Pallavi Thakur

Evidence based herbal drug standardization approach in coping with challenges of holistic management of diabetes: a dreadful lifestyle disorder of 21st century

Plants by virtue of its composition of containing multiple constituents developed during its growth under various environmental stresses providing a plethora of chemical families with medicinal utility. Researchers are exploring this wealth and trying to decode its utility for enhancing health standards of human beings. Diabetes is dreadful lifestyle disorder of 21st century caused due to lack of insulin production or insulin physiological unresponsiveness. The chronic impact of untreated diabetes significantly affects vital organs. The allopathic medicines have five classes of drugs, or otherwise insulin in Type I diabetes, targeting insulin secretion, decreasing effect of glucagon, sensitization of receptors for enhanced glucose uptake etc. In addition, diet management, increased food fiber intake, Resistant Starch intake and routine exercise aid in managing such dangerous metabolic disorder. One of the key factors that limit commercial utility of herbal drugs is standardization. Standardization poses numerous challenges related to marker identification, active principle(s), lack of defined regulations, non-availability of universally acceptable technical standards for testing and implementation of quality control/safety standard (toxicological testing). The present study proposed an integrated herbal drug development & standardization model which is an amalgamation of Classical Approach of Ayurvedic Therapeutics, Reverse Pharmacological Approach based on Observational Therapeutics, Technical Standards for complete product cycle, Chemi-informatics, Herbal Qualitative Structure Activity Relationship and Pharmacophore modeling and, Post-Launch Market Analysis. Further studies are warranted to ensure that an effective herbal drug standardization methodology will be developed, backed by a regulatory standard guide the future research endeavors in more focused manner.

Attenuation of adhesion, quorum sensing and biofilm mediated virulence of carbapenem resistant Escherichia coli by selected natural plant products.

The multi-drug resistance offered by Carbapenem Resistant Escherichia coli (Family: Enterobacteriaceae; Class: Gammaproteobacteria) against third line antibiotics can be attributed towards its ability to develop biofilm. Such process involves adhesion and quorum-sensing induced colonization leading to biomass development. The present study explored the anti-adhesion, anti-quorum sensing and anti-biofilm potential of 05 pre-standardized potent herbals. Berberis aristata (PTRC-2111-A) exhibited maximum potential in all these activities i.e. 91.3% ± 0.05% (Anti-adhesion), 96.06% ± 0.05% (Anti-Quorum sensing) and 51.3% ± 0.07% (Anti-Biofilm formation) respectively. Camellia sinensis (PTRC-31911-A) showed both anti-adhesion (84.1% ± 0.03%) and anti-quorum sensing (90.0%) potential while Holarrhena antidysenterica (PTRC-8111-A) showed only anti-quorum sensing potential as compared to standards/antibiotics. These findings were in line with the molecular docking analysis of phytoligands against Lux S and Pilin receptors. Furthermore, the pairwise correlation analysis of the tested activities with qualitative, quantitative and bioactivity functional descriptors revealed that an increased content of alkaloid, moderate content of flavonoids and decreased content of tannins supported all the three activities. In addition, nitric oxide and superoxide scavenging activity were found to be correlated with anti-quorum sensing activity. The findings indicated clearly that B. aristata (Family: Berberidaceae) and C. sinensis (Family: Theaceae) were potent herbal leads with significant therapeutic potential which further needs to be explored at pre-clinical level in the future.

Camellia sinensis Ameliorates the Efficacy of Last Line Antibiotics Against Carbapenem Resistant Escherichia coli.

