. Also, high precision and accuracy were observed with lowest obtained values of 1.39% and 99.1%, respectively. Different varieties of three plants, viz. Brassica rapa var. rapa L., Raphanus sativus L. var. oleiformis Pers. and Eruca sativa Mill., were analyzed with this method. After analysis, 4-(methylthio)butyl isothiocyanate was observed to be the major component in the varieties of arugula. Allyl, benzyl and phenethyl isothiocyanates were detected in turnip varieties and, in addition, 3-butenyl isothiocyanate was detected in radish varieties.