Brine Shrimp Larvae Research Articles

Sustainable PHBH-Alumina Nanowire Nanocomposites: Properties and Life Cycle Assessment.

Poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) (PHBH) is a bio-based polyester with the potential to replace some common polymers of fossil origin. However, PHBH presents serious limitations, such as low stiffness, tendency to undergo crystallization over long time periods and low resistance to thermal degradation during processing. In this work, we studied the use of alumina nanowires to generate PHBH-alumina nanocomposites, modifying the properties of PHBH to improve its usability. Solvent casting and melt blending were used to produce the nanocomposites. Then, their physicochemical properties and aquatic toxicity were measured. Finally, LCA was used to evaluate and compare the environmental impacts of several scenarios relevant to the processing and end of life (EoL) conditions of PHBHs. It was observed that, at low concentrations (3 wt.%), the alumina nanowires have a small positive impact on the stiffness and thermal degradation for the samples. However, for higher concentrations, the observed effects differed for each of the applied processing techniques (solvent casting or melt blending). The toxicity measurements showed that PHBH alone and in combination with alumina nanowires (10 wt.%) did not produce any impact on the survival of brine shrimp larvae after 24 and 48 h of exposure. The 18 impact categories evaluated by LCA allowed defining the most environmentally friendly conditions for the processing and EoL of PHBHs, and comparing the PHBH-related impacts to those of some of the most common fossil-based plastics. It was concluded that the preferable processing technique for PHBH is melt blending and that PHBH is unquestionably more environmentally friendly than every other analyzed plastic.

Indole analogues decreasing the virulence of Vibrio campbellii towards brine shrimp larvae.

Indole signalling has been proposed as a potential target for the development of novel virulence inhibitors to control bacterial infections. However, the major structural features of indole analogues that govern antivirulence activity remain unexplored. Therefore, we investigated the impact of 26 indole analogues on indole-regulated virulence phenotypes in Vibrio campbellii and on the virulence of the bacterium in a gnotobiotic brine shrimp model. The results demonstrated that 10 indole analogues significantly increased the fluorescence of indole reporter strain Vibrio cholerae S9149, 21 of them decreased the swimming motility of V. campbellii, and 13 of them significantly decreased the biofilm formation of V. campbellii. Further, we found that 1-methylindole, indene, 2,3-benzofuran, thianaphthene, indole-3-acetonitrile, methyl indole-3-carboxylate, 3-methylindole, and indole-2-carboxaldehyde exhibited a significant protective effect on brine shrimp larvae against V. campbellii infection, resulting in survival rates of challenged brine shrimp above 80%. The highest survival of shrimp larvae (98%) was obtained with indole-3-acetonitrile, even at a relatively low concentration of 20 μM. Importantly, the indole analogues did not affect bacterial growth, both in vitro and in vivo. These results indicate the potential of indole analogues in applications aiming at the protection of shrimp from vibriosis.

Evaluation of antioxidant, antimicrobial and antitumor compounds production potential of Streptomyces antioxidans PU-12M

Streptomycetes are prolific producers of biologically active secondary metabolites with great structural and functional diversity, which are widely used as pharmaceutical products. The strain Streptomyces sp. PU-12M was investigated for its potential to produce antioxidant, antimicrobial and antitumor compounds. For this purpose, the laboratory scale fermentation (10 liters working volume) of the strain was performed followed by cell separation, solvent extraction and partial purification of biologically active compounds by silica gel fractionation column chromatography and preparative TLC. The antioxidant activity was determined by DPPH radical scavenging assay, antimicrobial activity by agar well diffusion assay while in vitro antitumor activity was determined by MTT and SRB assays. Altogether 24 partially purified compounds or fractions were retrieved from the extract of strain Streptomyces Sp. PU-12M and were subsequently investigated for biological activity. Among them, three fractions were found to have potent antioxidant activity. The crude extracts and the partially purified fractions were found to have promising antimicrobial activity with MIC up to 3.2mg/ml. The partially purified fractions exhibited varied cytotoxicity ranging from 40% to 100% mortality against brine shrimp (Artemia salina) larvae and 22% to 67% growth inhibition of HCT 116 colorectal cancer cell lines. The study reveals that Streptomyces antioxidans PU-12M is a proliferent producer of bioactive secondary metabolites and can be exploited further to produce the commercially useful bioactive molecules.

