Comparative assessment of organic and inorganic tea leaf extract feeding on anxiety behaviour status of colchicine-induced rat model of Alzheimer's disease.

Tea (Camellia sinensis), having anti-inflammatory, antioxidant, and free radical scavenging properties, may be beneficial to prevent the symptoms of neurodegenerative disorders like Alzheimer's disease (AD). In this present study, field experiments using the productive tea clone (TV25) with four nutrient management treatments were conducted during 2015 to 2017 in the research farm of Agricultural and Food Engineering Department, Indian Institute of Technology Kharagpur. The four nutrient management treatments were no application of fertilizer (control), organic fertilizer (OF), inorganic fertilizer (IF), and integration of OF and IF (IF + OF). The contents of different catechins of tea leaves grown under these treatments were measured using High Performance Liquid Chromatography. Tea leaf samples of these treatments were fed to the intracerebroventricular (ICV) colchicine administered rats. The animal study was double-blinded and randomized. Assessment of anxiety status was done for the rat model in an elevated open field with a novel object in two intervals (14-day and 21-day study). Anxiolytic behaviour with the lower corticosterone (CORT) level (82 ng/ml) was observed in ICV colchicine administered rat models of AD. After feeding of organically and inorganically grown tea extract (10, 20, and 30 mg/kg) for 14 days and 21 days, it was found that the anxiolytic behaviour decreased with the increased concentration of serum CORT. However, organic tea showed greater increase in CORT level (216.1 ng/ml) as compared to inorganic tea (214 ng/ml). Thus, this study showed organic tea may act as a favourable agent or adjuvant in the improvement of the anxiolytic behaviour in rat model of AD.

Dual Functionalized Injectable Hybrid Extracellular Matrix Hydrogel for Burn Wounds.

Low strength and rapid biodegradability of acellular dermal matrix (ADM) restrict its wider clinical application as a rapid cell delivery platform in situ for management of burn wounds. Herein, the extracted ADM was modified by a dual cross-linking approach with ionic crosslinking using chitosan and covalent cross-linking using an iodine-modified 2,5-dihydro-2,5-dimethoxy-furan cross-linker, termed as CsADM-Cl. In addition, inherent growth factors and cytokines were found to be preserved in CsADM-Cl, irrespective of ionic/covalent crosslinking. CsADM-Cl demonstrated improvement in post crosslinking stiffness with a decreased biodegradation rate. This hybrid crosslinked hydrogel supported adhesion, proliferation, and migration of human foreskin-derived fibroblasts and keratinocytes. Also, the angiogenic potential of CsADM-Cl was manifested by chick chorioallantoic membrane assay. CsADM-Cl showed excellent antibacterial activity against Escherichia coli and Staphylococcus aureus. Moreover, CsADM-Cl treated full thickness burn wounds and demonstrated rapid healing marked with superior angiogenesis, well-defined dermal-epidermal junctions, mature basket weave collagen deposition, and development of more pronounced secondary appendages. Altogether, the bioactive CsADM-Cl hydrogel established significant clinical potential to support wound healing as an apt injectable antibacterial matrix to encounter unmet challenges concerning critical burn wounds.

Comparative evaluation of therapeutic efficacy of intra-articular oxaceprol with conventional modalities in osteoarthritis animal model.