Aquo‐ethanolic extract of Camellia sinensis (PTRC‐31911‐A), standardized using Fourier transform infrared analysis, was found to have seven common functional groups in comparison with pre‐identified marker compound ‘quercetin’. Phyto‐chemical quantitation analysis revealed the presence of 10.65 µg/mg of flavonoids. The bioactivity fingerprint profile of PTRC‐31911‐A includes IC50 (Hydroxyl radical site specific scavenging) = 11.36 ± 0.5 µg/mL, IC80 (Hydroxyl radical non‐site specific scavenging) = 26.44 ± 0.5 µg/mL and IC50 (Superoxide ion scavenging) = 10.141 ± 0.5 µg/mL. The drug combination analysis of PTRC‐31911‐A with five third‐line antibiotics was carried out against carbapenem‐resistant Escherichia coli. The analysis of combination of PTRC‐31911‐A (6.25–1000 µg/mL) and antibiotics (6.25–1000 µg/mL) revealed synergistic behaviour (fractional inhibitory concentration indices < 1) with tigecycline, ertapenem, meropenem, colistin and augmentin. The lead combination of PTRC‐31911‐A + ertapenem or meropenem showed maximum augmentative potential at 50 and 100 µg/mL, respectively, with nearly five‐fold decrease in minimum inhibitory concentrations as compared with respective antibiotics alone. The synergistic effects implied that the antibacterial combinations of PTRC‐31911‐A and ertapenem, meropenem, colistin, tigecycline or augmentin would be more effective than a single monotherapy with either of the antibacterial agent. Copyright

Targeting Antibiotic Resistant Salmonella enterica: Bio-matrix Based Selectionand Bioactivity Prediction of Potential Nutraceuticals

Aim: Potential herbal leads as novel therapeutic alternatives against Multi-Drug Resistant Salmonella enterica serovar typhimurium were investigated and validated by mol inspiration. Methodology: The present study utilizes an in silico ‘Herbal Informatics’ model which deploys dynamic search protocols, priority indexing and systemic categorization for rationale based selection of nutraceuticals targeting the critical virulence factors of Multi-Drug Resistant Salmonella enterica serovar typhimurium. Furthermore, in silico biochemical activity prediction was conducted using ‘Mol inspiration’ chemiinformatics tool, so as to propose the drug likeliness of selected natural plant products. Results: Out of the 05 selected bioactivity parameters of Salmonella enterica, lipo-polysaccharide inhibition exhibited maximum relevance as physiological target i.e., 65%, followed by other parameters like enterochelin inhibition, Type III secretory system inhibition, Superoxide dismutase inhibition and Symptomatic relief provision. The binary matrix analysis of database of 50 plants identified using classical bioprospection filtered 28 herbals which exhibited probable potential to mitigate 03 or more virulence factors. The weightage matrix analysis further scrutinized the selection upto 10 herbals having a score more than median weightage matrix score i.e., 14.98. The optimization of data on a scale of 0-1 using fuzzy score matrix analysis led to the final selection of 10 herbals i.e., Abrus precatorius, Azadirachta indica, Camellia sinensis, Holarrhena antidysenterica, Andrographis paniculata, Adhatoda vasica, Euphorbia hirta, Ocimum sanctum, Terminalia arjuna and Terminalia belerica. In silico Bioactivity prediction analysis of predominant phytoconstituents of selected herbals revealed Holarrhena antidysenterica (Conessine), Andrographis paniculata (Andrographolide), Euphorbia hirta (Amyrin) and Terminalia arjuna (Arjunolic acid) exhibiting drug likeliness with their targeted action as a GPCR Ligand, Nuclear Receptor Ligand or Protease inhibitor.

Augmenting the potency of third-line antibiotics with Berberis aristata: In vitro synergistic activity against carbapenem-resistant Escherichia coli

The aim of this study was to analyse the in vitro synergistic antibacterial potential of an aquoethanolic extract of the stem bark of Berberis aristata (PTRC-2111-A) with third-line antibiotics against carbapenem-resistant Escherichia coli. PTRC-2111-A was prepared and was characterised using phytochemical- and bioactivity-based fingerprinting. Fourier transform infrared spectroscopy (FTIR) and liquid chromatography-mass spectrometry (LC-MS) analyses were performed, and superoxide and hydroxyl scavenging activities were assessed in conjunction with in vitro antimicrobial efficacy testing against the test micro-organism. Analysis of drug combinations of PTRC-2111-A and third-line antibiotics was performed using CompuSyn software. PTRC-2111-A from B. aristata was found to have seven common functional groups in comparison with the pre-identified marker compound quercetin, and phytochemical quantitation analysis revealed the presence of 25.44% alkaloids. Moreover, PTRC-2111-A was found to contain isoquinoline alkaloids, namely berbamine, berberine, reticuline, jatrorrhizine, palmatine and piperazine, as elucidated in the LC-MS analysis. Analysis of combinations of PTRC -2111-A and antibiotics revealed synergistic behaviour [fractional inhibitory concentration index (FICI)<1] with colistin, tigecycline and amoxicillin/clavulanate potassium (Augmentin(®)), whereas antagonism (FICI>1) was seen with ertapenem and meropenem.

