• Title/Summary/Keyword: formulation assay

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In-Vitro Anticancer and Free Radical Scavenging Potential of Compound Formulation Used in Unani System of Medicine

  • Mannan, Mohd Nazir;Kazmi, Munawwar Husain;Chakraborty, Alokananda;Zakir, Mohammad;Ahmad, Tasleem;Lahari, K.
    • CELLMED
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    • v.10 no.4
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    • pp.27.1-27.6
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    • 2020
  • Cancer is one of the leading cause of mortality in India as well as worldwide. The management of cancer by conventional therapy has shown life threatening adverse effects. The researchers are now exploring the natural way of treatment. Unani system of medicine have rich literature for cancer and many compound formulations have been described in this system. Unani system of medicine is based on holistic approach and treat human being as a unit with natural herbs, mineral and animal origin drugs. An important compound Unani formulation (CUF) from the literature has been chosen to explore the Unani claim of its anticancer activity. The phytochemical constituents were assessed using standard phytochemical screening method. Antioxidant property of this formulation was assessed by DPPH assay. The DPPH free radical scavenging assay was carried out by colorimetric method and ascorbic acid was taken as a positive control. Three different extracts of CUF on different concentrations were used to screening on human breast cancer (BCC) MCF-7 cell line. For the estimation of in-vitro cytotoxic potency of the investigated extracts was assessed on MTT assay by using trypan blue method and paclitaxel was used as the standard. Hydro-ethanolic (HE) extract showed highest free radical scavenging activity among all extracts. DPPH Assay showed substantial antioxidant activity of these extracts in hydro-ethanol extract at 1㎍ concentration of CUF. The CUF showed antioxidant and anticancer activity. The claim made by Unani physician has been proved.

Effects of a Powder Formulation of Streptomyces cameroonensis on Growth and Resistance of Two Cocoa Hybrids from Cameroon against Phytophthora megakarya (Causal Agent of Black Pod Disease)

  • Aristide, Dzelamonyuy;Martial, Tene Tayo Paul;Ruth, Ngotcho Ngassam Esther;Grace, Lele Brenda;Ebenezer, Foka Tatiekam;Flore, Magni Pacha Tatiana;Thaddee, Boudjeko
    • Journal of Microbiology and Biotechnology
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    • v.32 no.2
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    • pp.160-169
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    • 2022
  • In the present study we evaluated the efficacy of a bioformulation of Streptomyces cameroonensis for control of black pod disease in cocoa and enhancement of seedling growth. The formulation developed using talc powder and cassava starch as carriers showed high shelf-life of 1.07 × 106 CFU/g after six months storage at 4℃. The formulation was tested for inhibition of spore germination in Phytophthora megakarya and showed 100% inhibition at 10% (w/v) of formulation. To determine the efficacy of the formulation, we performed an in planta assay in the greenhouse on two hybrids of cocoa seedlings, the tolerant SNK413 × (♂) T79/467 and the susceptible UPA 134× (♂) SCA 12. Detached leaf assay showed a significant reduction in the disease severity index of about 67% for the tolerant hybrid and 55% for the susceptible hybrid compared to non-treated plants. A significant enhancement in stem length, leaf surface area and root weight was observed. Analysis of biochemical markers of defense showed a significant increase in total polyphenol, flavonoid, and total protein contents. There was also significant upregulation of PR-proteins such as chitinases, peroxidases and β-1, 3-glucanases following treatment of both tolerant and susceptible hybrids, though with a higher level of synthesis in the tolerant hybrids. A significant increase was also observed in polyphenol oxidase activities in plants treated with the formulation. This work demonstrated the stability and effectiveness of the S. cameroonensis powder formulation in suppressing black pod disease in cocoa and subsequently enhancing the growth of seedlings.