The duration and dose-dependent side effects of conventional intra-articular corticosteroid treatment in osteoarthritis (OA) like cartilage damage and chondrocyte toxicity warrant the search for alternative therapeutics. Oxaceprol, a recognized oral therapeutic agent for osteoarthritis, is yet to be explored for its intra-articular route of administration confirming better safety profile. In this study, a comparative evaluation of intra-articular oxaceprol and corticosteroid is carried out in osteoarthritis rabbit model. Osteoarthritis was induced by monosodium iodoacetate in rabbits. After randomization into three groups of five animals each: OA with intra-articular injection of saline, OA with intra-articular injection of oxaceprol, and OA with intra-articular injection of corticosteroids, treatment efficacy was analyzed by evaluation of inflammation through knee swelling, pain assessment by wire walking, and hot plate method. Further biopsies were collected for histological characterization. Intra-articular oxaceprol and corticosteroids reduced 20.5 and 24.5% knee swelling respectively within 4 weeks compared to those in control osteoarthritic rabbits. Oxaceprol exhibited analgesic action in visual analogue scoring of wire walking method. Hot plate test further confirmed drastic minimization of pain in oxaceprol intervention. Histological investigation suggested that application of oxaceprol has the abilities to protect articular cartilages from degenerative changes that occur in osteoarthritis. Marked improvement both in bone and cellular matrixes was observed in oxaceprol-treated group while gross lesions were visible and consisted of a well-demarcated area of cartilage erosion in control group. Intra-articular injection of oxaceprol showed remarkable improvement of articular cartilage in chemically induced osteoarthritic rabbits.

Novel Insights into the Molecular Interaction of a Panduratin A Derivative with the Non Structural Protein (NS3) of Dengue Serotypes: A Molecular Dynamics Study.

BACKGROUND: The ligand PKP10 having substitution of Cl- at R2 and R3 positions of ring A of Panduratin A i.e., ((1R,2S,5S)-5-(2,3-dichlorophenyl)-3-methyl-2-(3-methylbut-2-nyl)cyclohex-3- enyl)(2,6-dihydroxy-4-methylphenyl)methanone hydrate) has been observed to block the Nuclear Receptor Binding Protein binding site of Non Structural protein 3 in all dengue serotypes. In continuation with our earlier study, we have reported sixty novel Panduratin A derivatives compounds where substitution was done in positions 2 and 3 position of the benzyl ring A of Panduratin A with various substituents. METHODS: We selected ((1R,2S,5S)-5-(2,3-dichlorophenyl)-3-methyl-2-(3-methylbut-2-nyl)cyclohex-3- nyl) (2,6-dihydroxy-4-methylphenyl) methanone hydrate) (PKP10) for molecular dynamics (MD) simulations as it constantly produced lowest CDocker interaction energy of among all the sixty five derivatives. The CDocker interaction energy was predicted to be -140.804, -79.807, -78.217 and -84.073 Kcalmol-1 respectively against NS3 protein of dengue serotypes (DENV1-4). To understand the dynamics of the PKP10 with NS3 protein, each complex was subjected to molecular dynamics simulations of 50 ns in aqueous solution. MD (Molecular Dynamics) simulation study revealed that the binding of ligand PKP10 at the active site of NS3 induces a conformational change in all serotypes which was well supported by principal component analysis. RESULT: To the best of our knowledge, this is first ever study which provided atomistic insights into the interaction of PKP10 with NS3 protein of dengue serotypes. CONCLUSION: The result from our study along with in vitro studies is expected to open up better avenues to develop inhibitors for dengue virus in the near future.

Accelerated healing of full thickness dermal wounds by macroporous waterborne polyurethane-chitosan hydrogel scaffolds.

Wound healing is a dynamic process wherein cells, and macromolecules work in consonance to facilitate tissue regeneration and restore tissue integrity. In the case of full-thickness (FT) wounds, healing requires additional support from native or synthetic matrices to aid tissue regeneration. In particular, a matrix with optimum hydrophilic-hydrophobic balance which will undergo adequate swelling as well as reduce bacterial adhesion has remained elusive. In the present study, polyurethane diol dispersion (PUD) and the anti-bacterial chitosan (Chn) were blended in different ratios which self-organized to form macroporous hydrogel scaffolds (MHS) at room temperature on drying. SEM and AFM micrographs revealed the macroporosity on top and fracture surfaces of the MHS. FTIR spectra revealed the intermolecular as well as intra-molecular hydrogen bonding interactions between the two polymers responsible for phase separation, which was also observed by micrographs of blend solutions during the drying process. The effect of phase separation on mechanical properties and in vitro degradation (hydrolytic, enzymatic and pH dependent) of MHS were studied and found to be suitable for wound healing. In vitro cytocompatibility was demonstrated by the proliferation of primary rat fibroblast cells on MHS. Selected MHS was subjected to in vivo FT wound healing study in Wistar rats and compared with an analogous polyurethane containing commercial dressing i.e. Tegaderm™. The MHS-treated wounds demonstrated accelerated healing with increased wound contraction, higher collagen synthesis, and vascularization in wound area compared to Tegaderm™. Thus, it is concluded that the developed MHS is a promising candidate for application as FT wound healing dressings.

