Search results for: Bioassay

Authors: Jeff Clarine, Chang-Shyh Peng, Daisy Sang

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Bioassay is the measurement of the potency of a chemical substance by its effect on a living animal or plant tissue. Bioassay data and chemical structures from pharmacokinetic and drug metabolism screening are mined from and housed in multiple databases. Bioassay prediction is calculated accordingly to determine further advancement. This paper proposes a four-step preprocessing of datasets for improving the bioassay predictions. The first step is instance selection in which dataset is categorized into training, testing, and validation sets. The second step is discretization that partitions the data in consideration of accuracy vs. precision. The third step is normalization where data are normalized between 0 and 1 for subsequent machine learning processing. The fourth step is feature selection where key chemical properties and attributes are generated. The streamlined results are then analyzed for the prediction of effectiveness by various machine learning algorithms including Pipeline Pilot, R, Weka, and Excel. Experiments and evaluations reveal the effectiveness of various combination of preprocessing steps and machine learning algorithms in more consistent and accurate prediction.

Keywords: Bioassay, machine learning, preprocessing, virtual screen.

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Authors: Tatiana L. Gorbunova

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The research is dedicated to the study of the polluted river inflow impact on the Black Sea coastal marine environment within the watercourse’s plumes in the Greater Sochi area applying bioassay methods with using freshwater and marine microalgae. River waters were analyzed using microalgae Chlorella vulgaris Beijer and sea waters were tested with marine diatoms Phaeodactylum tricornutum Bohlin. Experiments included algae cells abundancy growth assessments in acute (24 hours), sub-acute (72 hours) and chronic (168 hours/7 days) tests. The increase in algal cell growth rates compared to the control in the summer period was detected as a consequence of the recreational activities intensification during the tourism seasonal peak. Most of the analyzed samples demonstrated a significant effect of algae cells growth stimulation compared to the control. It is established that under the impact of contaminants carried by river’s drain to the sea, the capacity of the coastal marine ecosystem is partially capable to compensate its effect on the coastal biocenosis, but the general trends of the impact processes remain constant.

Keywords: Algae abundance growth, bioassay, microalgae, modeling.

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Authors: Etty Riani, Endang Gumbira-Said, Kashwar Syamsu, Kustiariyah, Kaseno, Muhammad Reza Cordova

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Sandfish is one of marine biota that has a biomedicine (bioactive compound) potency. People in Gorontalo Province, Indonesia, have been sandfish as an aphrodisiac for men as it is believed that sandfish has a steroid hormone potency. This research aims at studying using the steroid hormone potency from every fraction of sandfish (meat and innards) and its activity of male reproduction (rooster) as an aphrodisiac. Steroid extraction was done using Touchstone and Kasparow method, and then it was utilized to study the effectiveness of bioassay of rooster. This research had five treatments and was done in complete randomized design. Based on Lieberman-Burchard and bioassay test, the author found that sandfish extract contains steroid hormone. Sandfish extract was able to enrich testosterone and cholesterol concentration in blood serum; fastening secondary reproduction characteristics of the rooster, and increasing growth as well as improving rooster’s comb. Therefore, sandfish steroid is potential to be used as an aphrodisiac for men.

Keywords: Aphrodisiac, sandfish, secondary reproduction characteristic, steroid.

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Authors: Kittiwann Nimkerdphol, Masahiro Nakagawa

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Biological reactions of individuals of a testing animal to toxic substance are unique and can be used as an indication of the existing of toxic substance. However, to distinguish such phenomenon need a very complicate system and even more complicate to analyze data in 3 dimensional. In this paper, a system to evaluate in vitro biological activities to acute toxicity of stochastic self-affine non-stationary signal of 3D goldfish swimming by using fractal analysis is introduced. Regular digital camcorders are utilized by proposed algorithm 3DCCPC to effectively capture and construct 3D movements of the fish. A Critical Exponent Method (CEM) has been adopted as a fractal estimator. The hypothesis was that the swimming of goldfish to acute toxic would show the fractal property which related to the toxic concentration. The experimental results supported the hypothesis by showing that the swimming of goldfish under the different toxic concentration has fractal properties. It also shows that the fractal dimension of the swimming related to the pH value of FD Ôëê 0.26pH + 0.05. With the proposed system, the fish is allowed to swim freely in all direction to react to the toxic. In addition, the trajectories are precisely evaluated by fractal analysis with critical exponent method and hence the results exhibit with much higher degree of confidence.

Keywords: 3D locomotion, bioassay, critical exponent method, CEM, fractal analysis, goldfish.

