Pankaj Pawar

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RESEARCH ARTICLE Open Access Evaluation of free-radical quenching properties of standard Ayurvedic formulation

Background: Cellular damage induced by free-radicals like Reactive Oxygen and Nitrogen Species (R... more Background: Cellular damage induced by free-radicals like Reactive Oxygen and Nitrogen Species (ROS and RNS) has been implicated in several disorders and diseases, including ageing. Hence naturally occurring anti-oxidant rich-herbs play a vital role in combating these conditions. The present study was carried out to investigate the in vitro free-radical quenching capacity of a known Ayurvedic poly-herbal formulation called Vayasthapana Rasayana. Methods: Methanol extracts of Vayasthapana Rasayana formulation (VRF) were studied for in vitro total antioxidant activity along with phenolic content and reducing power. In vitro assays like DPPH, FRAP, ABTS scavenging to evaluate radical quenching potential were performed. Results: The formulation has shown 94 % at 0.1 mg/ml DPPH free-radical scavenging activity as against 84 % at 0.1 mg/ml for standard ascorbic acid (IC50 value 5.51 μg/ml for VRF and 39 μg/ml for standard). It has a significant higher ferric reducing potential also (OD 0....

activity in TNBS-induced Inflammatory Bowel Disease

Background: Inflammatory Bowel Disease (IBD) is marked with chronic inflammation of intestinal ep... more Background: Inflammatory Bowel Disease (IBD) is marked with chronic inflammation of intestinal epithelium driven by oxidative stress. Traditional treatments with plant extracts gained renewed interest due to their ability to ameliorate the multi factorial conditions like inflammation. We investigated the beneficial effects of Withania somnifera in Trinitro Benzyl Sulfonic Acid (TNBS) induced experimental IBD through a rectally applicable formulation. Methods: The study included (i) preparation of gel formulation from aqueous Withania somnifera root extract (WSRE), (ii) biochemical assays to determine its performance potential, (iii) testing of formulation efficacy in TNBSinduced IBD rat model, and (iv) histo-patholgical studies to assess its healing and muco-regenerative effect in IBDinduced rats. For this purpose, concentration dependant antioxidant activity of the extracts were evaluated using biochemical assays like (a) inhibition of lipid peroxidation, (b) NO scavenging, (c) H2O...

Characterization of a Bowman–Birk type trypsin inhibitor purified from seeds of Solanum surattense

Scientific Reports

A Bowman–Birk type trypsin inhibitor protein (SSTI) from seeds of the medicinal plant Solanum sur... more A Bowman–Birk type trypsin inhibitor protein (SSTI) from seeds of the medicinal plant Solanum surattense was isolated, purified and characterized. SSTI showed a single band on SDS-PAGE corresponding to 11.4 kDa molecular weight. It is a glycoprotein (2.8% glycosylation) that differentially interacted with trypsin and chymotrypsin in a concentration-dependent manner. Its peptide sequence is similar to other Bowman–Birk type protease inhibitors found in Glycine max and Phaseolus acutifolius. The inhibitory activity was stable over a wide range of pH (1–10) and temperatures (10–100° C). Far-UV Circular Dichroism (CD) studies showed that SSTI contains β sheets (~ 23%) and α helix (~ 6%) and demonstrated structural stability at wide pH and high temperature. The kinetic analysis revealed a noncompetitive (mixed) type nature of SSTI and low inhibitor constant (Ki) values (16.6 × 10−8 M) suggested strong inhibitory activity. Isothermal titration calorimetric analysis revealed its high affin...

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A Technique for Rapid Micropropagation of Solanum surattense Burm. f

In Solanulll SllraUense, a medicinally important member of family Solanaceae, rapid micropropagat... more In Solanulll SllraUense, a medicinally important member of family Solanaceae, rapid micropropagation and di rect organogenesis was achieved from shoot tip and leaf explants, respectively. Both leaf and shoot tips differentiated into number of small shoots without apparently forming any callus. Average IS0-200 adventitious shoots were pro duced from a single explant on MS basal medium fortified with BAP and kinetin individually and in combination. Caulogenesis was followed by rhizogenesis on the same medium without phytohormone. III vitro regenerated plantlets were healthy with dark green leaves and attained a length of 5-10 cm in 10 weeks. The post- transplantation survival rate in the field was SO±5% without any seasonal constraints.