Antidiabetic Activity of an Ayurvedic Formulation Chaturmukha Rasa: A Mechanism Based Study

  • Sharma, Akansha;Tiwari, Raj K;Sharma, Vikas;Pandey, Ravindra K;Shukla, Shiv Shnakar
    • Journal of Pharmacopuncture
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    • v.22 no.2
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    • pp.115-121
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    • 2019
  • Objectives: The objective of this study was to evaluate antidiabetic activity of Chaturmukha rasa based on streptozotocin induced diabetes model, alpha amylase inhibitory activity, alpha Glucosidase inhibitory activity and inhibition of sucrase. Methods: Chaturmukha rasa was prepared as per Ayurvedic formulary. Antidiabetic activity was measured in experimentally streptozotocin induced rats. The dose was taken as 45 mg/kg, i.p. The antidiabetic activity of Chaturmukha rasa was compared Triphala Kwatha, a marketed formulation. Further In vitro $\acute{\alpha}$- Amylase Inhibitory Assay, In vitro salivary amylase Inhibitory Assay, In vitro ${\alpha}-Glucosidase$ Inhibitory Assay and In vitro Sucrase Inhibitory Assay was performed with respect to Chaturmukha rasa. The IC50 value was calculated for all the above activity. Results: Streptozotocin with Acarbose showed significant decrease in blood glucose level whereas streptozotocin with Triphala kwatha showed more decrease in blood glucose level than Streptozotocin with Acarbose. The combination of Streptozotocin + Triphala kwatha + Chaturmukha rasa showed a significant decrease in blood glucose level on 21st day. In vitro $\acute{\alpha}$- Amylase Inhibitory Assay the Chaturmukha rasa showed IC50 value $495.94{\mu}l$ when compared with Acarbose $427.33{\mu}l$, respectively. In the ${\alpha}-Glucosidase$ Inhibitory Assay Chaturmukha rasa showed IC50 value $70.93{\mu}l$ when compared with Acarbose $102.28{\mu}l$, respectively. In vitro Sucrase Inhibitory Assay Chaturmukha rasa showed IC50 value $415.4{\mu}l$ when compared with Acarbose $371.43{\mu}l$, respectively. Conclusion: This study supports that Chaturmukha rasa may inhibit diabetes by inhibition of salivary amylase or alpha Glucosidase or sucrase. This may be the mechanism by which Chaturmukha rasa inhibits diabetes. Further this study supports the usage of Chaturmukha rasa for the management of diabetes.

Anti-bacterial effect of fusion formulation of Coptis rhizoma and Pelargonium sidoides on the growth of bronchial diseases bacteria (황련과 Pelargonium sidoides 복합제제의 호흡기 감염 세균에 대한 항균 효과)

  • Lee, Jong Rok;Min, Byung-Gu;Park, Chung A;Kim, Sang Chan;Park, Sook Jahr
    • Herbal Formula Science
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    • v.25 no.4
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    • pp.449-456
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    • 2017
  • Objective : Coptis rhizoma is traditional herb in Korean medicine, and Pelargonium sidoides extract has been used for relief of acute bronchitis in Western medicine. The present study examined the antibacterial effect of fusion formulation of Coptis rhizoma extract and Pelargonium sidoides extract against bronchial diseases bacteria. Methods : Test sample, fusion formulation of Korean and Western medicine, was prepared by mixing Coptis rhizoma extract and Pelargonium sidoides extract at a ratio of 1:2 (w/w). Antimicrobial properties of test sample were determined by agar diffusion assay and minimum inhibitory concentration (MIC) against bronchus diseases bacteria. Results : In agar diffusion assay, the highest amount of test sample (4 mg/disk) exhibited antibacterial activity against all microorganisms tested. Test sample showed the high activity for S. aureus (19.5 mm), C. diptheriae (16.5 mm), A. fumigatus (19.3 mm), F. nucleatum (22.7 mm) and Mycobactrium sp. (17.3 mm), whereas it showed a low activity for K. pneumonia (9.7 mm). The MIC value was determined as $250{\mu}g/m{\ell}$ against C. diptheriae. Test sample showed better growth inhibitory effects against S. aureus and A. fumigatus with the MIC valus of $125{\mu}g/m{\ell}$. Conclusion : These results suggest the possibility of application to chronic respiratory diseases of fusion formulation of Korean and Western medicine, which was prepared with Coptis rhizoma extract and Pelargonium sidoides extract.