Assessment of Insulin Sensitivity and its Convalescence with Dietary Rehabilitation in Undernourished Rural West Bengal Population.

INTRODUCTION: Metabolic syndrome involving diabetes is acquiring the eminence of prospective epidemic in India with dramatic rise in rural population prevalence where majority are undernourished and creates significant healthcare burden. AIM: To investigate the prevalence of the insulin resistance amongst the undernourished rural population, involving children, adolescents and adult volunteers of the West Bengal and the effect of nutritional supplementation in the parameters echoing insulin sensitivity. MATERIALS AND METHODS: A prospective interventional study was carried out on 120 volunteers in three age groups; Group 1 (5-12 years), Group 2 (13-18 years) and Group 3 (19-40 years), each with 40 subjects who fulfilled the inclusion criteria, under informed consent over six months. Baseline data regarding demography, family history including parental history of diabetes, medical history, physical, activity, anthropometrical status and blood parameters echoing insulin sensitivity were obtained from volunteers. Intervention included daily nutritional supplementation with eggs and banana. After six months of food supplementation, parameters reflecting insulin sensitivity were assessed. RESULTS: Initial low figures of fasting insulin and blood sugar levels improvised with the nutritional supplementation alongside the absence of wasting in Group 1. HOMA-IR and IRI values of the Group 2 improved from the initial figures (3.19, 2.87) to (2.204, 1.99) and that of Group 3 from (5.0, 4.52) to (4.08, 3.67) respectively over six months (p<.05). Average Fasting Blood Sugar (FBS) enhanced from 85.5±9.6 mg/dl to 78.4±7.3 mg/dl and from 90.1±10.2 mg/dl to 84.8±8.4 mg/dl in Group 2 and 3 respectively by the end of the supplementation study (p<.05). CONCLUSION: Six months study revealed the existence of insulin resistance and malnourishment in the selected rural population in three groups. Decrement in HOMA-IR, IRI, FBS and serum insulin alongside the improvement in the body mass, in comparison to that of initial values is evocative of restoration of insulin sensitivity with nutrition supplementation.

Fenugreek, A Potent Hypoglycaemic Herb Can Cause Central Hypothyroidism Via Leptin - A Threat To Diabetes Phytotherapy.

Fenugreek (Trigonella foenum graecum), a medicinal herb with potent antihyperglycaemic and hypoglycaemic effects, is used to treat diabetes. This study is aimed to explore the interaction of fenugreek seed extract (FSE) and HPT (hypothalamic-pituitary-thyroid) axis in context of leptin secretion which have important role in normal and type-1 diabetic subjects. FSE (confirmed to contain trigonelline, diosgenin, 4 hydroxyisoleucine) was gavaged (0.25 gm/kg body weight/day) to normal and alloxan-induced type-1 diabetic rats for 4 weeks. Expression of hypothalamic prepro-TRH (Thyrotropin releasing hormone) mRNA, serum levels of TRH, TSH (Thyroid stimulating hormone), fT3, fT4, insulin, leptin, glucose; thyroperoxidase activity and growth of thyroid gland, food intake, adiposity index were also studied FSE significantly down regulated prepro-TRH mRNA expression; decreased serum TRH, TSH, fT3, fT4 levels, and regressed thyroid gland in FSE-fed normal and diabetic rats than those observed in normal diet-fed control and diabetic rats. FSE decreased (p<0.005-0.001) adiposity index and leptin secretion, increased food intake and body weight in all FSE-fed rats. FSE improved insulin secretion, decreased glucose level but impaired HPT axis in diabetic rats, indicating insulin-independent central hypothyroidism. Results suggested that the dominant signal to hypothalamus suppressing HPT axis is the fall in leptin level which i resulted from decreased adiposity index following FSE feeding. Fenugreek simultaneously having hypoglycaemic and hypothyroidal actions raises questions whether it can be safely used to treat diabetes and/or hyperthyroidism as was suggested by many workers.