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Isolation and characterization of solasodine from cultured cells of Solanum khasianum

A Protein α-Amylase Inhibitor from Withania Somnifera and its Role in Overall Quality and Nutritional Value Improvement of Potato Chips during Processing

Food and Bioprocess Technology

Cold storage and processing increase the reducing sugar level in potato (Solanum tuberosum L.) wh... more Cold storage and processing increase the reducing sugar level in potato (Solanum tuberosum L.) which is responsible for browning and acrylamide formation that adversely affect sensory and nutrient quality of chips. Effect of α-amylase inhibitor from Withania somnifera (WSAI) treatment on the overall quality improvement of potato chips during processing was studied. WSAI treatment to potato slices at 200 ppm for 30 min was found to reduce browning (60%), residual amylase, and polyphenol oxidase activities (~ 40%) and reduce sugar level by 25%, respectively, over control. Color match analysis indicated an improvement in whiteness and brightness indices and a significant reduction in yellowness index of potato chips. The treatment proved to be superior over blanching and reduction in acrylamide generation during frying was which also observed in chips treated with it. Furthermore, our results were comparable to that of treatment with α-amylase inhibitor of Triticum aestivum and better than synthetic inhibitor, acarbose.

Genomic and functional characterization of coleopteran insect-specific α-amylase inhibitor gene from Amaranthus species

Plant molecular biology, 2017

The smallest 32 amino acid α-amylase inhibitor from Amaranthus hypochondriacus (AAI) is reported.... more The smallest 32 amino acid α-amylase inhibitor from Amaranthus hypochondriacus (AAI) is reported. The complete gene of pre-protein (AhAI) encoding a 26 amino acid (aa) signal peptide followed by the 43 aa region and the previously identified 32 aa peptide was cloned successfully. Three cysteine residues and one disulfide bond conserved within known α-amylase inhibitors were present in AhAI. Identical genomic and open reading frame was found to be present in close relatives of A. hypochondriacus namely Amaranthus paniculatus, Achyranthes aspera and Celosia argentea. Interestingly, the 3'UTR of AhAI varied in these species. The highest expression of AhAI was observed in A. hypochondriacus inflorescence; however, it was not detected in the seed. We hypothesized that the inhibitor expressed in leaves and inflorescence might be transported to the seeds. Sub-cellular localization studies clearly indicated the involvement of AhAI signal peptide in extracellular secretion. Full length r...

A glycoprotein α-amylase inhibitor from Withania somnifera differentially inhibits various α-amylases and affects growth and development of Tribolium castaneum

Pest Management Science, 2016

Identification and characterisation of plant defensive molecules enrich our resources to design c... more Identification and characterisation of plant defensive molecules enrich our resources to design crop protection strategies. In particular, plant-derived proteinaceous inhibitor(s) of insect digestive enzymes appear to be a safe, sustainable and attractive option. A glycoprotein having non-competitive α-amylase inhibitory activity with a molecular weight of 8.3 kDa was isolated and purified from seeds of Withania somnifera α-amylase inhibitor (WSAI). Its mass spectrometry analysis revealed 59% sequence coverage with Wrightide II-type α-amylase inhibitor from Wrightia religiosa. A dose-dependent inhibition of α-amylases from Aspergillus oryzae, Bacillus subtilis, Helicoverpa armigera and Tribolium castaneum was recorded. Interestingly, WSAI did not inhibit human salivary α-amylase significantly. When adults of T. castaneum were fed with WSAI (1.6 mg g(-1) ), decrease in consumption, growth and efficiency of conversion of ingested food was evident, along with over fourfold increases in feeding deterrence index. A decline in larval residual α-amylase activity after feeding of WSAI resulted in a reduction in longevity of T. castaneum. The study reflects the significance of WSAI in affecting the overall growth and development of T. castaneum. Pre- and post-harvest pest resistive capability makes WSAI a potential candidate for insect pest management. Further, the effectiveness of this inhibitor could be explored either in formulations or through a transgenic approach. © 2016 Society of Chemical Industry.