Antioxidative Activity and Protection of Oxidative Chromosomal Damage by Vegetables, Fruits Extract and Their Functional Liquid Formulation (야채 및 과일추출물의 항산화작용과 산화적 염색체손상에 대한 억제효과)

  • 이승철;허찬;이승현;김현표;허문영
    • YAKHAK HOEJI
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    • v.48 no.2
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    • pp.111-116
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    • 2004
  • The ethanol extracts of mixed vegetables (Bioactive Vegetables, BV), mixed fruits (Bioactive Fruits, BF) and their liquid formulation (Chungpae Plus , CP) were evaluated for their antioxidative and antigenotoxic activities. They were shown to possess the significant free radical scavenging effect against 1,1-diphenyl-2-picryl hydrazine (DPPH) radical generation and were revealed to show the inhibitory effect of lipid peroxidation as measured by malondialdehyde (MDA) formation although the potencies were not higher than those of vitamin C. They did not possess any pro-oxidant effect on bleomycin-Fe(III) dependent DNA degradation, whereas vitamin C showed strong pro-oxidant effect. Furthermore, oral administration of BV and BF inhibited micronucleated reticulocyte (MNRET) formation of mouse peripheral blood induced by KBrO3 treatment in vivo. CP also showed significant inhibition under same experimental condition. Therefore, the liquid formulation (CP) containing BV and BF may be a useful natural antioxidative and antigenotoxic agent by scavenging free radicals, inhibition of lipid peroxidation and protecting chromosomal damage.

Investigation of Pharmacological Stability of Hyeonggaeyeongyotang Gagambang by Formulation Changes for Foaming Tablet (형개연교탕(荊芥連翹湯) 가감방(가감방)의 발포정 제형 변화에 따른 약리학적 안정성 연구)

  • Cho, Nam Joon;Gang, Seong Gu;Kim, Ji Young;Han, Hyo Sang;Kim, Kee Kwang
    • The Korea Journal of Herbology
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    • v.33 no.5
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    • pp.67-72
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    • 2018
  • Objectives : Natural extracts have been extensively studied to replace single agent drugs that cause a variety of side effects. However, studies of changes to the formulation of natural extracts has not been nearly proceed. We aimed to investigate whether pharmacological stability of hyeonggaeyeongyotang gagambang (HYT) is altered by formulation changes for foaming tablet. Methods : In this study, we performed freeze - drying of HYT, which is known to have antioxidant and anti - inflammatory properties, and then changed the formulation by foaming. Results : As a result, the foaming reaction appeared normally when HYT foamed tablets were put into water, and almost all of the substances were dissolved in the aqueous solution. In addition, we confirmed using high-performance liquid chromatograph that the geniposide used as an indicator material of HYT was stable in most of the formulations. It was confirmed that the change of HYT formulation did not affect the antioxidant efficacy by the 2,2'-azino-bis-3-ethylbenzothiazoline-6-sulphonic acid assay. Moreover, quantitative real-time PCR confirmed that the inhibitory effect of HYT on IL-$1{\beta}$ mRNA expression induced by lipopolysaccharides treatment in murine macrophage RAW 264.7 cells was similar in the solution of foaming tablet. Conclusions : These results suggest that the materials with various pharmacological effects can be stably maintained even when the formulation is changed by the foaming action of HYT. Our results are expected to provide important basic knowledge on formulation changes using various natural extracts.

Development of Modified Phenylalanine Ammonia-lyase for the Treatment of Phenylketonuria

  • Kim, Woo-Mi
    • Biomolecules & Therapeutics
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    • v.17 no.1
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    • pp.104-110
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    • 2009
  • Phenylketonuria (PKU) is an inherited metabolic disorder caused by mutations in the phenylalanine catabolic enzyme, phenylalanine hydroxylase (PAH). The use of phenylalanine ammonia-lase (PAL) by oral and parenteral routes as a therapeutic drug for PKU has been severely limited due to inactivation by intestinal proteolysis and immune reactions. PEGylation was applied to PAL to reduce the degrees of antigenicity and proteolytic inactivation. Kinetic experiments with native PAL and pegylated PALs were performed, and pH stability, temperature stability, and protease susceptibility were evaluated. Enzyme linked immunosorbent assay (ELISA) was carried out to measure the immune complex between pegylated PALs and antiserum that had been extracted from a PAL-immunized mouse. Pegylated PAL, especially branched pegylated PAL (10 kDa, 1:32), was more active for phenylalanine and more stable in pancreatic proteases than native PAL. Native PAL was optimal at pH 8.5, corresponding to the average pH range of the small intestine; the same finding was noted for pegylated PALs. All linear and branched pegylated PALs had low reactivity with mouse antiserum, especially the 1:16 formulation with linear 5-kDa PEG and the 1:32 formulation with branched 10-kDa PEG. Therefore, we suggest the 1:32 formulation with branched 10-kDa PEG as the most promising formulation for enzyme replacement therapy.

