Invitro Method Research Articles

Collagen IV of basement membranes: III. Chloride pressure is a primordial innovation that drives and maintains the assembly of scaffolds

Collagen IV scaffold is a primordial innovation enabling the assembly of a fundamental architectural unit of epithelial tissues-a basement membrane attached to polarized cells. A family of six α-chains (α1 to α6) coassemble into three distinct protomers that form supramolecular scaffolds, noted as collagen IVα121, collagen IVα345, and collagen IVα121-α556. Chloride ions play a pivotal role in scaffold assembly, based on studies of NC1 hexamers from mammalian tissues. First, Cl- activates a molecular switch within trimeric NC1 domains that initiates protomer oligomerization, forming an NC1 hexamer between adjoining protomers. Second, Cl- stabilizes the hexamer structure. Whether this Cl--dependent mechanism is of fundamental importance in animal evolution is unknown. Here, we developed a simple invitro method of SDS-PAGE to determine the role of solution Cl- in hexamer stability. Hexamers were characterized from 34 animal species across 15 major phyla, including the basal Cnidarian and Ctenophora phyla. We found that solution Cl- stabilized the quaternary hexamer structure across all phyla except Ctenophora, Ecdysozoa, and Rotifera. Further analysis of hexamers from peroxidasin knockout mice, a model for decreasing hexamer crosslinks, showed that solution Cl- also stabilized the hexamer surface conformation. The presence of sufficient chloride concentration in solution or "chloride pressure" dynamically maintains the native form of the hexamer. Collectively, our findings revealed that chloride pressure on the outside of cells is a primordial innovation that drives and maintains the quaternary and conformational structure of NC1 hexamers of collagen IV scaffolds.

Comparing and evaluating the obturation techniques by 3 type of delivery system: An invitro method

: The purpose of this study was to compare in vitro three methods of root canal obturation technique in deciduous dentition using conventional radiographic technique.: 45 root canals of primary molars were prepared by regular manual technique and obturated using zinc oxide eugenol and calcium hydroxide-based syringe form. Obturation methods compared were lentulo spiral, plugger and Iodontin syringe form. Conventional radiographic technique was used to evaluate the Root canal length, Obturation and the presence of voids. The obtained data were analyzed using Chi-square test.The results showed significant differences between the three groups for the radioopacity of obturation (P < 0.05). The results showed Iodontin syringe form has fewer voids compared to lentulo spiral ZOE paste and Plugger for obturation quality. Statistically significant difference was found between Iodontin syringe form and Plugger technique (p= <0.05).: Iodontin syringe form showed the best results in the length of obturation and as well as controlling paste extrusion through the apical foramen. The lentulo spiral and plugger can also be used as an alternative method.

Systematic Review on Herbs with Antidandruff Property

Abstract: The most common problem in teenage and adult is of dandruff which is scientifically known as ‘Malassezia’ which if ignore can further continue with secondary symptoms like Acne, itching, discomfort etc. The present review highlighted the mechanism of ‘Malassezia’ along with the available treatment for it. Though the number of synthetic formulations available, the present review focuses on alterative and safe way to treat dandruff i.e. use of herbs with their biological source and constituents responsible for effectivity to solve the problem in broad population. Due to wide range of activity phytoconstituents like anti-inflammatory, antibacterial and antifungal, these synergistic effect may increase the applicability of herbs in cosmetics and cosmeceuticals as well. Hence the considering the need of herbal formulation development an attempt has been made to discuss about formulation commonly used to treat dandruff i.e. shampoo, different parameters of evaluation and invitro method evaluation of Antidandruff activity.

Development of the Polyphenol Compounds from Plant Raw Materials by the Cell Micropropagation IN VITRO Method