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Authors: Rizna Triana Dewi, Sanro Tachibana, Ahmad Darmawan

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The bioassay-guided isolation and purification of an ethyl acetate extract of Aspergillus terreus MC751 led to the characterization of butyrolactone I as an antidiabetic and antioxidant. The antidiabetic activity of butyrolactone I was evaluated by α- glucosidase and α-amylase inhibition assays. Butyrolactone I demonstrated significant concentration-dependent, mixed-type inhibitory activity against yeast α-glucosidase with an IC50 of 54μM. However, the compound exhibited less activity against rat intestinal α-glucosidase and α-amylase. This is the first report on α-glucosidase inhibitory activity of butyrolactone I. The antioxidative activity of butyrolactone I was evaluated based on scavenging effects on 1,1- diphenyl-2-picrylhydrazyl (DPPH) (IC50 =51 μM) and hydrogen peroxide (IC50= 141 μM) radicals as well as a reducing power assay. The results suggest that butyrolactone I is a promising antidiabetic as well as antioxidant and should be considered for clinical trials.

Keywords: Aspergillus terreus MC751, antidiabetic, antioxidant, Butyrolactone I.

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Authors: N. Messaadia, D. Harzallah

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The effects of divers carbon substrates were investigated for the tabtoxin production of an isolated pathogenic Pseudomonas syringae pv. tabaci, the causal agent of wildfire of tobacco and are discussed in relation to the bacterium growth. The isolated organism was grown in batch culture on Woolley's medium (28°C, 200 rpm, during 5 days). The growth has been measured by the optical density (OD) at 620 nm and the tabtoxin production quantified by Escherichia coli (K-12) bioassay technique. The growth and the tabtoxin production were both influenced by the substrates (sugars, amino acids, organic acids) used, each, as a sole carbon source and as a supplement for the same amino acids. The most significant quantities of tabtoxin were obtained in presence of some amino acids used as sole carbon source and/or as supplement.

Keywords: Amino acid supplement, carbon substrates, batch culture, Pseudomonas syringae pv. tabaci.

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Authors: N. Singh, N. Sharma, J. K. Katnoria

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Water a vital component for all living forms is derived from variety of sources, including surface water (rivers, lakes, reservoirs and ponds) and ground water (aquifers). Over the years of time, water bodies are subjected to human interference regularly resulting in deterioration of water quality. Therefore, pollution of water bodies has become matter of global concern. As the water quality closely relate to human health, water analysis before usage is of immense importance. Improper management of water bodies can cause serious problems in availability and quality of water. The quality of water may be described according to their physico-chemical and microbiological characteristics. For effective maintenance of water quality through appropriate control measures, continuous monitoring of metals, physico-chemical and biological parameter is essential for the establishment of baseline data for the water quality in any study area. The present study has focused on to explore the status of water pollution in various areas and to estimate the magnitude of its toxicity using different bioassay.

Keywords: Genotoxicity, Heavy metals, Mutagenicity, Physico-chemical analysis.

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Authors: Hanan Moawad, Naglaa M. Abd EL-Rahman

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Erwinia carotovora var. carotovora is the main cause of soft rot in potatoes. Hyphaene thebaica was studied for biocontrol of E. carotovora which inhibited growth of E. carotovora on solid medium, a comparative study of classical and ultrasound-assisted extractions of Hyphaene thebaica fruit. The use of ultrasound decreased significant the total time of treatment and increase the total amount of crude extract. The crude extract was subjected to determine the in vitro, by a bioassay technique revealed that the treatment of paper disks with ultrasound extraction of Hyphaene thebaica reduced the growth of pathogen and produced inhibition zones up to 38mm in diameter. The antioxidant activity of ultrasound-ethanolic extract of Doum fruits (Hyphaene thebaica) was determined. Data obtained showed that the extract contains the secondary metabolites such as Tannins, Saponin, Flavonoids, Phenols, Steroids, Terpenoids, Glycosides and Alkaloids.

Keywords: Ultrasound, classical extract, Biological control, Erwinia carotovora, Hyphaene thebaica.