In Vivo Hypoglycemic Studies of Polyherbal Phytoceuticals, Their Pharmacokinetic Studies and Dose Extrapolation by Allometric Scaling.

BACKGROUND: This work reports the safety profiling, in vivo hypoglycemic and pharmacokinetic studies of three phytoceuticals viz. conventional and sustained release tablets and microspheres each containing a polyherbal product phytocomposite (PHC) as the active ingredient. PHC is prepared from the leaf extracts of Ficus benghalensis: Syzigium cumini: Ocimum sanctum mixed in the weight ratio of 1:1:2. Further no observed adverse effect level (NOAEL), maximum recommended starting dose (MRSD) in human and prediction of human pharmacokinetic parameters have been accomplished by allometric equations. METHODS: Acute and sub chronic studies of the phytoceuticals were done as per OECD and in vivo hypoglycemic studies in STZ induced diabetic rats. Plasma concentrations of the active constituent rutin (pharmacologically active compound of PHC) were determined by HPLC and other pharmacokinetic parameters using PK Solver. Repeated dose toxicity was carried out to determine the NOAEL value, MRSD estimated using allometric formulas of body surface area and clearance (CL) and volume of distribution (Vd) predicted by allometric equations of single species scaling. RESULTS: Phytoceuticals showed a wide range of safety profile with a significant lowering of blood gluco-lipid level. The values of the pharmacokinetic parameters for different doses of phytoceuticals showed that the active concentration was maintained in plasma level and each formulation complied with their relevant quality criteria. NOAEL value was 5000 mg/kg/body weight and MRSD was 4864.86 mg. CONCLUSION: Phytoceuticals prepared are safe and effectively controlled blood gluco lipid level. Animal to human dose extrapolation and prediction of human pharmacokinetic parameters by allometry was convenient.

PBPK Modeling - A Predictive, Eco-Friendly, Bio-Waiver Tool for Drug Research.

BACKGROUND: The world has witnessed growing complexities in disease scenario influenced by the drastic changes in host-pathogen- environment triadic relation. Pharmaceutical R&Ds are in constant search of novel therapeutic entities to hasten transition of drug molecules from lab bench to patient bedside. Extensive animal studies and human pharmacokinetics are still the "gold standard" in investigational new drug research and bio-equivalency studies. Apart from cost, time and ethical issues on animal experimentation, burning questions arise relating to ecological disturbances, environmental hazards and biodiversity issues. Grave concerns arises when the adverse outcomes of continued studies on one particular disease on environment gives rise to several other pathogenic agents finally complicating the total scenario. Thus Pharma R&Ds face a challenge to develop bio-waiver protocols. Lead optimization, drug candidate selection with favorable pharmacokinetics and pharmacodynamics, toxicity assessment are vital steps in drug development. METHODS: Simulation tools like Gastro Plus™, PK Sim®, SimCyp find applications for the purpose. Advanced technologies like organ-on-a chip or human-on-a chip where a 3D representation of human organs and systems can mimic the related processes and activities, thereby linking them to major features of human biology can be successfully incorporated in the drug development tool box. RESULTS: PBPK provides the State of Art to serve as an optional of animal experimentation. PBPK models can successfully bypass bio-equivalency studies, predict bioavailability, drug interactions and on hyphenation with in vitro-in vivo correlation can be extrapolated to humans thus serving as bio-waiver. CONCLUSION: PBPK can serve as an eco-friendly bio-waiver predictive tool in drug development.

Rising trend of diabetes mellitus amongst the undernourished: State -of- the -art review.