Protective effect of Berberis aristata against peritonitis induced by carbapenem-resistant Escherichia coli in a mammalian model

OBJECTIVES Berberis aristata is known to contain a variety of phenolic compounds contributing to its holistic capability of mitigating bacterial multidrug resistance. METHODS B. aristata stem bark extract was prepared and was characterised using liquid chromatography-mass spectrometry (LC-MS). The antimicrobial efficacy of the extract against carbapenem-resistant Escherichia coli was assessed in vivo in an animal model using Sprague Dawley rats. Microbial counts in blood and urine, physical health status, haematological and biochemical analysis of blood, and histopathology of the kidney were assessed as the study endpoints. RESULTS An aquo-alcoholic extract of B. aristata (PTRC-2111-A) was found to effectively manage peritonitis induced by carbapenem-resistant E. coli in a rat model at a single post-exposure prophylactic dose of 0.5mg/kg body weight (BW). The extract was also found to show a no observed adverse effect level (NOAEL) up to a dose of 2000mg/kg BW. Physical, immunological, haematological, biochemical and histopathological aberrations were found to be restored to normal in the herbal-treated group at a dose of 0.5mg/kg BW. CONCLUSIONS The antimicrobial and hepatorenal protective ability of PTRC-2111-A could be attributed to the presence of isoquinoline alkaloids.

Assessment of aquo-ethanolic extract of Camellia sinensis against Carbapenem Resistant Escherichia coli: In Vivo Trials in a Murine Model.

The prevalence of Carbapenem Resistant Escherichia coli (CRE) has increased considerably during the last decade, which can be ascribed to relative scarcity of effective non toxic antimicrobial agents. The present study was conducted to evaluate the antimicrobial activity of aquo-ethanolic (1:1) extract of leaves of Camellia sinensis (PTRC-31911-A) against Carbapenem Resistant Escherichia coli at preclinical level using peritonitis infection model in Sprague Dawley rats. Efficacy analysis of PTRC-31911-A involved enumeration of CRE colonies in blood and urine samples of test animals for a period of 5 days from infection. A reduction in microbial count of biological fluids was considered as the primary endpoint of the selected murine model. Physical, biochemical, hematological and histological indices of toxicity were employed as secondary relative indicators of the induced disease. Physical manifestations of infected rats included significantly high body temperature (TempInfected=103.18°F, ∼5% increase) and noteworthy reduction in weight (WeightInfected=126.83g, ∼15% decrease) as compared to control. Significant (P<0.05) increase in total white blood cells, eosinophil and monocyte counts as well as a significant decrease (P<0.05) in erythrocytes count, hematocrit volume, red blood cell distribution width and hemoglobin concentration were observed in the infected group as compared to the control group. Furthermore, noteworthy increase in liver and kidney function test parameters were observed in case of infected groups. All the hematological and biochemical parameters were found to be within optimum range in case of treatment group, indicating restoration of homeostasis. Histopathological studies also presented symptoms of hemorrhage and glomerular damage with structural distortion in glomerular capillary loops of infected groups, which were later recovered in treated groups, indicating the nephro-protective potential of PTRC-31911-A. The study clearly points out that Camellia sinensis extract (PTRC-31911-A; single dose of 5mg/Kg bwt; oral,+24h) is highly effective against Carbapenem Resistant Escherichia coli owing mainly to the presence of flavonoids and polyphenolic compounds, identified by LCMS. Ongoing studies are expected to further unravel the mechanism of action and bioactivity determinants of this broad spectrum plant extract.

In vitro bactericidal activity of Berberis aristata extract against clinical isolates of carbapenem-resistant Escherichia coli.