Isolation and Characterization of Friedelin and 5-DodecylResorcinol from the Stem Bark Extract of Pterocarpus erinaceus

Pterocarpus erinaceus (Fabaceae) is a medicinal plant used in the treatment of several diseases such as, fever, diarrhea, and dysentery. The aim of this research work is to isolate phytochemicals, evaluate antimicrobial and cytotoxic activities of the extract/isolated compounds from the stem bark of Pterocarpus erinaceus. The plant material was air dried and extracted with a polarity gradient of petroleum ether, ethyl acetate, and methanol. Purification of the methanol extract using column chromatography yielded two compounds: friedelin (1) and 5-dodecylresorcinol (2). Antimicrobial screening of the extracts and isolated compounds was conducted using the disc diffusion method, while the cytotoxicity was tested on the extracts using Brine Shrimp larvae. The methanol extract was more toxic (LC50 of 0.93 µg/mL) than petroleum ether (PER-PE) and ethyl acetate (PER-EA) extracts. Antimicrobial screening of the plant extracts and compounds revealed a moderate activity at 1000 µg/mL.

Isolation and Cytotoxicity Evaluation of Long Chain Bioactive Compounds from Commiphora swynnertonii (Burtt)

Commiphora swynnertonii (Burtt) is a medicinal plant with diverse traditional uses in Tanzania. However, there is limited information on isolation and cytotoxicity properties of bioactive compounds responsible for its pharmacological activities. Hence, this study attempted to isolate and assess toxicity properties of bioactive compounds from its leaves, stem barks and whole root. Isolation of bioactive compounds was done by column chromatography method. There structures were deduced with the aid of NMR and GC-MS data and comparison with spectral data available in the literatures. Cytotoxicity evaluation was done by brine shrimp lethality test (BSLT) method. Chromatographic separation led to isolation of Hexacosane (1), Ethyl octadecanoate (2) and Octacosanol (3) from the leaves; Ethyl pentadecanoate (4) and Hexacosanoic acid (5) from the stem barks and Ethyl tetradecanoate (6), Methyl pentadecanoate (7) and Ethyl hexadecanoate (8) from the whole root. BSLT evaluation revealed that all compounds were non-toxic to brine shrimp larvae (LC50 >100 µg/ml). This study reports the isolation of these compounds for the first time from this plant, moreover, their presence demonstrates the usefulness of C. swynnertonii as a good source of pharmacological active reported compounds which authenticate its diverse uses in traditional medicine. Furthermore, the revealed non-toxic effect of these compoundsshows safety and possibly its traditional use has no cytotoxicity effect. Yet, further studies on toxicity of these compounds are recommended.

Halogenated Indoles Decrease the Virulence of Vibrio campbellii in a Gnotobiotic Brine Shrimp Model.