Diabetes mellitus is prevailing in the malnourished populations congruently in well-nourished ones with an escalating trend in the former group regardless of the absence of obesity as an etiologic determining factor as per the studies in underprivileged sectors of the population. Chronic undernutrition across a lifetime may be an imperative stimulator of diabetes in an individual either by progressively reducing beta cell function alongside islet cell volume and increasing the individual predisposition to other genetic or environmental diabetogenic influences with modifying influence on the course of clinical syndrome. Ketosis resistant insulinopenia is irreversible to the sustained vigorous nutritional convalescence in a substantial fraction of malnourished subjects. It also debunks a latent diabetic stage with insulin resistance reflected by greater insulin requirement in comparison to the patients with type I diabetes with the same beta cell failure fraction and obese type II diabetic patients with equivalent glycemic control gauged by HbA1c levels. Current tendency warrants the replacement of conventional therapy by community oriented theranostic approaches and health programs to curb the epidemic.

Radiopaque Hemocompatible Ruminant-Sourced Gut Material with Antimicrobial Physiognomies for Biomedical Applications in Diabetics.

This study comprises the fabrication of a radiopaque gut material with its mechanical properties conforming to the US Pharmacopeia guidelines giving an antimicrobial advantage for suture application, especially in conditions such as diabetes mellitus, which has a high wound infection rate. Schiff base cross-linking iodination of the material is evinced by the spectroscopic studies, and antimicrobial properties owing to released iodine are evinced through in vitro studies. Modified gut sutures demonstrated favorable physicomechanical features such as appropriate tensile strength (440 ± 20 MPa) and knot strength (270 ± 20) alongside a mean radiopacity value of 139.0 ± 10 in comparison with that of the femoral shaft with 160 ± 10. The diabetic model showed absence of clinical signs of infection, supported by wound swab culture and the absence of necrosis in histology. Hemocompatibility studies evinced the absence of contact platelet activation and hemolysis alongside the customary coagulation response. These promising results highlight the stimulating potential of the process in the development of biomedical applications, necessitating persistent studies for its evidence-based applicability, particularly in diabetic patients.

Onion derived carbon nanodots for live cell imaging and accelerated skin wound healing.

Nitrogen, sulfur, and phosphorous co-doped water-soluble carbon nanodots are synthesized from culinary waste onion peel powder (OPP) by a short microwave treatment. Onion Derived Carbon Nano Dots (OCND) that comprised hydrophilic group-decorated amorphous nano-dots exhibited bright, stable fluorescence at an excitation of 450 nm and emission wavelength at 520 nm along with a free radical scavenging property. The OCND exhibited excellent stability at different pH and UV exposure. Although extracted polyphenols degraded in the extract, interestingly it was shown to be cytocompatible and blood compatible as observed during cytotoxicity, fluorescence imaging of the cell and a hemolysis study. The present work not only focuses on the synthesis of OCND from the OPP extract but also provides an interesting fact that, even after the degradation of polyphenols in the extract, they are non-toxic to human cells (HFF & MG63) and RBCs. Moreover, OCND had no adverse effect on the migration rate of Human Foreskin-derived Fibroblasts (HFFs) as observed from a scratch assay. In addition to accelerating the migration rate of fibroblasts, the OCND altered intra- and extracellular reactive oxygen species (ROS) by enhancing the antioxidant mechanism of a fibroblast under oxidative stress. Further, OCND was observed to accelerate wound healing in a full thickness (FT) wound in a rat model for topical application, which can be attributed to its radical scavenging potential. In summary, this study leads to a new type of OCND synthesis route, which is inherently co-doped with phosphorous, sulfur and nitrogen and holds a great promise for a myriad of biological applications, including bio-imaging, free radical scavenging and wound healing.

Computational pharmacokinetics and in vitro-in vivo correlation of anti-diabetic synergistic phyto-composite blend.