Abstract Background: Berberis aristata is known to contain a variety of phenolic compounds, flavonoids such as quercetin attributing towards its holistic capability of mitigating multidrug resistance. Methods: B. aristata stem bark extract was prepared and characterized using phytochemical and bioactivity-based fingerprinting. Anti-oxidant and anti-lipid peroxidation profiling was also done in conjunction with in vitro anti-microbial efficacy testing against the test microorganism i. e., New Delhi Metallo-β-lactamase-1 (NDM-1) Escherichia coli. Results: Aquo-alcoholic (1:1) extract of B. aristata (PTRC-2111-A), containing 3.0±0.02 µg of QUERCETIN/mg of dried extract, exhibited [flavonoid/polyphenol: F/P (quercetin %) ~ 0.16(0.06 %)]. The bioactivity fingerprint profile of PTRC-2111-A included IC50 ratio [DPPH/NOS]=0.064 as functional standardized value having IC50 (DPPH Scavenging)=16±0.5 µg/mL and IC50 (Nitric Oxide Scavenging)=250±0.5 µg/mL respectively. The reducing ability and anti-lipid peroxidation equivalent (extract: standard) of PTRC-2111-A with respect to standard was estimated to be 3.44 (ascorbic acid) and 0.78 (quercetin) respectively. In vitro anti-microbial activity evaluated against sts-09 multidrug-resistant strain of carbapenem-resistant E. coli was found to be 25 µg/mL. Conclusions: B. aristata was found to contain a number of phytoconstituents, which acts in a synergistic manner to provide significant bactericidal potential against carbapenem-resistant E. coli.

Development of herbal based bioprotective agents against microorganisms of biological importance