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Authors: M. M. Mon, S. S. Maw, Z. K. Oo

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Antioxidant activities of ethanolic extracts of Ardisia japonica Blume., Ageartum conyzoides Linn., and Cocculus hirsutus Linn Diels. leaves was determined qualitatively and quantitatively in this research. 1, 1-diphenyl-2-picrylhydrazyl (DPPH) free radical solution was used to investigate free radical scavenging activity of these leaves extracts. Ascorbic acid (Vitamin C) was used as the standard. In the present investigation, it is found that all of these extracts have remarkable antioxidant activities. The EC50 values of these ethanolic extracts were 12.72 μg/ml for A. japonica, 15.19 μg/ml for A. conyzoides, 10.68 μg/ml for C. hirsutus respectively. Among these Myanmar medicinal plants, C. hirsutus showed higher antioxidant activities as well as free radical scavenging activity than black tea (Camellia sinensis), the famous antioxidant, and A. japonica and A. conyzoides showed a rather lower antioxidant activity than tea extracts. According to results from bioassay with carrot discs infected with Agrobacterium tumefaciens, all extracts showed anti-tumor activity after 3 weeks of incubation. No gall was detected in carrot disks treated with C. hirsutus and A. japonica extracts in the dose of 100ppm and in carrot discs treated with A. conyzoides extract in the dose of 1000 ppm. Therefore, the research clearly indicates that these weedy plants of dry farm land are exceptionally advantageous for human health.

Keywords: Antioxidant, Anti-tumor activity, Carrot-discbioassay, DPPH

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Authors: Harshani Uggallage, Kapila D. Dissanayaka

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A Sulforhodamine-B assay-guided fractionation on Nigella sativa seeds was conducted to determine the presence of cytotoxic compounds against human hepatoma (HepG2) cells. Initially, a freeze-dried sample of Nigella sativa seeds was sequentially extracted into solvents of increasing polarities. Crude extracts from the sequential extraction of Nigella sativa seeds in chloroform and ethyl acetate showed the highest cytotoxicity. The combined mixture of these two extracts was subjected to bioassay guided fractionation using a modified Kupchan method of partitioning, followed by Sephadex® LH-20 chromatography. This chromatographic separation process resulted in a column fraction with a convincing IC50 (half-maximal inhibitory concentration) value of 13.07 µg/ml, which is considerable for developing therapeutic drug leads against human hepatoma. Reversed phase High-Performance Liquid Chromatography (HPLC) was finally conducted for the same column fraction and the result indicates the presence of one or several main cytotoxic compounds against human HepG2 cells.

Keywords: Cytotoxic compounds, half-maximal inhibitory concentration, high-performance liquid chromatography, human HepG2 cells, Nigella sativa seeds, Sulforhodamine-B assay-guided fractionation.

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Authors: U. Pangnakorn, S. Kanlaya, C. Kuntha

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Raw wood vinegar was purified by both standing and filtering methods. Toxicity tests were conducted under laboratory conditions by the topical application method (contact poison) and feeding method (stomach poison). Larvicidal activities of wood vinegar at four different concentrations (10, 15, 20, 25 and 30 %) were studied against second instar larvae of housefly (Musca domestica L.). Four replicates were maintained for all treatments and controls. Larval mortality was recorded up to 96 hours and compared with the larval survivability by two methods of larvicidal bioassay. Percent pupation and percent adult emergence were observed in treated M. domestica. The study revealed that the feeding method gave higher efficiency compared with the topical application method. Larval mortality increased with increasing concentration of wood vinegar and the duration of exposure. No mortality was found in treated M. domestica larvae at minimum 10% concentration of wood vinegar through the experiments. The treated larvae were maintained up to pupa and adult emergence. At 30% maximum concentration larval duration was extended to 11 days in M. domestica for topical application method and 9 days for feeding method. Similarly the pupal durations were also increased with increased concentrations (16 and 24 days for topical application method and feeding method respectively at 30% concentration) of the treatments.

Keywords: Housefly (Musca domestica L.), wood vinegar, mortality, topical application, feeding

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Authors: U. Pangnakorn, S. Kanlaya

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The efficiency of wood vinegar mixed with each individual of three plants extract such as: citronella grass (Cymbopogon nardus), neem seed (Azadirachta indica A. Juss), and yam bean seed (Pachyrhizus erosus Urb.) were tested against the second instar larvae of housefly (Musca domestica L.). Steam distillation was used for extraction of the citronella grass while neem and yam bean were simple extracted by fermentation with ethyl alcohol. Toxicity test was evaluated in laboratory based on two methods of larvicidal bioassay: topical application method (contact poison) and feeding method (stomach poison). Larval mortality was observed daily and larval survivability was recorded until the survived larvae developed to pupae and adults. The study resulted that treatment of wood vinegar mixed with citronella grass showed the highest larval mortality by topical application method (50.0%) and by feeding method (80.0%). However, treatment of mixed wood vinegar and neem seed showed the longest pupal duration to 25 day and 32 days for topical application method and feeding method respectively. Additional, larval duration on treated M. domestica larvae was extended to 13 days for topical application method and 11 days for feeding method. Thus, the feeding method gave higher efficiency compared with the topical application method.

