Marine Extracts Research Articles

Green Inhibitor as Antibacteria and Antiscaling in Reverse Osmosis Desalination Plants

Today, reverse osmosis membranes are the leading technology for new desalination installations, however, a challenge facing widespread application of RO technology is membrane fouling. In the present study, we used an environmentally friendly green inhibitor as anti-scaling and anti-biofouling in reverse osmosis (RO) desalination plants. The influence of Sargassum sp., Corallina mediterranea, and Avicennia marina on RO membrane mineral scaling was evaluated using gypsum as a model scalant. Antibacterial properties for three marine extracts from Sargassum sp., C. mediterranea, and Avicennia marina were investigated with Gram-positive bacteria (ArthrobactersulfureusYACS14, Staphylococcus aureus) and Gram-negative bacteria (VibrioanguillarumMVM425, Escherichia coli). The antimicrobial results were detected for the two selected extracts as the most potent extracts (ethyl acetate, methanol crude extracts of the Avicennia marina leaves). Data showed that ratios of 3 and 5% recorded the highest suppression percentages (100%) for all tested bacteria including bacterial community collected from Eastern Harbor. On the other side, data confirmed that the anti-scalant properties by 100 ppm of Avicennia marina leave extract giving 85% of scale inhibition. The effect of Avicennia marina leaves extract for calcium sulfate dihydrate (gypsum) scaling on selected reverse osmosis (RO) membrane surfaces was investigated. The effect of different concentrations of Avicennia marina leaves extract was observed in the extent of surface scale coverage and surface crystal size among the membrane studied.

Ultrastructure-based Insights on Anti-Trichomonas vaginalis Effects of Selected Egyptian Red Sea Marine Resources

Background:Metronidazole is used for the treatment of trichomoniasis. However, a growing number ofTrichomonas vaginalis(T. vaginalis) isolates are now resistant, which is an urgent issue to search for new alternatives. Worldwide marine pharmacy confirms the enormous potential of sea species as a source of novel pharmaceuticals.Objective:This study aimed to investigate the anti-T. vaginalisactivities of ethanolic extracts of Red Sea marine resources, soft corals;Sarcophyton glaucumandLitophyton arboreumand methanolic extracts of Red Sea brown algae;Liagora farinosa, Colpomenia sinuosa, Hydroclathrus clathratus,andSargassum graminifolium, as well as sea cucumber (Holothuria fuscocinerea) and sea urchin (Echinometra mathaei).T. vaginalisgrowth inhibition was determined using 2 concentrations for each marine extract 10 and 100 µg/ml in comparison to media control. Drugs that showed good initial activity were further tested to calculate their IC50 in comparison to metronidazole. The ultrastructural impact of the more effective extracts was further assessed.Results:H. clathratus,L. farinose, sea urchinE. mathaeiand sea cucumberH. fuscocinereareduced the growth ofT. vaginaliseffectively and showed high activity with IC50 of 0.985±0.08, 0.949±0.04, 0.845±0.09 and 0.798±µg/ml±SD, respectively. Concerning microscopic analysis, marine extract and metronidazole-treated cells presented similar morphological changes. The nuclear membrane was damaged, the nuclei were dissolved, the rough endoplasmic reticulum was widened, and the chromatin was accumulated. In the cytoplasm, numerous autophagic vacuoles appeared, the organelles were disintegrated, the flagella were internalized and hydrogenosomes with altered morphologies were observed. The cell membrane was partially damaged, with cytoplasmic leakage and cell disintegration.Conclusion:This study describes the report on the activity and morphological changes induced by Egyptian Red Sea marine resources againstT. vaginalis. The results obtained herein presented new opportunitiess. Further, bio-guided fractionation and isolation of active compounds are needed.

Integrated Genomic and Metabolomic Approach to the Discovery of Potential Anti-Quorum Sensing Natural Products from Microbes Associated with Marine Samples from Singapore.