Malnutrition is India’s silent emergency and among India’s greatest human development challenges. Although India has seen strong economic growth over the past 20 years, malnutrition in children under five years of age continues to be among the highest in the world. Rates of malnutrition among India’s children are almost five times more than in China, and twice those in Sub-Saharan Africa. Nearly half of all India’s children approximately 60 million are underweight, about 45% are stunted (too short for their age), 20% are wasted (too thin for their height, indicating acute malnutrition), 75% are anemic, and 57% are Vitamin A deficient. Malnutrition affects children’s chances of survival, increases their susceptibility to illness, reduces their ability to learn, increases their chances of dropping out early from school, and makes them less productive in later life. Much of this undernourishment happens during pregnancy and in the first two years of a child’s life and, without appropriate interventions, the damage to brain development and future economic productivity is largely irreversible. Can we stop this Silent Emergency of India? Biography Dipali Kendre, Sonali Fere, Rajnandani Ingale & Pooja Jadhav are Second year BHMS students from V.K. Homoeopathic Medical College & Hospital, Latur, under Maharashtra University of Health Sciences, Nashik.Aging is believed to be a first-order risk factor for most neurodegenerative disorders. Brain changes do not occur to the same extent in all brain regions.7 Men and women may also differ with frontal and temporal lobes most affected in men compared with the hippocampus and parietal lobes in women. The neurotransmitters most often discussed with regard to ageing are dopamine, serotonin and acetyl-choline. Vestibular stimulation modulates the neuro-transmitters which are involved in brain aging and delay aging. Hence we recommend controlled vestibular stimulation to all. This in the need of time to identify the importance of vestibular system and to start translational research in this area.H is a life-style disorder. Hypertension causes 6% of death worldwide. Data from prospective observational studies and from randomised control of antihypertensive agent have shown a consistent continuous linear relation between usual blood pressure level and risk of coronary heart disease and stroke. Hypertension is commonly taken as prolong and excess of tension and stress caused by blood on arteries. It is an elevated arterial pressure level >140/90 mm of Hg. Though few theories are proposed for the hypertension still the etiopathogeneis of hypertension is not crystal clear. Scatted references are available in ayurvedic literature indicative of etio-pathogenesis and complications of hypertension. Hypertension is Tridoshaja Vata-pitta pradhan disorder with Dusthi of Rasa, Rakta, meda. The causative factor for Rakta and Rasa Dusthi are excessive intake of salt, alcoholic beverages, fatty diet and worries, which are similar to the causes of hypertension. The clinical picture of Rakta Dusthi includes bhrama, Daha, Shirashool similar to that of clinical features of hypertension. In hypertension there is increased resistance to the flow of blood in arteries resulting into increase pressure over arterial wall this indicates towards the involvement of Vata, Raktavha srotasa and Rasa-Rakta Dusthi and Pitta. Arteriosclerosis is indicative of Dhamani pratichaya which is one of the Kaphaja Nanatmaja Vikara this indicates the involvement of Kapha and Medo Dhatu in Samprapti of hypertension. The details of understanding of etiopathogenesis, symptomatology, complication of hypertension and its line of treatment according to Ayurveda will be presented in full paper during seminar. Ujwala Hivale et al., Altern Integ Med 2013, 2:10 http://dx.doi.org/10.4172/2327-5162.S1.003F is an important factor for male and female in life. In our day to day life, the stress and strain are the causes of male infertility. Spermatogenesis was suppressed at all the stages of cell division and maturity due to stress. Many natural nontoxic herbs were found to enhance the fertility in male. This study involves the effect of the herbs, Cycas circinalis and Ionidium suffruticosum in improving the fertility of stress induced male albino rats. A total of 50 healthy young male albino rats (1012 weeks) were selected and are subjected to cold water swimming stress for 10 minutes, a day for a period of one month to check the stress induced status and serum corticosterone was analyzed. Cycas and Ionidium extract 200 mg/kg body weight were administered orally to the experimental albino rats E-1 (n=10) and E-2 (n=10) respectively and compared to the positive control albino rats C-1 (n=10) administered with testosterone 10 mg/kg body weight subcutaneously, biweekly with that of the stressed control rats C-2 (n=10) using various parameters such as sexual behaviour, weight of animals, dimension of testes, hormonal and semen analysis, histological analysis of testes and histomorphometry of testes. The drug’s efficacy was proved by the restitution of fertility by comparing with the normal fertile controls C-3 (n=10). The administration of the drug showed significant improvement in all the parameters in experimental rats when compared to control rats. The herb was found to be effective on the gonads of stress induced sterile male albino rats. B. Senthil Kumar et al., Altern Integ Med 2013, 2:10 http://dx.doi.org/10.4172/2327-5162.S1.003Based on clinical signs and symptoms psoriasis can be compared to ekakushta in Ayurveda. Ekakushta is one of the kshudra kushta (obstinate skin disease) characterized by unhydrosis, patches wide-spreaded with fishy silvery scales. Though several drugs are in practice for the treatment of psoriasis, still cure or better relief is awaited destiny on the other hand these medications are associated with severe side effects. Whereas Ayurveda emphasises treatment of disease in holistic approach particularly correcting the root cause through samshodhana (panchakarma), specifically for skin diseases like psoriasis, vamana (therapeutic emesis), virechana (therapeutic purgation) is advised. In clinical practice repeated administration of these therapies showed significant improvement. The present case was a male subject aged 19 yrs suffering with Guttate type of psoriasis since four years. To whom classical virechana karma was administered in a methodic way. This has shown improvement, scaling, discolouration, elevated patches and itching were reduced. Now, the patient is under observation