One of the ways to design polyfunctional ingredients that determine the functional properties of food products are complex food additives consisting of valuable plant raw materials. The additives quality depends on the used raw materials adequacy, time and collection place, and the plant cultivation conditions. The traditional plantation method requires significant costs to obtain secondary metabolites determining physiological plant materials value. The research aim is to obtain polyphenolic compounds from phyto-raw materials by microclonal cell propagation under sterile laboratory conditions. The leaves and berries cells of cowberries and cranberries containing a significant number of polyphenols are the research objects. For example, cranberries growing in the south of the Tyumen region contain anthocyanins 97.8 mg/100 g and leukoanthocyanins 459.6 mg/100 g, and berries growing in the Arctic territories of the Yamalo-Nenets Autonomous Okrug contain 224.7 and 480.2 mg/100 g, respectively. At the initial stage, the researchers sterilized objects, instruments, and equipment. They determined the sterilization duration experimentally. The study revealed that with a longer sterilization duration, there was a change in color and an increase in cell toxicity, with a minimum duration, pathogenic microorganisms remained in the medium. After sterilization, a man cultured the cells in the light under fluorescent lamps with illumination of 100 mmol quanta/m²; photoperiod of 20 h per day; in nutrient media with an acidity of pH 5.2–5.4; 25 ml in volume; enriched with the auxins and cytokinins hormones. The researchers used Kundu phytohormones to regulate the secondary metabolites synthesis. They utilized Murashige Skoog and Anderson agar media to find the optimal nutrient medium in order to obtain the maximum increase in polyphenolic compounds in plant cells. A man developed the third agar media independently, considering the prospects of an individual approach to the more efficient producers growth.

Evaluation of the Role of Merromoside from Ipomoea aquatica Forsskal Hydroalcoholic Extract in the Downregulation of ROS Species in Overcoming MDR in Breast Cancer.

The medicinal plant Ipomoea aquatica belonging to convulvulaceae family is an effective natural herb for treatment of various ailments and possesses effective anticancer activity. The aim of the work is to characterize a secondary metabolite merromoside (a resin glycoside) for anti-breast cancer activity through down regulation of ROS species. The Extract of the whole plant has been prepared by maceration method using 50%v/v ethanol in distilled water to get a hydroalcoholic extract. The phytochemical evaluation reveals that the active secondary metabolite was isolated by using column chromatographic technique. The isolated compound was evaluated for its anticancer properties through invitro method such as 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay on Michigan Cancer Foundation-7 Cell lines. The purity and structural characterization were done by high-performance thin layer chromatography, Fourier Transform Infrared Spectroscopy, Proton and13C Nuclear Magnetic Resonance spectroscopy and Liquid Chromatography-Mass Spectrometry. The isolated compound (W04) from the derived extract showed Rf value of 0.79 that showed IC50 of 182.8μg/ml. The chemical structure of W04 has been confirmed as [4,5-dihydroxy-6-[5-hydroxy-2-methyl-4-(2-methylpropanoyloxy)-6-[(24,25,26-trihydroxy-5,23-dimethyl-9-oxo-19-pentyl-2,4,8,20,22-pentaoxatricyclo[19.2.2.13,7]hexacosan-6-yl)oxy]oxan-3-yl]oxy-2-methyloxan-3-yl] 2-methyl propanoate with the molecular weight of 979.15268. The isolated compound merromoside from hydroalcoholic extract of Ipomoea aquatica has been evaluated for anti-breast cancer properties. The down regulation of ROS species will prevent reverse signalling and angiogenesis. This indicates that merromoside will overcome MDR in breast cancer especially DOX-resistant.

Comparative gene expression analysis for pre-osteoblast MC3T3-E1 cells under non-adhesive culture toward osteocyte differentiation

Osteocytes play an important role to modulate the bone remodeling and are also known as terminally differentiated cells originated from the osteoblast precursor cells, but its differentiation mechanism remains unclear. Since an efficient invitro method to evoke the osteocyte differentiation from the osteoblast precursor cells has not been established, we conducted the comparative gene expression analysis for mouse pre-osteoblast MC3T3-E1 cells in order to elucidate the key factors to induce the osteocyte differentiation from the pre-osteoblast cells. In this study, we prepared four different culture environments by modulating their cell-substrate interaction and cell-cell interaction; (i) low and (ii) high cell density on the adhesive culture models, and (iii) low and (iv) high cell density on the non-adhesive floating culture models. By comparing these conditions in terms of cell-substrate and cell-cell interaction, we showed that the elimination of cell-substrate interaction under non-adhesive floating culture greatly up-regulated the gene expression of osteocyte markers in the pre-osteoblast cells. Moreover, the presence of moderate cell-cell interaction in the non-adhesive spheroid culture further enhanced the up-regulation of osteocyte markers for the pre-osteoblast cells. The results altogether suggest the most appropriate culture environment to induce the invitro osteocyte differentiation of pre-osteoblast cells.