Keywords: Housefly (Musca domestica L.), neem seed (Azadirachta indica), citronella grass (Cymbopogon nardus) yam bean seed (Pachyrhizus erosus), mortality.

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Authors: Samy A. Selim

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The antimicrobial, antiplasmid and cytotoxic activities of marine algae Halimeda opuntia and Sarconema filiforme were investigated. Antimicrobial bioassay against some human pathogenic bacteria and yeast were conducted using disc diffusion method. Halimeda extract exhibited antibacterial activity against six species of microrganisms, with significant inhibition against Staphylococcus aureus. While Sarconema extract was better potent as antifungal against Candida albicans. Comparative antibacterial studies showed that Halimeda extract showed equivalent or better activity as compared with commercial antibiotic when tested against Staphylococcus aureus. Further tests conducted using dilution method showed both extracts as having bacteriostatic mode of action against the tested microorganisms. Methanol extract of two species showed significant cytotoxicity (LC50 <500μg) on brine shrimp. Halimeda opuntia showed highest cytotoxic activity (LC50 =192.3μg). Also, the present investigation was undertaken to investigate the ability of methanolic extract of the algal extracts to cure R-plasmids from certain clinical E. coli isolates. The active fraction of Halimeda and Sarconema could cure plasmids from E. coli at curing efficiencies of approximately 78%. The active fraction mediated plasmid curing resulted in the subsequent loss of antibiotic resistance encoded in the plasmids as revealed by antibiotic resistance profile of cured strains. The screening results confirm the possible use of marine algae Halimeda opuntia and Sarconema filiforme as a source of pharmacological benefits.

Keywords: Antimicrobial, antiplasmid Cytotoxicity, Marine Algae.

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Authors: Hajji Lobna, Chattaoui Mayssa, Regaieg Hajer, M'Hamdi-Boughalleb Naima, Rhouma Ali, Horrigue-Raouani Najet

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In this study, three local isolates of Trichoderma (Tr1: T. viride, Tr2: T. harzianum and Tr3: T. asperellum) were isolated and evaluated for their biocontrol effectiveness under in vitro conditions and in greenhouse. In vitro bioassay revealed a biopotential control against Fusarium oxysporum f. sp. radicis lycopersici and Meloidogyne javanica (RKN) separately. All species of Trichoderma exhibited biocontrol performance and (Tr1) Trichoderma viride was the most efficient. In fact, growth rate inhibition of Fusarium oxysporum f. sp. radicis lycopersici (FORL) was reached 75.5% with Tr1. Parasitism rate of root-knot nematode was 60% for juveniles and 75% for eggs with the same one. Pots experiment results showed that Tr1 and Tr2, compared to chemical treatment, enhanced the plant growth and exhibited better antagonism against root-knot nematode and root-rot fungi separated or combined. All Trichoderma isolates revealed a bioprotection potential against Fusarium oxysporum f. sp. radicis lycopersici. When pathogen fungi inoculated alone, Fusarium wilt index and browning vascular rate were reduced significantly with Tr1 (0.91, 2.38%) and Tr2 (1.5, 5.5%), respectively. In the case of combined infection with Fusarium and nematode, the same isolate of Trichoderma Tr1 and Tr2 decreased Fusarium wilt index at 1.1 and 0.83 and reduced the browning vascular rate at 6.5% and 6%, respectively. Similarly, the isolate Tr1 and Tr2 caused maximum inhibition of nematode multiplication. Multiplication rate was declined at 4% with both isolates either tomato infected by nematode separately or concomitantly with Fusarium. The chemical treatment was moderate in activity against Meloidogyne javanica and Fusarium oxysporum f. sp. radicis lycopersici alone and combined.

Keywords: Trichoderma spp., Meloidogyne javanica, Fusarium oxysporum f.sp. radicis lycopersici, biocontrol.