Cosmetic Efficacy of Supercritical Cannabis sativa Seed Extracts and Enhancement of Skin Permeation (초임계 대마종자 추출물의 화장품 효능과 경피흡수증진 효과)

  • Lee, Kwang Won;Park, Shinsung;Park, Su In;Shin, Moon Sam
    • The Journal of the Convergence on Culture Technology
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    • v.7 no.4
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    • pp.683-691
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    • 2021
  • The purpose of this study is to measure the yield and to evaluate the physiological activity of Cannabis sativa seed(Hemp seed) extracts extracted using a density fluctuation supercritical carbon dioxide for each temperature condition-30℃(HSSE30), 45℃(HSSE45), 60℃(HSSE60), and to enable dissolution of the poorly water-soluble extracts by liposome formulation and to enhance the skin permeability. As a result of the yield measurement, HSSE60 showed the highest yield, and in the antioxidant activities, HSSE45 had the highest total polyphenol content, and showed the highest DPPH, ABTS+ radical scavenging activities at the highest concentration of the extracts. As a result of the antimicrobial susceptibility testing, a clear zone appeared only in the Propionibateium acnes strain. It was confirmed that particle size was reduced and the absolute value of the zeta potential increased in the case of the formulation in which the extracts were in liposomes than in the formulation in which the extracts were dissolved in deionized water, and the skin permeability was improved. Based on these experimental results, we confirmed the possibility of using the hemp seed supercritical carbon dioxide extracts, a poorly water-soluble extract, can be applied as a functional natural material for cosmetics.

Determination of Carbon Source Utilization of Bacillus and Pythium Species by Biolog$^{(R)}$ Microplate Assay

  • Chun, Se-Chul;R.W. Schneider;Chung, Ill-Min
    • Journal of Microbiology
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    • v.41 no.3
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    • pp.252-258
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    • 2003
  • The carbon utilizations of Bacillus species and Pythium species were investigated by using a Biolog$^{(R)}$ microplate assay to determine if there are differences in the carbon utilizations of selected strains of these species. It may be possible to afford a competitive advantage to bacterial biological control agents by providing them with a substrate that they can readily use as a carbon source, for example, in a seed coating formulation. Microplates, identified as SFP, SFN and YT were used to identify spore-forming bacteria, nonspore-forming bacteria, and yeast, respectively. Bacterial and mycelial suspensions were adjusted to turbidities of 0.10 to 0.11 at 600 nm. One hundred microliters of each of the bacterial and mycelial suspension were inoculated into each well of each of the three types of microplates. L-arabinose, D-galactose, D-melezitose and D-melibiose of the 147 carbohydrates tested were found to be utilized only by bacteria, and not by Pythium species, by Biolog$^{(R)}$ microplate assay, and this was confirmed by traditional shake flask culture. Thus, it indicated that the Biolog$^{(R)}$ microplate assay could be readily used to search for specific carbon sources that could be utilized to increase the abilities of bacterial biological control agents to adapt to contrived environments.

Cosmetic Effect of Mixed Plant Extracts Including Saururus Chinensis, Morus Bombycis Stem and Morus Papyrifera Stem (삼백초, 뽕나무 줄기, 닥나무 줄기 등을 함유한 복합수목추출물의 화장품소재 특성)

  • Jeong, Hyang Li;Kim, Hyun Woo;Kim, Ji Hye;Kim, Jin Hong;Kim, Donguk
    • Korean Chemical Engineering Research
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    • v.50 no.4
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    • pp.610-613
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    • 2012
  • In this study, plant extracts including 25% Saururus chinensis, 20% Morus bombycis stem and 20% Morus papyrifera stem were tested to use as possible cosmetic agent. As cosmetic effect test, cell toxicity test, elastase inhibition assay, tyrosinase inhibition assay, anti-oxidation assay and temperature stability test were done as cosmetic assays. Plant extract showed low cell toxicity, excellent elastase inhibition and anti-oxidation effect, however, tyrosinase inhibition effect was lower. Skin, lotion and essence formulation was made using plant extracts and temperature stability test was done. Skin and essence indicated good stability, however, lotion showed phase separation and severe viscosity variation, which means lotion formulation change. From the research, plant extracts including Saururus chinensis, Morus bombycis stem and Morus papyrifera stem were suitable for possible cosmetic agent.