Antioxidant and Skeletal Muscle Relaxant Activity of Leaf Extract of Plant Piper Attenuatum (B. HAM)

Piper attenuatum (B. Ham) is traditional medicinal plant in India. It has been claimed in traditional Indian system of medicine that the phytochemical constituents present in P. attenuatum (B. Ham) have Antioxidant and skeletal muscle relaxant activity. So the present study aimed to evaluate anti–oxidant and skeletal muscle relaxant activity of ethanolic, aqueous and ethanolic leaf extract of plant P. attenuatum (B. Ham), respectively. Antioxidant activity was evaluated by in vitro as well as in vivo methods. Invitro method we used DPPH (2, 2-diphenyl-1-picrylhydrazyl) free radical scavenging assay and H2O2 (Hydrogen Peroxide) scavenging assay while in vivo methods antioxidant enzymes like Superoxide Dismutase (SOD) and reduced glutathione (GSH) were assayed by using animal models. Anti-oxidant activity of following concentrations 10μg/mL, 20μg/mL, 30μg/mL, 40μg/mL and 50μg/mL were measured for both extracts. Ethanolic extract of P. attenuatum (B. Ham) was evaluated at dose of 100mg/kg & 200 mg/kg b.w. for skeletal muscle relaxant activity by using rota rod apparatus. The high antioxidant activity was found in the ethanolic extract of P. attenuatum (B. Ham) compared to aqueous one. For muscle relaxation, 200 mg/kg b.w. showed a significant reduction in the time spent by the animals on the revolving rod compared to the control. From the above study it may be concluded that both extract of P. attenuatum (B. Ham) having Anti – oxidant and skeletal muscle relaxant activity.

Sugar composition and in vitro glycaemic response of selected sugars, rice-based meals and traditional kuih

High glycaemic index foods were found to increase the risk of getting diabetes, obesity, cancer and cardiovascular disease. This study aimed to determine the in vitro glycaemic response and sugar composition of selected food types, which include sugars, rice-based meals and traditional cakes and desserts (kuih). The food samples were purchased prepared and analysed using High Performance Liquid Chromatography (HPLC) to determine the sugar composition. The samples were also prepared and assayed using validated in vitro digestion method to determine the estimated glycaemic response. The in vitro method used 0.25g total available carbohydrate, which was established by Clegg Anthrone method. Among the studied samples, the total sugar content was icing sugar (100%), which has the highest percentage, nasi kerabu (7.49%) and kuih lapis (42.20%) for sugars, rice-based meals and traditional kuih, respectively. Dialyzable glucose concentration for the sugar samples was highest in the sugar in sachet (3.47 mg/mL) and lowest in the sugar with stevia (0.49 mg/mL). For the different types of rice-based meals, the highest dialyzable glucose concentration was in nasi lemak (13.06 mg/mL), and the lowest was in nasi kerabu (12.33 mg/mL). While for traditional kuih, kuih lapis (10.53 mg/mL) and kuih keria (6.30 mg/mL) were highest and lowest in the dialyzable glucose concentration, respectively. Significant correlations exist between dialyzable glucose concentration (at 120 minutes) and sugar composition (p<0.05) for the rice-based samples. Among the traditional kuih samples, sucrose content and total sugar content was correlated significantly. However, no correlation exists between dialyzable glucose concentration and sugar composition for the selected sugar samples. Factors such as particle size and presence of phytochemical in sugars, available nutrients (starch, protein and fat) in kuih and cooked dishes as well as the amount of added sugar in food are vital factors that affect the in vitro glycaemic response of the studied samples.

Validation of a predictive method for sunscreen formula evaluation using gene expression analysis in a Chinese reconstructed full‐thickness skin model

This study aimed to establish a predictive invitro method for assessing the photoprotective properties of sunscreens using a reconstructed full-thickness skin model. A full-thickness skin model reconstructed with human fibroblasts and keratinocytes isolated from Chinese skin was exposed to daily UV radiation (DUVR). We examined the transcriptomic response, identifying genes for which expression was modulated by DUVR in a dose-dependent manner. We then validated the methodology for efficacy evaluation of different sunscreens formulas. The reconstructed skin model was histologically consistent with human skin, and upon DUVR exposure, the constituent fibroblasts and keratinocytes exhibited transcriptomic alterations in pathways associated with oxidative stress, inflammation and extracellular matrix remodelling. When used to evaluate sunscreen protection on the model, the observed level of protection from UV-induced gene expression was consistent with the corresponding protection factors determined clinically and allowed for statistical ranking of sunscreen efficacy. Within this study we show that quantification of gene modulation within the reconstructed skin model is a biologically relevant approach with sensitivity and predictability to evaluate photoprotection products.