ABSTRACTIndole signaling is viewed as a potential target for antivirulence therapy against antibiotic-resistant pathogens because of its link with the production of virulence factors. This study examined the antimicrobial and antivirulence properties of 44 indoles toward Vibrio campbellii. Based on the results, 17 halogenated indole analogues were selected, as they significantly improved the survival of brine shrimp larvae challenged with V. campbellii. Specifically, 6-bromoindole, 7-bromoindole, 4-fluoroindole, 5-iodoindole, and 7-iodoindole showed a high protective effect, improving the survival of brine shrimp to over 80% even at a low concentration of 10 μM. To explore the impact of selected indole analogues on bacterial virulence phenotypes, swimming motility, biofilm formation, protease activity, and hemolytic activity of V. campbellii were determined. The results showed that all of the 17 selected indole analogues decreased swimming motility at both 10 μM and 100 μM. Most of the indole analogues decreased biofilm formation at a concentration of 100 μM. In contrast, only a slightly decreased protease activity and no effect on hemolytic activity were observed at both concentrations. To our knowledge, this is the first study of the structure-activity relation of halogenated indole analogues with respect to virulence inhibition of a pathogenic bacterium in an in vivo host model system, and the results demonstrate the potential of these compounds in applications aiming at the protection of shrimp from vibriosis, a major disease in aquaculture.IMPORTANCE Bacterial diseases are a major problem in the aquaculture industry. In order to counter this problem, farmers have been using antibiotics, and this has led to the evolution and spread of antibiotic resistance. In order for the aquaculture industry to further grow in a sustainable way, novel and sustainable methods to control diseases are needed. We previously reported that indole signaling is a valid target for the development of novel therapies to control disease caused by Vibrio campbellii and related bacteria, which are among the major bacterial pathogens in aquaculture. In the present study, we identified indole analogues that are more potent in protecting brine shrimp (a model organism for shrimp) from V. campbellii. To our knowledge, this is the first study of the structure-activity relation of halogenated indole analogues with respect to virulence inhibition of a pathogenic bacterium in an in vivo host model system.

Cytotoxicity and Nematicidal Potential of Leaf Extracts of <em>Adansonia digitata</em> and <em>Khaya senegalensi</em>s on Root Knot Nematode (<em>Meloidogyne incognita</em>) Associated with Cabbage

Purpose: Cabbage, an indispensable vegetable is often plagued by Meloidogyne incognita, a circumstance which lessens yield. Extracts of Khaya senegalensis and Adansonia digitata were appraised for possible nematicidal activity to check the menace of M. incognita sequel to the undesirable effect of synthetic nematicides in the environment. Research Method: Leaf materials were extracted in organic and aqueous extracts. The extracts were used as soil admix at 0, 150, 200 and 250 g/Kg soil, in a randomized complete block design experiment. Carbofuran a synthetic nematicide served as positive check for the extracts. Data was taken on vegetative growth, yield and nematode population. Lethality of the various extracts to brine shrimp larvae was also assessed. Findings: The aqueous extract was not as productive as the ethanol and methanol extracts. A. digitata extracts demonstrated weak nematicidal activity as against what was observed in cabbage plants treated with K. senegalensis extracts. Cytotoxicity evaluation of the organic and aqueous extracts of A. digitata indicated that the methanol extract expressed weak cytotoxicity (500 µg/ml - 1000 µg/ml), while the aqueous and ethanol extracts were not toxic to the brine shrimps (LC50 >1000µg/mL). K. senegalensis extracts were active against brine shrimp larvae with LC50 values <1000µg/mL The methanol and ethanol extracts demonstrated strong cytotoxicity with LC50 values <100µg/ml. Extracts from K. senegalensis significantly (p=0.05) increased the vegetative growth of cabbage plants. Originality/ Value: These results signify that the species have a probable use in the bio-pesticide industry, without any toxicity as the synthetics.

CYTOTOXICITY AND ANTIOXIDANT ACTIVITY OF Boerhavia adscendens Willd.

Boerhavia adscendens (B. adscendens) is a medicinal plant with widespread use in folk medicine. The present study was aimed at evaluation of the cytotoxicity and antioxidant activity of the crude ethanol extract of B. adscendens. The cytotoxicity of the extract was determined using brine shrimp lethality assay while its antioxidant activity was assessed using ferric reducing antioxidant potential (FRAP). The study revealed that the B. adscendens extract has moderate cytotoxic effect against the brine shrimp larvae with LC50 of 100 μg/mL relative to K2Cr2O4 standard with LC50 of 0.01 μg/mL. The FRAP determination for B. adscendens extract yielded an IC50 of 34.56 μg/mL while the ascorbic acid standard had IC50 value of 31.25 μg/mL. These results demonstrate that the B. adscendens ethanol extract has moderate cytotoxicity and quite a significant antioxidant activity. Consequently, these findings may partly explain the usefulness of B. adscendens in traditional medicine.