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Authors: Katie Emerson, Sara Perez-Ojalvo, Jim Komorowski, Danielle Greenberg

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The purpose of this study was to evaluate arginase activity levels in response to combinations of an inositol-stabilized arginine silicate (ASI; Nitrosigine®), L-arginine, and Inositol. Arginine acts as a vasodilator that promotes increased blood flow resulting in enhanced delivery of oxygen and nutrients to the brain and other tissues. Arginase, found in human serum, catalyzes the conversion of arginine to ornithine and urea, completing the last step in the urea cycle. Decreasing arginase levels maintains arginine and results in increased nitric oxide production. This study aimed to determine the most effective combination of ASI, L-arginine and inositol for minimizing arginase levels and therefore maximize ASI’s effect on cognition. Serum was taken from untreated healthy donors by separation from clotted factors. Arginase activity of serum in the presence or absence of test products was determined (QuantiChrom™, DARG-100, Bioassay Systems, Hayward CA). The remaining ultra-filtrated serum units were harvested and used as the source for the arginase enzyme. ASI alone or combined with varied levels of Inositol were tested as follows: ASI + inositol at 0.25 g, 0.5 g, 0.75 g, or 1.00 g. L-arginine was also tested as a positive control. All tests elicited changes in arginase activity demonstrating the efficacy of the method used. Adding L-arginine to serum from untreated subjects, with or without inositol only had a mild effect. Adding inositol at all levels reduced arginase activity. Adding 0.5 g to the standardized amount of ASI led to the lowest amount of arginase activity as compared to the 0.25 g, 0.75 g or 1.00g doses of inositol or to L-arginine alone. The outcome of this study demonstrates an interaction of the pairing of inositol with ASI on the activity of the enzyme arginase. We found that neither the maximum nor minimum amount of inositol tested in this study led to maximal arginase inhibition. Since the inhibition of arginase activity is desirable for product formulations looking to maintain arginine levels, the most effective amount of inositol was deemed preferred. Subsequent studies suggest this moderate level of inositol in combination with ASI leads to cognitive improvements including reaction time, executive function, and concentration.

Keywords: Arginine, blood flow, colorimetry, urea cycle.

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Authors: K. J. Sannapapamma, H. Malligawad Lokanath, Sakeena Naikwadi

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Eco-friendly textiles are gaining importance among the consumers and textile manufacturers in the healthcare sector due to increased environmental pollution which leads to several health and environmental hazards. Hence, the research was designed to cultivate and develop the organic cotton knit, to prepare and characterize the Vetiver oil microcapsules for textile finishing and to access the wash durability of finished knits. The cotton SAHANA variety grown under organic production systems was processed and spun into 30 single yarn dyed with four natural colorants (Arecanut slurry, Eucalyptus leaves, Pomegranate rind and Indigo) and eco dyed yarn was further used for development of single jersy knitted fabric. Vetiveria zizanioides is an aromatic grass which is being traditionally used in medicine and perfumery. Vetiver essential oil was used for preparation of microcapsules by interfacial polymerization technique subjected to Gas Chromatography Mass Spectrometry (GCMS), Fourier Transform Infrared Spectroscopy (FTIR), Thermo Gravimetric Analyzer (TGA) and Scanning Electron Microscope (SEM) for characterization of microcapsules. The knitted fabric was finished with vetiver oil microcapsules by exhaust and pad dry cure methods. The finished organic knit was assessed for laundering on antimicrobial efficiency and aroma intensity. GCMS spectral analysis showed that, diethyl phthalate (28%) was the major compound found in vetiver oil followed by isoaromadendrene epoxide (7.72%), beta-vetivenene (6.92%), solavetivone (5.58%), aromadenderene, azulene and khusimol. Bioassay explained that, the vetiver oil and diluted vetiver oil possessed greater zone of inhibition against S. aureus and E. coli than the coconut oil. FTRI spectra of vetiver oil and microcapsules possessed similar peaks viz., C-H, C=C & C꞊O stretching and additionally oil microcapsules possessed the peak of 3331.24 cm^-1 at 91.14 transmittance was attributed to N-H stretches. TGA of oil microcapsules revealed that, there was a minimum weight loss (5.835%) recorded at 467.09°C compared to vetiver oil i.e., -3.026% at the temperature of 396.24°C. The shape of the microcapsules was regular and round, some were spherical in shape and few were rounded by small aggregates. Irrespective of methods of application, organic cotton knits finished with microcapsules by pad dry cure method showed maximum zone of inhibition compared to knits finished by exhaust method against S. aureus and E. coli. The antimicrobial activity of the finished samples was subjected to multiple washing which indicated that knits finished with pad dry cure method showed a zone of inhibition even after 20^th wash and better aroma retention compared to knits finished with the exhaust method of application. Further, the group of respondents rated that the 5^th washed samples had the greater aroma intensity in both the methods than the other samples. Thus, the vetiver microencapsulated organic cotton knits are free from hazardous chemicals and have multi-functional properties that can be suitable for medical and healthcare textiles.

Keywords: Exhaust and pad dry cure finishing, interfacial polymerization, organic cotton knits, vetiver oil microcapsules.

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