Despite tremendous strides in modern medicine stringent control over insulin resistance or restoration of normoglycemia has not yet been achieved. With the growth of molecular biology, omics technologies, docking studies, and in silico pharmacology, modulators of enzymes and receptors affecting the molecular pathogenesis of the disease are being considered as the latest targets for anti-diabetic therapy. Therapeutic molecular targets are now being developed basing on the up or down regulation of different signaling pathways affecting the disease. Phytosynergistic anti-diabetic therapy is in vogue both with classical and non-classical medicinal systems. However its chemo-profiling, structural and pharmacokinetic validation awaits providing recognition to such formulations for international acceptance. Translational health research with its focus on benchside product development and its sequential transition to patient bedside puts the pharma RDs to a challenge to develop bio-waiver protocols. Pharmacokinetic simulation models and establishment of in vitro-in vivo correlation can help to replace in vivo bioavailability studies and provide means of quality control for scale up and post approval modification. This review attempts to bring different shades highlighting phyto-synergy, molecular targeting of antidiabetic agents via different signaling pathways and bio-waiver studies under a single umbrella.

Top-Down and Bottom-Up Approaches in 3D Printing Technologies for Drug Delivery Challenges.

3-Dimensional printing (3DP) constitutes a raft of technologies, based on different physical mechanisms, that generate a 3-dimensional physical object from a digital model. Because of its rapid fabrication and precise geometry, 3DP has gained a prominent focus in biomedical and nanobiomaterials research. Despite advancements in targeted, controlled, and pulsatile drug delivery, the achievement of site-specific and disease-responsive drug release and stringent control over in vivo biodistribution, are still some of the important, challenging areas for pharmaceutical research and development and existing drug delivery techniques. Microelectronic industries are capable of generating nano-/microdrug delivery devices at high throughputs with a highly precise control over design. Successful miniaturizations of micro-pumps with multireservoir architectures for delivery of pharmaceuticals developed by micro-electromechanical systems technology were more acceptable than implantable devices. Inkjet printing technologies, which dispense a precise amount of polymer ink solutions, find applications in controlled drug delivery. Bioelectronic products have revolutionized drug delivery technologies. Designing nanoparticles by nanoimprint lithography showed a controlled drug release pattern, biodistribution, and in vivo transport. This review highlights the "top-down" and "bottom-up" approaches of the most promising 3DP technologies and their broader applications in biomedical and therapeutic drug delivery, with critical assessment of its merits, demerits, and intellectual property rights challenges.

In vitro-in vivo studies of the quantitative effect of calcium, multivitamins and milk on single dose ciprofloxacin bioavailability.

Ciprofloxacin, commonly used in India as an anti-microbial for prolonged use in chronic and non-specific indications, may affect the bioavailability of the drug. The drug prescribed is commonly taken with multivitamins, calcium and milk. A simple and reliable analytical methodology obtaining a correlation with in vivo urinary excretion studies using UV and HPLC and in vitro dissolution studies (IVIVC) has shown a significant increase in elimination rate of ciprofloxacin co-administered with multivitamins, calcium and milk. Appreciable IVIVC results proved that dissolution studies could serve as an alternative to in vivo bioavailability and also support bio-waivers.

Chemometrics Optimized Extraction Procedures, Phytosynergistic Blending and in vitro Screening of Natural Enzyme Inhibitors Amongst Leaves of Tulsi, Banyan and Jamun.