Structural Elucidation and Toxicity Evaluation of Bioactive Compounds from the Leaves and Stem woods of Synadenium glaucescens Pax.

The leaves and stem woods of Synadenium glaucescens Pax. are traditionally used for treatment of various human diseases in Tanzania. However, there is limited information on the structural elucidation and toxicity properties of bioactive compounds responsible for their pharmacological properties. Thus, this study was aimed to elucidate and evaluate toxicity properties of bioactive compounds from these parts. Separation of bioactive compounds was achieved by using column chromatographic method, whereas their structures were determined by GC-MS and NMR data and by comparing with literature spectroscopic data. Toxicity properties of isolated compounds was done by brine shrimp lethality test (BSLT) method. In this study, column chromatographic separation led to isolation of tetracosane (1), hexadecyl palmitate (2), octacosane (3), tetracosanol (4) and β-sitosterol (5) from the leaves and 1-hexacosene (6), campesterol (7), octadecanoic acid (8) and tetracosanoic acid (9) from the stem woods. BSLT of compounds isolated in sufficient amount (1, 2, 3, 4, 5 and 7) were revealed to be non-toxic to brine shrimp larvae (LC50 >100 µg/ml). Except compound 5, this study reports the isolation of other eight (8) compounds for the first time from this plant. Therefore, it can be reasonably concluded that S. glaucescens is a good source of bioactive compounds which justify its traditional uses in treatment of various ailments. Additionally, observed non-toxic effect of the tested compounds indicates safety and it is expected that its traditional use has no toxicity effect.

Phytochemical screening and Cytotoxicity assessment of Plantago lanceolata L. root extracts on Colorectal cancer cell lines and Brine shrimp larvae and determination of the median lethal dose in mice

Colorectal cancer is one of the most frequently diagnosed cancers, with a high mortality rate globally. Plantago lanceolata L. is a medicinal herb from the Plantaginaceae. P. lanceolata has various medicinal uses without significant side effects. The cytotoxic activity, biocompatibility, and in vivo toxicity of the methanolic and acetonic extracts of P. lanceolata root were evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, hemolysis, Artemia salina, and mice, respectively. The phytochemical content was performed by GC–MS. The root extract of P. lanceolata was cytotoxic, and IC50 values of methanolic and acetonic root extracts were 282.94, and 119.68 μg/mL on HCT-116 at 72 h, respectively. The hemolysis degree of the methanolic root extract was <1% at 400 μg/mL. The percentage of the lethality of the nauplii was <5% at 1000 μg/mL. One week after the oral administration with 2000 mg/kg, none of the mice died. According to the GC/MS analysis, P. lanceolata extracts contained potential cytotoxic compounds, such as stearic acid (4.90%) and Linoleic acid (3.04%) in methanolic extract and Bis(2-ethylhexyl) phthalate (21.04%) and 1,2- Benzenedicarboxylic acid, mono(2-ethylhexyl) ester (72.60%) in the P. lanceolata root acetonic extract. P. lanceolata methanolic extracts were identified as a biocompatible source. However, the IC50 of crude extracts has been obtained to be above the standard, according to the US National Cancer Institute. This study reported a good relationship between the in vitro and in vivo tests, and these tests are a useful tool for predicting and confirming toxicity in plant extracts.