with simple oral medication. The case study with treatment in detail will be presented during the seminar.Over Recent years advancement in nanoparticles drug delivery is widely expected to change the landscape of pharmaceutical industries for the foreseeable future. Nanotechnologies have become a significant priority worldwide. Several manufactured nanoparticles - particles with one dimension less than 100 nm – are increasingly used in consumer products. At nanosize range, the properties of materials differ substantially from bulk materials of the same composition, mostly due to the increased specific surface area and reactivity, which may lead to increased bioavailability and toxicity. Thus, for the assessment of sustainability of nanotechnologies, methods of manufacturing Nanoparticles, properties have to be studied. Despite all the above mentioned, the data on the potential environmental effects of nanoparticles are rare. The pipelines of pharmaceutical companies are believed to be drying up in many cases, and a number of blockbuster drugs will come off patent in the near-term. Using nanoparticles, it may be possible to achieve improved delivery of poorly water-soluble drugs by delivering drug in small particle size increase the total surface area of the drugs allowing faster dissolution in blood stream. Faster the dissolution translates in to faster absorption by human body targeted delivery of drugs in a cell- or tissue-specific manner. A nanoparticle has emerged as a promising strategy for the efficient delivery of drugs used for the treatment of some diseases by specific targeting. These carriers are designed in such a way that they are independent in the environments and selective at the pharmacological site. The formation of nanoparticle and physiochemical parameters such as pH, monomer concentration, ionic strength as well as surface charge, particle size and molecular weight are important for drug delivery. Further, these nanoparticles have the capability to reverse multidrug resistance a major problem in chemotherapy. Well-established therapies commonly employed in cancer treatment include surgery, Chemotherapy, immunotherapy, and radiotherapy. This mini-review is summarizing the information on different methods of manufacturing nanoparticles, focusing on its properties and biological transport.P acids are important active components in traditional medicines with different chemical structures and characters. Ferulic acid is a major phenolic acid found in numerous plant species. Because of the presence of phenolic group, it acts as a natural antioxidant, free radical scavenger and metal chelator, and could bind to enzymes and other multi subunit proteins, and modify their structure and function. Alpha-2-macroglobulin is a large glycoprotein present in the plasma as well as extravascular fluids of vertebrates. It is one of the major antiproteinases capable of inhibiting virtually any proteinase of any origin. Apart from being a major antiproteinase, α2M has many other diversified and complex functions, such as, binding, transportation and targeting. In the present study, effect of ferulic acid was observed on the structure of sheep α2M using various spectroscopy techniques, such as UV spectroscopy, fluorescence spectroscopy, circular dichroism (CD) and Fourier transform infrared spectroscopy (FTIR). With the help of UV and fluorescence spectroscopy, it was found that the conformation of antiproteinase has been changed due to the α2M-ferulic acid combination. CD and FTIR studies also revealed a change in secondary and tertiary structure of α2M in the presence of ferulic acid. From the results of these techniques, binding mode was discussed. Ahmed Abdur Rehman et al., Altern Integ Med 2013, 2:10 http://dx.doi.org/10.4172/2327-5162.S1.003Cisplatin (CP; cis-diamminedichloroplatinum II) is a major antineoplastic drug effective against a broad spectrum of malignancies. Patients receiving CP, however, experience very profound and long lasting gastrointestinal symptoms. Nigella sativa seeds/oil have been used in a wide range of gastrointestinal disorders. In view of this, the present study investigates the protective effect of Nigella sativa oil (NSO) on CP-induced adverse biochemical alterations in the small intestine of rat. Rats were divided into four groups, the CP+NSO and NSO groups were administered NSO (2ml/kg b.wt orally), with or without single dose of CP treatment (6mg/kg b. wt. i.p) respectively. Serum parameters, activities of brush border membrane (BBM) enzymes and various oxidative stress parameters were analysed in mucosal homogenates and/or brush border membrane vesicles (BBMV). CP treatment caused significant decline in the activities of alkaline phosphatase (ALP), γglutamyltransferase (GGTase), leucine aminopeptidase (LAP) and sucrase both in the mucosal homogenates and in the isolated brush border membrane vesicles (BBMV). However, the CP induced decrease in the activities of ALP, GGTase, LAP and sucrase was significantly prevented when CP treatment was extended to NSO administered rats. Lipid peroxidation and total sulfhydryl groups were altered upon CP treatment. CP significantly decreased the activities of SOD, catalase, GSH-Px, GR and TR. The CP induced changes in the activities of SOD, CAT and GSH-Px, GR and TR were ameliorated when CP treatment was given to NSO fed rats. The results suggest that NSO owing to its intrinsic biochemical/antioxidant properties enhanced resistance to CP elicited adverse gastrointestinal effects.M are part of a greater “sacred physiology”, that maps out the body according to subtle energy currents and power points which when manipulated can alter both the organic functions and structural conditions of the body. In Ayurveda, marmas which are used for the diagnosis and treatment of disease are basically pressure points meaning “vulnerable” or “sensitive zones” can also be used for promoting health and longevity. Various medical methods are identified of treating marmas like abhyanga (massage), aroma therapy, mardana, pranic healing, application of lepas, herbal treatments and various instrumental methods which can be helpful in treating pain, bleeding, swelling, restoring vital energy, etc. It can be used to balance the doshas, to increase agni (the digestive fire), for detoxification (reduce ama), as well as to promote vigour (vajikarana) and aid in rejuvenation (rasayana). These methods can be supplementary to other ayurvedic therapies, whether constitutional or disease focused, applied along with them for added enhancement.