Characterization of two coleopteran α-amylases and molecular insights into their differential inhibition by synthetic α-amylase inhibitor, acarbose

Insect biochemistry and molecular biology, Jan 27, 2016

Post-harvest insect infestation of stored grains makes them unfit for human consumption and leads... more Post-harvest insect infestation of stored grains makes them unfit for human consumption and leads to severe economic loss. Here, we report functional and structural characterization of two coleopteran α-amylases viz. Callosobruchus chinensis α-amylase (CcAmy) and Tribolium castaneum α-amylase (TcAmy) along with their interactions with proteinaceous and non-proteinaceous α-amylase inhibitors. Secondary structural alignment of CcAmy and TcAmy with other coleopteran α-amylases revealed conserved motifs, active sites, di-sulfide bonds and two point mutations at spatially conserved substrate or inhibitor-binding sites. Homology modeling and molecular docking showed structural differences between these two enzymes. Both the enzymes had similar optimum pH values but differed in their optimum temperature. Overall, pattern of enzyme stabilities were similar under various temperature and pH conditions. Further, CcAmy and TcAmy differed in their substrate affinity and catalytic efficiency towa...

Isolation and survey of antibiotic sensitivity in nosocomial infections in north Maharashtra region

The Journal of the Association of Physicians of India, Jul 1, 2013

Background: The present work was aimed to study the prevalence of the drug resistance among Gram ... more Background: The present work was aimed to study the prevalence of the drug resistance among Gram positive and Gram negative organisms isolated from clinical samples obtained from various private nursing homes of Jalgaon city and surrounding areas in the state of Maharashtra. Materials and methods: Different pathogenic organisms were isolated from one thousand seven hundred and ninety clinical samples obtained from various nursing homes of Jalgaon region of North Maharashtra, MS, India during the period of August, 2010 to July, 2011. Antibiotic sensitivity testing of isolated pathogenic organisms was done with fifty different antibiotics of 13 different groups by the disk diffusion technique using Mueller Hinton Agar Plates according to CLSI (formerly NCCLS) guidelines. Results: Out of total samples analyzed 1171 samples showed the presence of infection while in 619 samples no growth of organisms was seen on culture medium. Microbial growth revealed presence of 11 different organisms (E. coli, Klebsiella spp., Proteus spp., Pseudomonas spp., Enterobacter spp., Salmonella spp., Acinetobater spp., Streptococccus spp., Staphylococcus aureus (MRSA and MSSA)) including Candida spp. Sensitivity profile of these organisms towards different antibiotics is reported here. Conclusion: The study resulted in generation of representative status of the antibiotic sensitivity pattern of commonly found organisms in nosocomial infections in the region. This could be useful for the clinicians to choose correct antibiotic and ensure the judicious use of the same for their patients.

Isolation and survey of antibiotic sensitivity in nosocomial infections in north Maharashtra region

The Journal of the Association of Physicians of India, 2013

The present work was aimed to study the prevalence of the drug resistance among Gram positive and... more The present work was aimed to study the prevalence of the drug resistance among Gram positive and Gram negative organisms isolated from clinical samples obtained from various private nursing homes of Jalgaon city and surrounding areas in the state of Maharashtra. Different pathogenic organisms were isolated from one thousand seven hundred and ninety clinical samples obtained from various nursing homes of Jalgaon region of North Maharashtra, MS, India during the period of August, 2010 to July, 2011. Antibiotic sensitivity testing of isolated pathogenic organisms was done with fifty different antibiotics of 13 different groups by the disk diffusion technique using Mueller Hinton Agar Plates according to CLSI (formerly NCCLS) guidelines. Out of total samples analyzed 1171 samples showed the presence of infection while in 619 samples no growth of organisms was seen on culture medium. Microbial growth revealed presence of 11 different organisms (E. coli, Klebsiella spp., Proteus spp., Ps...