Effect of phytosterols on osteoarthritis studied using insilico and CAM assays

Aim: The aim of the present study was to evaluate the anti-arthritic and anti-inflammatory effects of stigmasterol and β-sitosterol. Method: The CAM test was used to investigate the anti-arthritic and anti-inflammatory properties of stigmasterol and -sitosterol, which were obtained commercially. The anti-oxidant activity of the 2, 2'-Azino-Bis-3-Ethylbenzothiazoline-6-Sulfonic Acid (ABTS) and 2,2-diphenyl-1-picrylhydrazyl (DPPH) assays were measured in vitro. Using Autodock 4.2.6, in silico docking of plant sterol was performed to verify the binding effectiveness of the plantsterol with the receptor. The Chick chorioallantoic membrane (CAM) assay was studied for chondrogenesis in correlation to infiltration of inflammatory mediators in a lethality study of Brine Shrimp larvae in which the administration of test (Phytosterols) and standard (NaCl, NaOH) drugs was estimated. By estimating the toxicity of the phytosterols, the Chick chorioallantoic membrane (CAM) assay was studied for chondrogenesis. Results: The CAM experiment demonstrates that the mTORC1 signalling pathway has anti-arthritic and anti-inflammatory properties, as well as the ability to deactivate particular inflammatory mediators, resulting in increased cell recovery and joint health. In the CAM experiment in chick embryo, 10 g and 20 g of each phytosterol were used, respectively.

3D Acoustic Manipulation of Living Cells and Organisms Based On 2D Array.

Flexible manipulation techniques for living cells and organisms are extremely useful tools for fundamental biomedical and life science research. Acoustic tweezers, which permit non-contact, label-free manipulation, are particularly suited to micromanipulation tasks as they provide a large acoustic radiation force and can be applied in various media. Here, we describe the design and fabrication of a 3 MHz, 64-element (8 × 8), 2D planar ultrasound array that realizes the multidimensional translation, rotation, orientation, and levitation of living cells and organisms. The focusing vortex and twin fields are generated using the holographic acoustic elements framework method. We demonstrate that the eggs and larvae of brine shrimp can be translated along a preset trajectory by controlling the central position of the vortex. By multiplexing counterclockwise vortices, clockwise vortices, and twin trap fields in a time sequence, the rotation direction of the shrimp eggs can be switched in real time, while non-spherical larvae can be reoriented. Moreover, the reflection of the acoustic beam can lift eggs and larvae from the bottom of the culture dish and further manipulate them in the vertical and horizontal directions. Additionally, we present quantitative analyses of the shrimp-egg rotation frequency with respect to the focal depths, topological charges of the vortex, and excitation voltages. These results indicate that acoustic tweezers based on 2D matrix arrays can realize complex and selective manipulation of living cells and organisms, thereby demonstrating their value for advancing research in the fields of cell assembly, tissue engineering, and micro-robot driving.

Biosynthesis, characterization, antibacterial activities of manganese nanoparticles using Arcopilus globulus and their efficiency in degradation of bisphenol A

The present study aimed to synthesize manganese nanoparticles (Mn NPs) using endophytic fungi (Arcopilus globulus strain KNUEP5). The Arcopilus globulus extracts (AGE) mediated manganese nanoparticles (AGE-Mn NPs) were characterized using various materials characterization equipment, including UV–vis spectrophotometer, TEM-EDS, FTIR, and XRD. The characterization results showed an intense absorption peak at 324 nm in the UV spectrum, which indicates the successful synthesis of AGE-Mn NPs. AGE-Mn NPs were found to be spherical with a size of < 200 nm as indicated by TEM analysis. AGE-Mn NPs exhibited a zeta potential size of 130.2 ± 23.69 nm with a zeta potential of −8.72 ± 0.96 mV. FTIR analysis indicated the involvement of the various functional molecules in the AGE to the synthesis of the AGE-Mn NPs. Further, XRD results proved the crystalline structure of AGE-Mn NPs. The antibacterial and CLSM assays revealed that the AGE-Mn NPs significantly inhibited the bacterial pathogen (Escherichia coli). In addition, AGE-Mn NPs were very good at triggering the photocatalytic degradation of BPA, which made it less toxic to brine shrimp larvae.