With 70% of the Earth’s surface covered in water, the marine ecosystem offers immense opportunities for drug discovery and development. Due to the decreasing rate of novel natural product discovery from terrestrial sources in recent years, many researchers are beginning to look seaward for breakthroughs in new therapeutic agents. As part of an ongoing marine drug discovery programme in Singapore, an integrated approach of combining metabolomic and genomic techniques were initiated for uncovering novel anti-quorum sensing molecules from bacteria associated with subtidal samples collected in the Singapore Strait. Based on the culture-dependent method, a total of 102 marine bacteria strains were isolated and the identities of selected strains were established based on their 16S rRNA gene sequences. About 5% of the marine bacterial organic extracts showed quorum sensing inhibitory (QSI) activity in a dose-dependent manner based on the Pseudomonas aeruginosa QS reporter system. In addition, the extracts were subjected to mass spectrometry-based molecular networking and the genome of selected strains were analysed for known as well as new biosynthetic gene clusters. This study revealed that using integrated techniques, coupled with biological assays, can provide an effective and rapid prioritization of marine bacterial strains for downstream large-scale culturing for the purpose of isolation and structural elucidation of novel bioactive compounds.

Antioxidant, Antiglycation and in vivo Antiaging Effects of Metabolite Extracts from Marine Sponge-associated Bacteria

In vitro antioxidant, antiglycation activities of marine sponge-associated bacterial extracts and potential antiaging properties on the model organism, fission yeast (Schizosaccharomyces pombe) were investigated in the present study. Spectrometry method was used to assess the chemical contents of the extracts. Antioxidant activity was measured using 2,2-diphenyl-1-picrylhydrazyl and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) radicals, while antiglycation was a fluorometric assay. Antiaging activity was tested using the spot test and chronological life span assays. In addition, mitochondrial activity, intracellular reactive oxygen species level and cell cycle analysis of Schizosaccharomyces pombe were evaluated to understand fully the effect of the selected extract. Highest amount of phenolic compounds was found in HAL-13 extract of 215.15 mg gallic acid equivalents/g extract. PTR-21 extract has the highest flavonoid and tannin contents of 6.42 mg quercetin equivalents/g extract and 5.88 mg catechin/g extract, respectively. The best antioxidant activity was found in HAL-08 extract with IC50 value of 17.10 and 59.39 µg/ml in the two antioxidant assays employed, while the highest activity of antiglycation was reported in the PTR-21 extract with IC50 value of 157.50 µg/ml. PTR-21 (500 µg/ml) extract exhibited antiaging properties in Schizosaccharomyces pombe. Indeed, PTR-21 extract could extend G1 phase of the cell division from 26 to 40 %. Interestingly, the corresponding extract could enhance the stress tolerance of phenotype in Schizosaccharomyces pombe against H2O2-induced oxidative stress. Such phenotypes might be induced by mitochondrial-reactive oxygen species adaptive signaling, as the extract application significantly enhanced yeast mitochondrial respiratory activity.

Free fatty acid profiling in marine algae extract by LC-MS/MS and isolation as well as quantification of the ω-3 fatty acid hexadeca-4,7,10,13-tetraenoic acid.

Undaria pinnatifida (Wakame) alga contains high amounts of hexadeca-4Z,7Z,10Z,13Z-tetraenoic acid which was reported to decrease the efficiency of cisplatin chemotherapeutics. To obtain a fatty acid enriched extract of this ω-3 poly-unsaturated fatty acid as an analytical standard, Wakame was used as source material for its extraction. A two-step extraction protocol consisting of a liquid-liquid extraction followed by solid-phase extraction with 3-aminopropyl silica in accordance to a normal-phase elution mode was developed. An ultra high performance liquid chromatography with electrospray ionization tandem mass spectrometry method based on sequential windowed acquisition of all theoretical fragment ion mass spectra allowed a simultaneous comprehensive group selective fatty acids profiling in untargeted manner and quantitative analysis of the targeted fatty acid. Hexadeca-4Z,7Z,10Z,13Z-tetraenoic acid was identified using high-resolution product ion spectra. The quantitative method was based on d5-deuterated hexadeca-4Z,7Z,10Z,13Z-tetraenoic acid which was employed as surrogate calibrant. Preliminary method validation was performed by evaluating detection and quantification limits, linear range, intra-assay and inter-day precision. Finally, a concentration of 421.2±14.9ng/mL (4% CV) of hexadeca-4Z,7Z,10Z,13Z-tetraenoic acid was determined in the extract which was further used as analytical standard.

New Mammalian Target of Rapamycin (mTOR) Modulators Derived from Natural Product Databases and Marine Extracts by Using Molecular Docking Techniques.