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Compendium of Transgenic Crop Plants, 2009

Evaluation of comparative free-radical quenching potential of Brahmi (Bacopa monnieri) and Mandookparni (Centella asiatica)

AYU (An International Quarterly Journal of Research in Ayurveda), 2011

Ayurvedic texts describe rejuvenate measures called Rasayana to impart biological sustenance to b... more Ayurvedic texts describe rejuvenate measures called Rasayana to impart biological sustenance to bodily tissues. Rasayana acting specifi cally on brain are called Medhya Rasayana. Brahmi is one of the most commonly practiced herbs for the same. Yet there exist a controversy regarding the exact plant species among Bacopa monnieri L. Penn (BM) and Centella asiatica (L.) Urban (CA) to be used as Brahmi in the formulations. Though the current literature available has suggested a very good nootropic potential of both the drugs, none of the studies have been carried out on comparative potential of these herbs to resolve the controversy. Free-radical scavenging potential for these plants is studied to fi nd out their comparative effi cacy. The study revealed a very good in vitro free-radical scavenging properties of aqueous and ethanolic extracts of both the plants as evidenced by FRAP, DPPH, reducing power, and antilipid peroxidation assays. It can be concluded from the studies that both the plants, although taxonomically totally different at family level, showed similar type of in vitro activities. The total phenolic and fl avonoid contents also revealed a signifi cant similarity in the two plants. The in vitro study supports the Ayurvedic concept of BM and CA having a similar potential.

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Assessment of Nutritional Value of Some Wild Edible Plants from Satpura Hills of Maharashtra, India

... Phytochemical screening of the plants indicated that almost all plants are important fromneut... more

Traditional ethnomedicinal knowledge confined to the Pawra tribe of Satpura hills, Maharashtra, India

Ethnobotanical surveys can potentially bring out many different clues for the development of safe... more Ethnobotanical surveys can potentially bring out many different clues for the development of safe, effective and inexpensive indigenous remedies. Present study has been focussed on Pawra tribe of Nandurbar district of Maharashtra, India. Field surveys of this area were carried out during 2000-2005 through several field visits and interviews. The aim of the present study was primarily to evaluate and inverntorize medicinal uses of the plants known to Pawras and to encourage preservation of their culture, conservation and sustainable utilization of the plant wealth. After an extensive comparison with literature, about 80 unique or less known uses of 79 plant species belonging to 59 families are described. The documented ethnomedicinal usage of plants mostly pertains to cure asthma, inflammation, lactation, menstrual problems, poisonous bites, skin problems, stomach ache and tooth ache. We found that the study area is rich in ethnic and biodiversity and the tribe posses a valuable treasure of ethnobotanical knowledge. This wisdom available with the tribe is transmitted only through oral communication in locally-spoken, script-less language and therefore needs conservation. The plants used by the Pawra tribe are arranged alphabetically followed by family name, herbarium number, local name, parts used, mode of preparation and medicinal uses.

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Agrobacterium rhizogenes mediated hairy root induction in two medicinally important members of family Solanaceae

Withania somnifera (L) Dunal. and Solanum surattense Burm f., the two medicinally important membe... more Withania somnifera (L) Dunal. and Solanum surattense Burm f., the two medicinally important members of family Solanaceae, were investigated for induction of hairy roots using soil borne bacterium, Agrobacterium rhizogenes. Explants like stem segments, hypocotyls and leaves with midrib were infected with the bacterium in vitro. In both plants, extensive hairy roots were induced from leaf explant containing midrib within 15 days of infection. These roots were then established on MS basal medium and their growth was observed to be independent of exogenous supply of phytohormones. The growth rate of transformed roots was 10-fold as compared to control. These roots can be grown and established in phytohormone free liquid MS medium and used as a promising source of secondary metabolites of medicinal significance.