Pharmacological evaluation of the hypoglycemic and anti- Alzheimer’s activities of aerial parts of Breynia distachia (Phyllanthaceae)

Purpose: To determine the cytotoxic, bronchorelaxant, spasmolytic, antidiabetic, α-glucosidase, acetylcholinesterase, butyrylcholinesterase, α-chymotrypsin and lipoxygenase inhibitory attributes of methanol and dichloromethane extracts of the aerial parts of Breynia distachia.Methods: The dichloromethane and methanol extracts of the aerial parts of the plant were prepared by maceration. Various ex vivo assays were employed, such as the brine shrimp lethality assay, lipoxygenase inhibitory activity assay, α-glucosidase inhibitory assay and α-chymotrypsin assay, as well as assays to assess the spasmolytic and bronchorelaxant activity. Meanwhile, the hypoglycaemic effect were analysed using an alloxan-induced diabetic model in Wistar albino rats.Results: The methanol extract (aerial) showed significant (p ≤ 0.05) cytotoxicity towards brine shrimp larvae at concentrations of 10, 100 and 1,000 μg/mL, respectively, whereas the dichloromethane extract (aerial) of the plant showed non-significant (p ≥ 0.05) results. The methanol extract (aerial parts) also demonstrated significant (p ≤ 0.05) α-glucosidase inhibitory activity and lipoxygenase inhibitory activity, with IC50 (half-maximal inhibitory concentration) values of 40.37 ± 5.29 μg/mL and 132.9 ± 0.33 μg/mL, respectively, while the dichloromethane extract exhibited significant (p ≤ 0.05) α-glucosidase inhibitory activity, with an IC50 value of 135.43 ± 8.29 μg/mL. An in vivo antidiabetic model showed that the administration of 150 and 300 mg/kg methanol extract of the aerial parts significantly (p ≤ 0.05) lowered the blood glucose level in alloxan-induced diabetic rats compared to control (treated with water).Conclusion: Data from different in vitro and in vivo models suggest that the methanol extract (aerial parts) of B. distachia shows significant cytotoxic, bronchorelaxant, spasmolytic, antidiabetic and anti-Alzheimer’s activity Hence, these findings validate the folkloric use of B. distachia and highlight the need to further explore its medicinal potential and the phytoconstituents responsible for its pharmacological actions.

Screening of the Toxicity of Polystyrene Nano- and Microplastics Alone and in Combination with Benzo(a)pyrene in Brine Shrimp Larvae and Zebrafish Embryos.

The occurrence of nanoplastics (NPs) and microplastics (MPs) in aquatic ecosystems and their capacity to sorb hydrophobic pollutants is nowadays an issue of great concern. This study aimed to assess the potential bioavailability and acute toxicity of polystyrene (PS) NPs (50 and 500 nm) and of MPs (4.5 µm), alone and with sorbed benzo(a)pyrene (B(a)P), in the embryo/larval stages of brine shrimps and zebrafish. Exposure to pristine plastics up to 50.1 mg PS/L did not cause significant impact on brine shrimp survival, while some treatments of plastics-B(a)P and all concentrations of B(a)P (0.1–10 mg/L) resulted acutely toxic. In zebrafish, only the highest concentrations of MPs-B(a)P and B(a)P caused a significant increase of malformation prevalence. Ingestion of NPs was observed by 24–48 h of exposure in the two organisms (from 0.069 to 6.87 mg PS/L). In brine shrimps, NPs were observed over the body surface and within the digestive tract, associated with feces. In zebrafish, NPs were localized in the eyes, yolk sac, and tail at 72 h, showing their capacity to translocate and spread into the embryo. MP ingestion was only demonstrated for brine shrimps. In zebrafish embryos exposed to plastics-B(a)P, B(a)P appeared in the yolk sac of the embryos. The presence of B(a)P was also noticeable in brine shrimps exposed to 500 nm NPs-B(a)P. In conclusion, NPs entered and spread into the zebrafish embryo and PS NPs, and MPs were successful vectors of B(a)P to brine shrimp and zebrafish embryos. Particle size played a significant role in explaining the toxicity of plastics–B(a)P. Our study provides support for the idea that plastics may pose a risk to aquatic organisms when combined with persistent organic pollutants such as B(a)P.