Mammalian target of rapamycin (mTOR) is a PI3K-related serine/threonine protein kinase that functions as a master regulator of cellular growth and metabolism, in response to nutrient and hormonal stimuli. mTOR functions in two distinct complexes—mTORC1 is sensitive to rapamycin, while, mTORC2 is insensitive to this drug. Deregulation of mTOR’s enzymatic activity has roles in cancer, obesity, and aging. Rapamycin and its chemical derivatives are the only drugs that inhibit the hyperactivity of mTOR, but numerous side effects have been described due to its therapeutic use. The purpose of this study was to identify new compounds of natural origin that can lead to drugs with fewer side effects. We have used computational techniques (molecular docking and calculated ADMET (Absorption, Distribution, Metabolism, Excretion, and Toxicity) parameters) that have enabled the selection of candidate compounds, derived from marine natural products, SuperNatural II, and ZINC natural products, for inhibitors targeting, both, the ATP and the rapamycin binding sites of mTOR. We have shown experimental evidence of the inhibitory activity of eleven selected compounds against mTOR. We have also discovered the inhibitory activity of a new marine extract against this enzyme. The results have been discussed concerning the necessity to identify new molecules for therapeutic use, especially against aging, and with fewer side effects.

PO-049 Quantitative proteomics unveils key mitotic regulators involved in the transition to androgen independent prostate cancer

IntroductionMarine invertebrate organisms have received great attention from the scientific community due to its richness in new bioactive compounds with potential anti-proliferative and anticancer activities. In the present study, the cytotoxic efficacy of four marine invertebrate-extracts against human adenocarcinoma colon cancer cells was investigated.Material and methodsHT29, SW480 and HGUE-C-1 human colon adenocarcinoma cell lines were cultured and exposed to 4 marine invertebrate extracts at different concentrations for 24 hour. The anti-proliferative properties were examined by MTT assay. Analysis of apoptosis and cell cycle stages were performed with the Muse Cell Analyzer (Millipore, Hayward, CA, USA). Mitochondrial functionality was determined by the ratio of MitoTracker Red CMXRos and MitoTracker Green FM fluorescent probes (Molecular Probes). Measurement of lactate dehydrogenase (LDH) was performance by a colorimetric assay (Roche Diagnostic, Germany).Results and discussionsThe four marine invertebrate extracts exhibited pronounced cytotoxic effects in the three cancer cell lines used. In this study, CR, PS, NA and NB extracts were able to modify the cell cycle of HGUE-C-1, HT-29 and SW-480 cells, by increasing cell population in the subG1 and G2/M phase in a dose-dependent manner. Results confirmed that NA, NB and CR extracts induced an increase in the percentage of cell population in early apoptosis by Annexin V staining. PS extract on the contrary, raised the late apoptotic population at different concentrations. Mitochondrial membrane depolarization rate, as indicator of apoptosis, was induced significantly by CR, NA and NB extracts in the three cell models used. In contrast PS reduced this parameter in a dose-dependent manner.Finally, the release of the enzyme lactate dehydrogenase (LDH) confirmed a cell death by a necrotic pathway for PS extract.ConclusionOur results strongly suggest that NB, NA and CR extracts exhibit antiproliferative and pro-apoptotic properties in colon cancer cell lines concomitantly to a decreased mitochondrial membrane potential. In contrast, PS extract shows an antiproliferative effect mediated by necrosis in the same cell models. This preliminary study suggests that these extracts present pharmacological potential. Further investigations to determine the responsible compounds and the underlying mechanism are currently ongoing.

English

Rhizoctonia solani is the causative agent of rice sheath blight, which has become a major problem in rice production. Seaweed provides a rich source of structurally diverse and biologically active secondary metabolite and is proved to be better in decreasing foliar fungal diseases which ultimately increase its fertility and help in the growth of plants. The use of natural products is the ultimate way of combating this disease. In this context, five different seaweeds such as Sargassum wightii, Dictyota bartyrensiana, Ulva reticulate, Gelidiella acerosa and Odonus niger were used together with the fish powder extract in the control of sheath blight disease in rice were studied. Evaluation of marine products against R. solani was carried out by paper disc assay; agar well method and mycelial dry weight. Among the five marine extracts tested, extracts of S. wightii [brown seaweed algae] at a high concentration (20%) was found to be the best in the reduction of spore germination (19.60%). The leaf extracts of S. wightii [brown seaweed algae] at highest concentration of 20% showed a maximum reduction in both paper disc method and agar well method with 44.65 and 45.90% zone of inhibition, respectively. The antifungal compounds were identified through gas chromatography mass spectroscopy. The results revealed that, 18 compounds were present in S. wightii and among that n-hexadecanoic acid which was closely related to 9, 12-octadecadienoic acid may be responsible for the inhibition of the growth of R. solani. The present study revealed that the efficacy of seaweed extracts against fungal pathogens may be due to higher levels and early accumulation of phenolics and phytoalexins, and the pot study proved that R. solani can be controlled by the application of marine products which may be further used for field study. Key words: Seaweeds, Rhizoctonia solani, antifungal compounds, rice.