In Vitro – In Vivo Evaluation of Antiurolithiatic activity of piperine from Piper nigrum

Background: A natural product is a organic compound or substance produced by a living organism that is, found in nature. Natural products show significant pharmacological or biological activity that can be of therapeutic benefit in treating diseases. Now a day’s Urolithiasis or Urinary calculi or Kidney stone formation becoming very prevalent in the world and it is reported that Urolithiasis is the 3rd most prevalent disease among the kidney diseases. Therefore there is an immediate urge in searching for alternative treatment for urolithiasis. Fruits of Piper longum Linn were commonly called as Black pepper have a long history in Indian traditional medicine and in Ayurveda for the treatment of Gastrointestinal and Respiratory complications. 1-(5(1,2-benzodioxol-5-yl)-1-oxo-2,4-pentadieneyl) piperidine commonly known as piperine were reported to possess many pharmacological activities. Purpose: This study evaluated the effect of Piperine on anti-urolithiatic activity in invitro and invivo models. Method: The anti-urolithiatic activity of Piperine was evaluated by using invitro methods like titrimetric method and aggregation assay. In vivo studies were done using male wistar rats. Results: The results of this study proved that Piperine has a significant anti-urolithiatic activity in rats (in-vivo) as well as in in-vitro models. Two test doses of piperine (40,80 mg/kg P.O) are evaluated using urolithiasis induced rats in in-vivo, titrimetric and aggregation in in-vitro models and it showed significant inhibition of crystallization with a significance of p<0.01 and p<0.05 when compared with the standard drug cystone (750mg/kg P.O.) Different serum parameters such as calcium, urea, uric acid, creatinine and urine parameters such as calcium and oxalate are assessed to evaluate anti urolithiatic activity of piperine in in-vivo study. The results were presented as mean±SEM. Difference among data was statistically analysed using One-way ANOVA to determine the level of significance using Graph pad Prism Differences between the data were considered significant at P < 0.05 and P<0.01. Conclusion: In in-vitro method of evaluation, piperine (20mg/kg) demonstrated a significant anti-urolithiatic activity than piperine (10mg/kg) when compared with a standard drug Cystone and in in-vivo models, piperine (40mg/kg) demonstrated a significant anti- urolithiatic activity than piperine (80mg/kg) when compared with a standard drug Cystone.

Photoprotection against visible light-induced pigmentation.

This paper presents invivo an invitro studies demonstrating the induction of pigmentation in human skin by visible light which can be blocked by using formulation containing the correct amount of yellow iron oxide (YIO). An invitro absorption method was developed to determine the protection provided by a test formulation containing 4.5% YIO using an IPD UVA-VIS action spectrum. Following the development of the invitro method and invivo study with 10 normal healthy volunteers with Fitzpatrick skin phototypes IV to VI was conducted to verify if the predictive model. The invitro model for visible light protection provided a protection factor of 2.5 using the invitro absorption spectrum of 4.5% of YIO with a very similar result from the invivo study with a protection factor of 3.0. Multiple daily exposures of visible light have shown increase in skin pigmentation and the application of YIO provide less development of pigmentation when compared to unprotected skin. In vitro testing of the absorbance of the pigmented formulation using a proposed action spectrum for immediate pigment darkening (IPD) response in the visible light range supports the invivo protection observations for persistent pigment darkening (PPD) and can be used as predictor for skin pigmentation induced by visible light.

Discovering Synergistic Drug Combination from a Computational Perspective.