Antifouling coating based on biopolymers (PCL/ PLA) and bioactive extract from the sea cucumber Stichopus herrmanni

An important challenge to decrease the toxic effects of the common biocides in marine environments and to achieve suitable ecofriendly natural antifouling coatings is to find effective natural antifoulants and efficient biodegradable coatings. In this study, antifouling activities of nine bioactive extracts (non-polar to polar) from different organs of the sea cucumber Stichopus herrmanni were tested against five bacterial strains, barnacle and brine shrimp larvae. The ethyl acetate extract of the body wall showed the highest in-vitro antifouling activity including high antibacterial and anti-barnacle activities and low toxicity against the brine shrimp as non-target organism. Based on these results, 10 phr of the ethyl acetate extract from S.herrmanni was added to different coatings consisting of polycaprolactone (PCL)/polylactic acid (PLA) blends containing various compositions of PLA (0, 10, and 20 wt.%). Polyvinyl chloride panels were coated with the prepared antifouling coatings and immersed in seawater for three months. Panel coated with PCL 80% /PLA 20% containing 10 phr of the antifoulant (panel-5), showed the highest resistance against fouling settlement with fouling coverage of 41.66% (P < 0.05). In addition, the lowest fouling weight was measured in panel-5 as well (81.00 ± 9.85 g) (P < 0.05). These findings indicate the antibacterial and antifouling potential of semi-polar bioactive extracts from the S. herrmanni body wall as natural antifoulants, as well as the enhanced antifouling performance of PCL/the natural antifoulant coatings by adding PLA.Graphical

Evaluation of cytotoxic activity of Fomes fomentarius extracts against brine shrimp larvae and colon adenocarcinoma cell line (SW742)

Aim: Cancer is one of the leading worldwide public health problems. Natural product has provided a valuable source biological active compound for maintaining humans health even after years of introduction of synthetic drugs. Fomes fomentarius (F. fomentarius), a fungus belongs to Polyporaceae family, is widely used for different medicinal purposes. Considering inadequate reports on the fungus, cytotoxic effect of its extracts on brine shrimp larvae and colon adenocarcinoma cell line (SW742) was investigated in this study. Methods: The fruiting body of the fungus was chopped and extracted with water, ethanol, methanol, and ethanol-methanol (50-50) solvents. Regarding the results of brine shrimp lethality assay (BSLA), ethanol extract showed the highest toxicity, so subjected to fractionation with chloroform, ethyl acetate, and water. The fractions were tested for cytotoxic activity in human colon adenocarcinoma (SW742) cells by MTT assay. Ethyl acetate fraction exhibited more cytotoxic activity against SW742 cell line. Therefore, it was chosen to be purified using silica gel column chromatography. Subfractions of ethyl acetate were tested for their cytotoxicity by MTT assay. The content of these subfractions were also analyzed using gas chromatography-mass spectrometry (GC-MS). Results: According to the results, ethanol extract had the greatest potency in BSLA, and ethyl acetate fraction showed the highest cell cytotoxicity. Based on GC-MS chromatogram, palmitic acid was the compound with the largest quantity in the three selected subfractions of ethyl acetate. Conclusion: It was found that palmitic acid with other compounds in F. fomentarius had cytotoxic activity against colon adenocarcinoma cells. This feature may provide a biological basis for further clinical evaluation.