Natural Products And Traditional Medicines For The Treatment Of Multidrug Resistant Bacteria

Multidrug resistant microorganisms (MDROs) have been responsible for numerous outbreaks of infectious epidemics, and pose a serious threat to global health. The lack of effective therapies for these MDROs, coupled with a reduced number of antimicrobial drugs in the pharmaceutical pipeline to treat these infections indicates an urgent need for research in this area. Of the resistant Gram-positive bacteria, methicillin-resistant Staphylococcus aureus (MRSA), multidrug-resistant Pseudomonas aeruginosa and multidrug-resistant tuberculosis (MDR-TB) continue to be the most problematic, but more recently there has been increasing reports of vancomycin-resistant Staphylococcus aureus (VRSA) infections. For highly-resistant Gram-negative bacteria, Vancomycin-resistant Enterococci (VRE), multidrug-resistant carbapenemase-producing Klebsiella pneumoniae and MDR-Acinetobacter baumannii are important as these bacteria are often only susceptible to older antimicrobial agents, such as the polymyxins that have a higher adverse event profile. Thus, there is an urgent need for new antibiotics and combinations of drugs to treat these emerging MDR bacteria. The lack of interest by pharmaceutical companies in the research and development of new antibiotics has increased research in the field of natural products. An increasing body of scientific evidence suggests that specific marine, fungal and medicinal plant extracts, as well as traditional medicine formulas and purified natural products are active against specific MDR-bacteria, and thus could be further investigated and possibly developed as clinical agents. This review focuses on recently published data for natural products active against MDR bacteria and evaluates some of the recent published in vitro data.

Protective Effects of Fucoidan on Epithelial-Mesenchymal Transition of Retinal Pigment Epithelial Cells and Progression of Proliferative Vitreoretinopathy

Background/Aims: Proliferative vitreoretinopathy (PVR) is a severe blinding complication of rhegmatogenous retinal detachment. Epithelial-mesenchymal transition (EMT) of retinal pigment epithelial (RPE) cells is thought to play a pivotal role in the pathogenesis of PVR. Fucoidan, a marine extract, reportedly has many benefits effects in a variety of tissues and organs such as anti-inflammation, anti-oxidative stress, and anti-carcinogenesis. In this study, we investigated the potential role of fucoidan on EMT in RPE cells and its effect on the development of PVR. Methods: MTS, Transwell, and collagen gel contraction assays were employed to measure the viability, migration, and contraction of RPE cells, respectively. mRNA and protein expression were evaluated via real-time quantitative PCR and western blot analysis, respectively. In vivo, a pigmented rabbit model of PVR was established to examine the anti-PVR effect of fucoidan. Results: Fucoidan reversed the transforming growth factor (TGF)-β1-induced EMT of RPE cells, including the increased expression of α-smooth muscle actin (α-SMA) and fibronectin and down-regulation of E-cadherin in human primary RPE cells. Moreover, the upregulation of phosphorylated Smad2/3 induced by TGF-β1 was suppressed by fucoidan. Fucoidan also inhibited the migration and contraction of RPE cells induced by TGF-β1. In vivo, fucoidan inhibited the progression of experimental PVR in rabbit eyes. Histological findings showed that fucoidan suppressed the formation of α-SMA-positive epiretinal membranes. Conclusion: Our findings regarding the protective effects of fucoidan on the EMT of RPE cells and experimental PVR suggest the potential clinical application of fucoidan as an anti-PVR agent.