BACKGROUND: Tulsi, Banyan, and Jamun are popular Indian medicinal plants with notable hypoglycemic potentials. Now the work reports chemo-profiling of the three species with in-vitro screening approach for natural enzyme inhibitors (NEIs) against enzymes pathogenic for type 2 diabetes. Further along with the chemometrics optimized extraction process technology, phyto-synergistic studies of the composite polyherbal blends have also been reported. OBJECTIVE: Chemometrically optimized extraction procedures, ratios of polyherbal composites to achieve phyto-synergistic actions, and in-vitro screening of NEIs amongst leaves of Tulsi, Banyan, and Jamun. MATERIALS AND METHODS: The extraction process parameters of the leaves of three plant species (Ficus benghalensis, Syzigium cumini and Ocimum sanctum) were optimized by rotatable central composite design of chemometrics so as to get maximal yield of bio-actives. Phyto-blends of three species were prepared so as to achieve synergistic antidiabetic and antioxidant potentials and the ratios were optimized by chemometrics. Next, for in vitro screening of natural enzyme inhibitors the individual leaf extracts as well as composite blends were subjected to assay procedures to see their inhibitory potentials against the enzymes pathogenic in type 2 diabetes. The antioxidant potentials were also estimated by DPPH radical scavenging, ABTS, FRAP and Dot Blot assay. RESULTS: Considering response surface methodology studies and from the solutions obtained using desirability function, it was found that hydro-ethanolic or methanolic solvent ratio of 52.46 ± 1.6 and at a temperature of 20.17 ± 0.6 gave an optimum yield of polyphenols with minimal chlorophyll leaching. The species also showed the presence of glycosides, alkaloids, and saponins. Composites in the ratios of 1:1:1 and 1:1:2 gave synergistic effects in terms of polyphenol yield and anti-oxidant potentials. All composites (1:1:1, 1:2:1, 2:1:1, 1:1:2) showed synergistic anti-oxidant actions. Inhibitory activities against the targeted enzymes expressed in terms of IC50 values have shown that hydro-ethanolic extracts in all cases whether individual species or composites in varying ratios gave higher IC50 values thus showing greater effectivity. CONCLUSION: Current research provides the state-of-the-art of search of NEIs amongst three species by in-vitro assays which can be further utilized for bioactivity-guided isolations of such enzyme inhibitors. Further, it reports the optimized phyto-blend ratios so as to achieve synergistic anti-oxidative actions. SUMMARY: The current research work focuses on the optimization of the extraction process parameters and the ratios of phyto-synergistic blends of the leaves of three common medicinal plants viz. banyan, jamun and tulsi by chemometrics. Qualitative and quantitative chemo profiling of the extracts were done by different phytochemical tests and UV spectrophotometric methods. Enzymes like alpha amylase, alpha glucosidase, aldose reductase, dipeptidyl peptidase 4, angiotensin converting enzymes are found to be pathogenic in type 2 diabetes. In vitro screening of natural enzyme inhibitors amongst individual extracts and composite blends were carried out by different assay procedures and the potency expressed in terms of IC50 values. Antioxidant potentials were estimated by DPPH radical scavenging, ABTS, FRAP and Dot Blot assay. Hydroalcoholic solvent (50:50) gave maximal yield of bio-actives with minimal chlorophyll leaching. Hydroethanolic extract of tulsi showed maximal antioxidant effect. Though all composites showed synergism, maximal effects were shown by the composite (1:1:2) in terms of polyphenol yield, antioxidant effect and inhibitory actions against the targeted enzymes. Abbreviations used: DPP4- dipeptidyl peptidase 4; AR- aldose reductase; ACE- angiotensin converting enzyme; PPAR-γ- peroxisome proliferator activated receptor-γ; NEIs- natural enzyme inhibitors; BE- binding energy; GLP-1- Glucagon like peptide -1; ROS- Reactive oxygen species; CAT- catalase; GSH-Px- glutathione per-oxidase; SOD- superoxide dismutase; pNPG- para-nitro phenyl-α-D-gluco-pyranoside solution; DPPH- 1,1-diphenyl-2-picrylhydrazyl; RSM- Response surface methodology; CCD- central composite design; DMSO- dimethyl sulfoxide; HHL- hippuryl-L-histidyl-L-leucine; GPN-Tos- Gly-Pro p-nitroanilide toluenesulfonate salt; ESC- experimental scavenging capacity; TSC- theoretical scavenging capacity; FRAP- Ferric Reducing Assay Procedure; ABTS- 2, 2'- azinobis (3-ethylbenzothiazoline-6 - sulfonic acid.

In vitro-in vivo studies of the quantitative effect of calcium, multivitamins and milk on single dose ciprofloxacin bioavailability / 药物分析学报

Ciprofloxacin, commonly used in India as an anti-microbial for prolonged use in chronic and non-specific indications, may affect the bioavailability of the drug. The drug prescribed is commonly taken with multivitamins, calcium and milk. A simple and reliable analytical methodology obtaining a correlation with in vivo urinary excretion studies using UV and HPLC and in vitro dissolution studies (IVIVC) has shown a significant increase in elimination rate of ciprofloxacin co-administered with multivitamins, calcium and milk. Appreciable IVIVC results proved that dissolution studies could serve as an alternative to in vivo bioavailability and also support bio-waivers.