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Demonstration of the potential of Hibiscus cannabinus Linn. flowers to manage oxidative stress, bone related disorders and free-radical induced DNA damage

Indian Journal of Natural Products and Resources, 2010

Hibiscus cannabinus Linn. flowers (HCF) were investigated for free-radical scavenging properties ... more Hibiscus cannabinus Linn. flowers (HCF) were investigated for free-radical scavenging properties in vitro, capacity to protect DNA from oxidative damage and inhibiting gelatinolytic activity of collagenase type I and II. For this purpose, flowers extracted with methanol and water and subjected to DPPH free radical scavenging assay, calculation of IC 50 values, using vitamin C as a standard, reducing power assay, DNA damage protection assay and inhibition of gelatinolytic activity of collagenase type I and II. The DPPH free radical scavenging activity ranged from 440 to 700μg/ml for different extracts vis-à-vis 39 μg/ml for standard vitamin C. A similar trend was visible in reducing power activity. Both activities reflected a strong anti-oxidant potential of HCF and in turn against stress. Further, both extracts at 100 μg/ml (i) were efficient in protecting DNA against oxidative damage and (ii) exhibited inhibition of gelatinolytic activity of collagenase type I up to 87% and type II...

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Phytoremediation of fluoride with garden ornamentals Nerium oleander, Portulaca oleracea, and Pogonatherum crinitum

Environmental science and pollution research international, 2017

Nursery grown plants of Nerium oleander, Pogonatherum crinitum, and Portulaca oleracea were obser... more Nursery grown plants of Nerium oleander, Pogonatherum crinitum, and Portulaca oleracea were observed to remove fluoride up to 92, 80, and 73%, respectively, from NaF solution at the concentration of 10 mg L(-1) within 15 days. Concentration range of 10-50 mg L(-1) of fluoride revealed a constant decrease of removal from 92 to 51% within 15 days by N. oleander, while the biomass (one to five plants) showed enhancement in removal from 74 to 98% in 10 days. Translocation and bioaccumulation factors calculated after fluoride contents in roots and leaves of N. oleander, P. crinitum, and P. oleracea were 1.85, 1.19, and 1.43, and 9.8, 3.6, and 2.2, respectively. P . oleracea, P. crinitum, and N. oleander showed reductions in chlorophyll contents by 40, 57 and 25 and 8%, carbohydrates by 50, 44, and 16%, and proteins by 38, 53, and 15%, respectively. Activities of superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPX) in the roots of P. oleracea, P. crinitum, and N. o...

Evaluation of free-radical quenching properties of standard Ayurvedic formulation Vayasthapana Rasayana

BMC Complementary and Alternative Medicine, 2011

Background Cellular damage induced by free-radicals like Reactive Oxygen and Nitrogen Species (RO... more Background Cellular damage induced by free-radicals like Reactive Oxygen and Nitrogen Species (ROS and RNS) has been implicated in several disorders and diseases, including ageing. Hence naturally occurring anti-oxidant rich-herbs play a vital role in combating these conditions. The present study was carried out to investigate the in vitro free-radical quenching capacity of a known Ayurvedic poly-herbal formulation called Vayasthapana Rasayana. Methods Methanol extracts of Vayasthapana Rasayana formulation (VRF) were studied for in vitro total antioxidant activity along with phenolic content and reducing power. In vitro assays like DPPH, FRAP, ABTS scavenging to evaluate radical quenching potential were performed. Results The formulation has shown 94% at 0.1 mg/ml DPPH free-radical scavenging activity as against 84% at 0.1 mg/ml for standard ascorbic acid (IC50 value 5.51 μg/ml for VRF and 39 μg/ml for standard). It has a significant higher ferric reducing potential also (OD 0.87 at...

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