In-vitro and ex-vivo evaluation of plant sterol in osteoarthritis by CAM Assay

The aim of the study was to evaluate the pharmacological effect of stigmasterol and β-sitosterol using anti-inflammatory and anti-arthritic in-vitro and ex-vivo methods. Stigmasterol and β-sitosterol was obtained commercially and the CAM assay was done to determine the anti-arthritic and anti-inflammatory effects. The DPPH (2,2-diphenyl-1-picrylhydrazyl) and ABTS (2, 2'-Azino-Bis-3-Ethylbenzothiazoline-6-Sulfonic Acid) assay was performed for anti-oxidant by in-vitro method. An evaluation of toxicity was determined by the lethality study assessment of Brine Shrimp larvae in which the administration of test (Phytosterols) and standard (NaCl, NaOH) drugs was estimated, by estimating the toxicity of the phytosterols the Chick chorioallantoic membrane (CAM) assay was studied for chondrogenesis in correlation to infiltration of inflammatory mediators. The CAM assay shows the anti-arthritic activity and anti-inflammatory activity by mTORC1 signalling pathway and deactivates certain inflammatory mediators thereby increasing cell recovery and joint health. The CAM assay in chick embryo done by using 10 μg and 20μg of each phytosterol respectively. The toxicity LC50 measured at 87.7 μg/ml of phytosterols and 1.99 μg/ml for potassium dichromate (Std). At higher dose of 20 μg/ml the CAM membrane was observed in reducing the inflammation caused in cartilages of synovial joint. From the study, Stigmasterol and β-sitosterol managed the inflammation and an oxidative property plays a significant role in human health care system. Keywords: Osteoarthritis, Stigmasterol, ?-Sitosterol, Inflammation, Anti-Oxidant.

Polarity Directed Appraisal of Pharmacological Potential and HPLC-DAD Based Phytochemical Profiling of Polygonum glabrum Willd.

The present study was designed to evaluate polarity-dependent extraction efficiency and pharmacological profiling of Polygonum glabrum Willd. Crude extracts of leaves, roots, stems, and seeds, prepared from solvents of varying polarities, were subjected to phytochemical, antioxidant, antibacterial, antifungal, antidiabetic, and cytotoxicity assays. Maximum extraction yield (20.0% w/w) was observed in the case of an acetone:methanol (AC:M) root extract. Distilled water:methanol (W:M) leaves extract showed maximum phenolic contents. Maximum flavonoid content and free radical scavenging potential were found in methanolic (M) seed extract. HPLC-DAD quantification displayed the manifestation of substantial quantities of quercetin, rutin, gallic acid, quercetin, catechin, and kaempferol in various extracts. The highest ascorbic acid equivalent total antioxidant capacity and reducing power potential was found in distilled water roots and W:M leaf extracts, respectively. Chloroform (C) seeds extract produced a maximum zone of inhibition against Salmonella typhimurium. Promising protein kinase inhibition and antifungal activity against Mucor sp. were demonstrated by C leaf extract. AC:M leaves extract exhibited significant cytotoxic capability against brine shrimp larvae and α-amylase inhibition. Present results suggest that the nature of pharmacological responses depends upon the polarity of extraction solvents and parts of the plant used. P. glabrum can be considered as a potential candidate for the isolation of bioactive compounds with profound therapeutic importance.

Comparative Analysis of Antioxidant and Anticancer Activities of Fruiting Bodies, Mycelial Biomass and Culture Liquid of Omphalotus olearius OBCC 2503 (Agaricomycetes) from Turkey.

Up to now, the comparative analysis of the antioxidant and anticancer activities of the ethyl acetate extracts from fruiting body, mycelial biomass, and culture liquid of Omphalotus olearius are not reported. In this study, antioxidant activity of different samples was compared by six different methods: DPPH and ABTS radical scavenging activities, metal chelating activity, reducing power, inhibition of β-carotene bleaching, and lipid peroxidation through ferric thiocyanate assay. Total phenolic contents of the samples were also determined. The mycelial biomass produced in vitro conditions presented the highest activity by five of them which was following by culture liquid. The potential antiproliferative effects of the extracts in cell culture, human cancer (A549, Caco2 and MCF-7) and healthy (CCD-19LU and CCD 841 CoN) cell lines by WST-1 assay was also evaluated. The highest anticancer effects were determined on A549 lung cancer cells treated with mycelial biomass extract and Caco2 colon cancer cells treated with culture liquid extract. The culture liquid was more effective on the cells than mycelial biomass according to IC50 values. All tested extracts were also evaluated for their toxicity against brine shrimp larvae (Artemia salina). O. olearius culture liquid and mycelial biomass extracts showed highest and lowest toxicity against A. salina, respectively.