Automated tissue classification framework for reproducible chronic wound assessment.

The aim of this paper was to develop a computer assisted tissue classification (granulation, necrotic, and slough) scheme for chronic wound (CW) evaluation using medical image processing and statistical machine learning techniques. The red-green-blue (RGB) wound images grabbed by normal digital camera were first transformed into HSI (hue, saturation, and intensity) color space and subsequently the "S" component of HSI color channels was selected as it provided higher contrast. Wound areas from 6 different types of CW were segmented from whole images using fuzzy divergence based thresholding by minimizing edge ambiguity. A set of color and textural features describing granulation, necrotic, and slough tissues in the segmented wound area were extracted using various mathematical techniques. Finally, statistical learning algorithms, namely, Bayesian classification and support vector machine (SVM), were trained and tested for wound tissue classification in different CW images. The performance of the wound area segmentation protocol was further validated by ground truth images labeled by clinical experts. It was observed that SVM with 3rd order polynomial kernel provided the highest accuracies, that is, 86.94%, 90.47%, and 75.53%, for classifying granulation, slough, and necrotic tissues, respectively. The proposed automated tissue classification technique achieved the highest overall accuracy, that is, 87.61%, with highest kappa statistic value (0.793).

Exploration of natural enzyme inhibitors with hypoglycemic potentials amongst Eucalyptus Spp. by in vitro assays.

AIM: To investigate the presence and potency of natural enzyme inhibitors with hypoglycemic potentials amongst Eucalyptus Spp. by in vitro assays. METHODS: The leaf extracts of the three different Eucalyptus species [E. globulus (EG), E. citriodora (EC), E. camaldulensis (ECA)] were subjected to in vitro assay procedures to explore the prevalence of natural enzyme inhibitors (NEIs) after preliminary qualitative and quantitative phytochemical evaluations, to study their inhibitory actions against the enzymes like α-amylase, α-glucosidase, aldose reductase, angiotensin converting enzyme and dipeptidyl peptidase 4 playing pathogenic roles in type 2 diabetes. The antioxidant potential and total antioxidant capacity of the species were also evaluated. RESULTS: Major bioactive compounds like polyphenols (341.75 ± 3.63 to 496.85 ± 3.98) and flavonoids (4.89 ± 0.01 to 7.15 ± 0.02) were found in appreciable quantity in three species. Based on the IC50 values of the extracts under investigation, in all assays the effectivity was in the order of EG > ECA > EC. The results of the ferric reducing antioxidant power assay showed that the reducing ability of the species was also in the order of EG > ECA > EC. A strong correlation (R(2) = 0.81-0.99) was found between the phenolic contents and the inhibitory potentials of the extracts against the targeted enzymes. CONCLUSION: These results show immense hypoglycemic potentiality of the Eucalyptus Spp. and a remarkable source of NEIs for a future phytotherapeutic approach in Type 2 diabetes.

Frequency dependent impedimetric cytotoxic evaluation of anticancer drug on breast cancer cell.

The present work reports the impedance characteristics of MCF-7 cell lines treated with anticancer drug ZD6474 to evaluate the cytotoxic effect on cellular electrical behaviour using miniature impedance sensors. Four types of impedance sensing devices with different electrode geometries are fabricated by microfabrication technology. The frequency response characteristics of drug treated cells are studied to evaluate cytotoxic effect of ZD6474 and also to assess the frequency dependent sensitivity variation with electrode area. A significant variation in magnitude of measured impedance data is obtained for drug treated samples above 10 µM dose indicating prominent effect of ZD6474 which results in suppression of cell proliferation and induction of apoptosis process. The results obtained by impedimetric method are correlated well with conventional in vitro assays such as flow cytometry, cell viability assays and microscopic imaging. Finally an empirical relation between cell impedance, electrode area and drug dose is established from impedance data which exhibits a negative correlation between drug doses and impedance of cancer cells.