Synergistic drug combinations play an important role in the treatment of complex diseases. The identification of effective drug combination is vital to further reduce the side effects and improve therapeutic efficiency. In previous years, in vitro method has been the main route to discover synergistic drug combinations. However, many limitations of time and resource consumption lie within the in vitro method. Therefore, with the rapid development of computational models and the explosive growth of large and phenotypic data, computational methods for discovering synergistic drug combinations are an efficient and promising tool and contribute to precision medicine. It is the key of computational methods how to construct the computational model. Different computational strategies generate different performance. In this review, the recent advancements in computational methods for predicting effective drug combination are concluded from multiple aspects. First, various datasets utilized to discover synergistic drug combinations are summarized. Second, we discussed feature-based approaches and partitioned these methods into two classes including feature-based methods in terms of similarity measure, and feature-based methods in terms of machine learning. Third, we discussed network-based approaches for uncovering synergistic drug combinations. Finally, we analyzed and prospected computational methods for predicting effective drug combinations.

Development and Validation of a Test to Identify Drugs That Cause Idiosyncratic Drug-Induced Liver Injury

Idiosyncratic drug-induced liver injury (iDILI) is one of the most challenging diagnoses in hepatology. It is frequently impossible to identify the agent that has caused iDILI in patients who take multiple medicines. We developed an invitro method to identify drugs that cause liver injury in patients, based on drug toxicity to monocyte-derived hepatocyte-like (MH) cells from patient blood samples. We then collected data on patients who were re-exposed to drugs found to be toxic in the MH test to validate test performance. We performed a prospective study of patients referred to the University Hospital in Munich,Germany, with acute liver injury believed to be caused by medications (300 patients were enrolled in the study and we present data from 40 patients with iDILI and re-exposure to implicated drugs). We collected data from patients on medical history, laboratory testand imaging results, findings from biopsy analyses, and medications taken. Blood samples were collected from all patients and MH cells were isolated and cultured for 10days. MH cells were then incubated with drugs to which each patient had been exposed, and toxicity was measured based on release of lactate dehydrogenase. Agents found to be toxic to MH cells were considered as candidates for the cause of liver injury. Patients were followed up for up to 6 months after liver injury and data on drug re-exposures and subsequent liver damage within the following 3 to 24 months were associated with findings from MH tests. Our test identified 10 drugs that were toxic to MH cells from 13 patients (amoxicillin/clavulanateto cells from 2 patients; diclofenac to cells from 2 patients; methylprednisolone to cells from 2 patients; and atorvastatin, metamizole, pembrolizumab, piperacillin/tazobactam, moxifloxacin, duloxetine, or sertraline each to cells from 1 patient). Thirteen patients had a recurrence of liver injury after inadvertent re-exposure to a single drug, and the MH test correctly identified 12 of the 13 drugs that caused these liver re-injury events. All 86 drugs that were not toxic to MH cells in our assay were safely resumed by patients and were not associated with liver re-injury in 27 patients. Therefore, the MH test identifies drugs that cause liver injury with 92.3% sensitivity and 100% specificity (1 false-negative and 12 true-positive results). We developed a test to identify drugs that cause liver injury in patients based on their toxicity to MH cells isolated from patients with DILI. We validated results from the assay and found it to identify drugs that cause DILI with 92.3% sensitivity and 100% specificity. The MH cell test could be a tool to identify causes of iDILI, even in patients taking multiple medications. ClinicalTrials.gov no: NCT02353455.

Age-Associated Impairments in Mitochondrial ADP Sensitivity Contribute to Redox Stress in Senescent Human Skeletal Muscle

It remains unknown if mitochondrial bioenergetics are altered with aging in humans. We established an invitro method to simultaneously determine mitochondrial respiration and H2O2 emission in skeletal muscle tissue across a range of biologically relevant ADP concentrations. Using this approach, we provide evidence that, although the capacity for mitochondrial H2O2 emission is not increased with aging, mitochondrial ADP sensitivity is impaired. This resulted in an increase in mitochondrial H2O2 and the fraction of electron leak to H2O2, in the presence of virtually all ADP concentrations examined. Moreover, although prolonged resistance training in older individuals increased muscle mass, strength, and maximal mitochondrial respiration, exercise training did not alter H2O2 emission rates in the presence of ADP, the fraction of electron leak to H2O2, or the redox state of the muscle. These data establish that a reduction in mitochondrial ADP sensitivity increases mitochondrial H2O2 emission and contributes to age-associated redox stress.

DISSOLUTION METHOD DEVELOPMENT WITH CHROMATOGRAPHIC METHOD FOR DETERMINATION OF DRUG RELEASE IN DISSOLUTION SAMPLES OF URSODEOXYCHOLIC ACID TABLETS.