Polyunsaturated ω-3 and ω-6 fatty acids, total carotenoids and antioxidant activity of three marine microalgae extracts obtained by supercritical CO2 and subcritical n-butane

Microalgae are one of the most promising sources for new functional food products, due to their ability to synthesize polyunsaturated fatty acids, pigments and natural antioxidants (e.g. carotenoids and phenolic compounds). The aim of this study was to compare the fatty acid composition, total carotenoids and antioxidant activity of Phaeodactylum tricornutum, Nannochloropsis oculata, and Porphyridium cruentum extracts obtained from supercritical CO2 (SC-CO2) and subcritical n-butane extraction methods SC-CO2 method was selective in extract saturated fatty acids (SFAs) for all microalgae species studied. On the other hand, subcritical n-butane was shown to be an excellent and powerful method to extract polyunsaturated fatty acids (PUFAs ω3+ω6). Regarding to total carotenoids and antioxidant activity using DPPH scavenging assay a significant correlation coefficient (R2=0.80) was found among the extracts independent of the extraction method tested. This indicates that carotenoid compounds might be the major contributor to the antioxidant capacities of these microalgae.

Marine algae extract attenuated osteoporosis in OVX mice, enhanced osteogenesis on human mesenchymal stem cells and promoted OPG expression

Osteoporosis is an aging disease that weakens bones and increases risks of sudden and unexpected fractures. Effects of Marine Algae Extract (MAE) on bone formation and resorption remain unknown. We investigated whether MAE could enhance anti-osteoporosis and induce bone formation by using ovariectomized (OVX) mouse model and cultured human bone marrow derived mesenchymal stem cells (MSCs). With calcium content and other mineral elements, MAE attenuated osteoporosis by increasing secretions of osteocalcin and osteoprotegerin (OPG) in serum in OVX mice and resulted in the increasing level of the ratio of OPG to RANKL (P < .05, n = 3–5). MAE treatment 800 mg/kg also can reduce the gain of body weight in OVX mice about 10%. In vitro, MAE enhanced human MSCs osteogenic differentiation and inhibited adipogenesis via upregulation of transcriptional level of OPG (Day 4, P < .05, n = 3; Day 7, P < .001, n = 3). In conclusion, MAE has potential to prevent osteoporosis in vivo and in vitro.

P 216 - The antiproliferative effects of four marine invertebrate ectracts in colon cancer cells in relation to the modulation of oxidative stress-related pathways

The goal of this study was to correlate the anti-proliferative activity of four marine invertebrate extracts on different colon cancer cell lines with free radical generation and modulation of oxidative stress-related pathways. Three human colorectal carcinoma cell lines (HGUE, HT29 and SW480) were treated with four marine extracts (10-100 µg/mL) for 24 h at 37 °C. Cell viability (IC50 values)was determined by MTT. Reactive oxygen species (ROS) production was measured with DCF-DA. Mitochondrial functionality was tested by MitoTracker Red and MitoTracker Green fluorescent probes. Marine extracts reduced cell viability of colon cancer cells starting at 10 µg/mL after 24 h the exposition. From 4 tested extracts, several exhibited pronounced cytotoxic effect, at least in one of the cell lines. The inhibition of cell proliferation was correlated to an increase of intracellular ROS content and a decrease of the mitochondrial membrane potential. Our results suggest that the cytotoxic properties of the tested extracts observed in this preliminary study are due to the presence of pro-oxidant activity compounds which are responsible of the anti-proliferative effect in colon cancer cell lines. These extracts present pharmacological potential and further investigations to determine which are responsible compounds and the mechanism are currently ongoing.

Ultrasensitive and high-throughput analysis of chlorophyll a in marine phytoplankton extracts using a fluorescence microplate reader.

Chlorophyll a (Chl a) is the predominant pigment in every single photosynthesizing organism including phytoplankton and one of the most commonly measured water quality parameters. Various methods are available for Chl a analysis, but the majority of them are of limited throughput and require considerable effort and time from the operator. The present study describes a high-throughput, microplate-based fluorometric assay for rapid quantification of Chl a in phytoplankton extracts. Microplate sealing combined with ice cooling was proved an effective means for diminishing solvent evaporation during sample loading and minimized the analytical errors involved in Chl a measurements with a fluorescence microplate reader. A set of operating parameters (settling time, detector gain, sample volume) were also optimized to further improve the intensity and reproducibility of Chl a fluorescence signal. A quadratic regression model provided the best fit (r 2=0.9998) across the entire calibration range (0.05-240pgμL-1). The method offered excellent intra- and interday precision (% RSD 2.2 to 11.2%) and accuracy (% relative error -3.8 to 13.8%), while it presented particularly low limits of detection (0.044pgμL-1) and quantification (0.132pgμL-1). The present assay was successfully applied on marine phytoplankton extracts, and the overall results were consistent (average % relative error -14.8%) with Chl a concentrations (including divinyl Chl a) measured by high-performance liquid chromatography (HPLC). More importantly, the microplate-based method allowed the analysis of 96 samples/standards within a few minutes, instead of hours or days, when using a traditional cuvette-based fluorometer or an HPLC system. Graphical abstract TChl a concentrations (i.e. sum of Chl a and divinyl Chl a in ng L-1) measured in seawater samples by HPLC and fluorescence microplate reader.