Ursodeoxycholic Acid is used in the treatment of Liver disease. An accurate and robust invitro method is developed and validated for measurement of drug release in Ursodeoxycholic Acid tablets. HPLC method for quantification of drug in dissolution samples of Ursodeoxycholic Acid tablet is developed and validated. Phosphate buffer (0.05M) of pH- 7.5 is used as dissolution medium and paddle (USP-II) as apparatus at 75 rpm. The sample was withdrawn after 60 minutes. The developed HPLC method was used for quantitative estimation of drug release in dissolution samples of Ursodeoxycholic Acid tablet. The HPLC column of Hypersil - C18, 250X4.6 mm, 5µm is used. The phosphate buffer of pH – 6.5 and acetonitrile was mixed in the ratio of 50:50v/v pumped at a flow rate of 1.0 ml/minute. The detection was made at wavelength of 210 nm. Through method validation, it is proved that the method is accurate, rugged, precise and robust. The method is linear over the concentration range of 50% – 150%. The method also found very precise. Mean recovery of Ursodeoxycholic Acid was found 99.6 ±38%. Keywords: Ursodeoxycholic Acid, Dissolution, chromatography, Method Validation.

Effects of heat treatment on protein feeds evaluated invitro by the method of estimating utilisable crude protein at the duodenum.

The effect of heat treatment on the protein value of field beans, lupins and peas was studied using an invitro method. Protein feeds were subjected to heat treatment for 30, 60 and 90min in forced air oven at 120, 140 and 160°C and in autoclave at 105, 120 and 135°C. The heat-treated protein feeds were incubated in buffered rumen fluid together with grass silage and barley in complete isonitrogenous diets. The gas production (GP) was recorded continuously, and ammonia-N (NH3 -N) concentrations were determined during the invitro incubation at 8, 24 and 48h and used to determine the utilisable crude protein (uCP) at the duodenum at 16h of incubation (uCP16 ). Heat treatments decreased the concentration of soluble crude protein and increased neutral detergent insoluble CP (NDICP) in all protein feeds compared to untreated. Inclusion of protein feeds to basal diet showed no increase in the uCP16 in untreated field bean diet and only a small numerical increase in the uCP16 concentrations from 160g/kg dry matter (DM) to 166 and 172g/kg DM in untreated lupine and pea diets, respectively, indicating high degradability of untreated feeds. Increasing the time and temperature of the heat treatment linearly increased the uCP16 concentrations in field bean and pea diets, but not in lupin diets. Autoclave treatment was more effective in decreasing uCP16 than oven treatment despite the lower temperatures used. However, the combination of highest temperatures and treatment time in autoclave increased acid detergent insoluble CP (ADICP) concentrations in protein feeds, indicating protein damage and decreased intestinal digestibility. Determining invitro uCP and ADICP shows to be a promising method for evaluating protein value in heat-treated animal feeds.

PRELIMINARY PHYTOCHEMICAL SCREENING AND IN-VITRO FREE RADICAL SCAVENGING ACTIVITY OF ROOT EXTRACTS OF GLYCYRRHIZA GLABRA L

ABSTRACT: Obesity is an international health problem caused by various environmental and genetic factors. Herbal supplements have proved to be active against obesity. Glycyrrhiza glabra belongs to the family of Fabaceae. Glycyrrhiza glabra was used traditionally to treat various diseases. Hence the current study was planned to investigate the phytochemicals and antioxidants present in Glycyrrhiza glabra. Materials and Methods: The preliminary qualitative, quantitative phytochemicals and antioxidants were investigated from the root extracts of Glycyrrhiza glabra. Results: The preliminary qualitative analysis of various extracts of Glycyrrhiza glabra showed the presence of Phenols, Flavonoids, Tannins, Saponins and Quinones. Quantitative analysis of Glycyrrhiza glabra showed elevated amounts of phenols, flavonoids and tannins in ethanolic extracts. The Aqueous, ethanol, hydro alcoholic extracts of Glycyrrhiza glabra showed 211±2.64, 281.66±1.57, 263±14.2 mg of phenols, 152.66±3.05, 187.33±1.52, 183.33±3.05 mg of flavonoids, 21.33±0.57, 41±3.46, 34.3±1.15 mg of Tannins, 42.33±2.05, 63.33±1.52, 36.33±1.57 mg of terpenoids. The antioxidant properties of the ethanolic extract of Glycyrrhiza glabra were evaluated with the 2,2-diphenyl-1-picryl hydrazyl (DPPH), Nitric Oxide radical (NO), Superoxide radical (SO), Hydrogen peroxide radical (H2O2), Hydroxyl radical (HO) Scavenging activity. The IC50 value of Glycyrrhiza glabra is 120 µg/ml for DPPH, 208 µg/ml for (NO), 196 µg/ml for (SO), 148 µg/ml for (H2O2), and 252 µg/ml for (OH). Invitro method suggests that Glycyrrhiza glabra is more powerful against nitric oxide and hydroxyl radicals. Conclusion: This preliminary study shows that Glycyrrhiza glabra is a promising herb with many phytochemicals and antioxidants it can be further investigated for its antiobesity potential.Key words: Glycyrrhiza glabra L., phytochemicals, antioxidant and obesity.

Evaluation of Hemidesmus indicus root as an antacid by In vitro

Medicinal plants are being widely used, either as single drug or in combination in health care delivery system. Indian Sarsaparilla, Hemidesmus indicus (Family: Asclepiadaceae) is a commonly known Indian Medicinal Plant, which is widely recognized in traditional systems of Medicine. It contains various phytoconstituents belonging to the category glycosides, flavonoids, tannins, sterols and volatile oils. It has been reported as useful in biliousness, blood diseases, dysentery, diarrhea, respiratory disorders, skin diseases, syphilis, fever, leprosy, leucoderma, leucorrhoea, itching, bronchitis, asthma, eye diseases, epileptic fits in children, kidney and urinary disorders, loss of appetite, burning sensation, dyspepsia, nutritional disorders, ulcer and rheumatism. Several studies are being carried towards its activities like analgesic, anti-inflammatory, antiulcer, hepatoprotective, antioxidant and helicobactericidal properties. In our study we have evaluated antacid activity of sariva (Anantmool) by using In-Vitro method, i.e. ANC (Acid Neutralizing Capacity). This evaluation was done by comparing the ANC of sariva macerated &amp; powdered drug with water as blank &amp; standard drug i.e. NaHCO3. Based on this In-Vitro experiment, we can conclude that, the macerated &amp; powdered drug of sariva (Anantmool) evaluated in this study, varied in potency as measured in terms of their ANC. These results having ** i.e. P &lt; 0.01 &amp; Passed the normality test. However, the present study being in-vitro, the effects of antacid may vary In-Vitro; individual variations also contribute to the ultimate effectiveness of as antacid.

Verification on the use of the Inoue method for precisely determining glomerular filtration rate in Philippine pediatrics

The objective of this study is to validate the Inoue method so that it would be the preferential choice in determining glomerular filtration rate (GFR) in Philippine pediatrics. The study consisted of 36 patients ranging from ages 2 months to 19 years old. The subjects used were those who were previously subjected to in-vitro method. The scintigrams of the invitro method was obtained and processed for split percentage uptake and for parameters needed to obtain Inoue GFR. The result of this paper correlates the Inoue GFR and In-vitro method (r = 0.926). Thus, Inoue method is a viable, simple, and practical technique in determining GFR in pediatric patients.

Generation of Functional Thymic Epithelium from Human Embryonic Stem Cells that Supports Host T Cell Development

Inducing immune tolerance to prevent rejection is a key step toward successful engraftment of stem-cell-derived tissue in a clinical setting. Using human pluripotent stem cells to generate thymic epithelial cells (TECs) capable of supporting Tcell development represents a promising approach to reach this goal; however, progress toward generating functional TECs has been limited. Here, we describe a robust invitro method to direct differentiation of human embryonic stem cells (hESCs) into thymic epithelial progenitors (TEPs) by precise regulation of TGFβ, BMP4, RA, Wnt, Shh, and FGF signaling. The hESC-derived TEPs further mature into functional TECs that support Tcell development upon transplantation into thymus-deficient mice. Importantly, the engrafted TEPs produce Tcells capable of invitro proliferation as well as invivo immune responses. Thus, hESC-derived TEP grafts may have broad applications for enhancing engraftment in cell-based therapies as well as restoring age- and stress-related thymic decline.