Marine actinomycete crude extracts with potent TRAIL-resistance overcoming activity against breast cancer cells.

Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) is a promising anticancer agent, as it can kill tumor cells selectively. In our search of bioactive natural products to overcome TRAIL-resistance, we isolated 47 actinomycete strains from different sediments and seawater samples collected from the Red Sea coast in Egypt and found four crude extracts (EGY1, EGY3, EGY24 and EGY34) displaying TRAIL sensitizing activity in the resistant breast cancer cell line MDA-MB-231. None of these crude extracts exhibited cytotoxic effect on normal mouse embryonic fibroblasts (MEF), with the exception of EGY34. Analysis of the signaling pathways underlying the sensitization of MDA-MB-231 cells to TRAIL-induced apoptosis, by western blotting, revealed that all crude extracts facilitated initiator caspase‑8/-10 activation upon TRAIL stimulation, but that in addition, EGY3 and EGY34, alone, induced strong ER-stress activation, with the appearance of BiP in the cytosolic extracts. Our results pave the way to the discovery and the development of marine-derived drugs for cancer therapy.

Small-Molecule Inhibitors of the SOX18 Transcription Factor

Pharmacological modulation of transcription factors (TFs) has only met little success over the past four decades. This is mostly due to standard drug discovery approaches centered on blocking protein/DNA binding or interfering with post-translational modifications. Recent advances in the field of TF biology have revealed a central role of protein-protein interaction in their mode of action. In an attempt to modulate the activity of SOX18 TF, a known regulator of vascular growth in development and disease, wescreened a marine extract library for potential small-molecule inhibitors. We identified two compounds, which inspired a series of synthetic SOX18 inhibitors, able to interfere with the SOX18 HMG DNA-binding domain, and to disrupt HMG-dependent protein-protein interaction with RBPJ. These compounds also perturbed SOX18 transcriptional activity in a cell-based reporter gene system. This approach may prove useful in developing a new class of anti-angiogenic compounds based on the inhibition of TF activity.

Potential role of marine algae extract on 3T3-L1 cell proliferation and differentiation: an in vitro approach.

BackgroundFrom ancient times, marine algae have emerged as alternative medicine and foods, contains the rich source of natural products like proteins, vitamins, and secondary metabolites, especially Chlorella vulgaris (C. vulgaris) contains numerous anti-inflammatory, antioxidants and wound healing substances. Type 2 diabetes mellitus is closely associated with adipogenesis and their factors. Hence, we aimed to investigate the chemical constituents and adipogenic modulatory properties of C. vulgaris in 3T3-L1 pre-adipocytes.ResultsWe analysed chemical constituents in ethanolic extract of C. vulgaris (EECV) by LC–MS. Results revealed that the EECV contains few triterpenoids and saponin compounds. Further, the effect of EECV on lipid accumulation along with genes and proteins expressions which are associated with adipogenesis and lipogenesis were evaluated using oil red O staining, qPCR and western blot techniques. The data indicated that that EECV treatment increased differentiation and lipid accumulation in 3T3-L1 cells, which indicates positive regulation of adipogenic and lipogenic activity. These increases were associated with up-regulation of PPAR-γ2, C/EBP-α, adiponectin, FAS, and leptin mRNA and protein expressions. Also, EECV treatments increased the concentration of glycerol releases as compared with control cells. Troglitazone is a PPAR-γ agonist that stimulates the PPAR-γ2, adiponectin, and GLUT-4 expressions. Similarly, EECV treatments significantly upregulated PPAR-γ2, adiponectin, GLUT-4 expressions and glucose utilization. Further, EECV treatment decreased AMPK-α expression as compared with control and metformin treated cells.ConclusionThe present research findings confirmed that the EECV effectively modulates the lipid accumulation and differentiation in 3T3-L1 cells through AMPK-α mediated signalling pathway.

Immunotoxicity of skin acid secretion produced by the sea slug Berthellina citrina in mice spleen: Histological and Immunohistochemical study

Acid secretion containing sulfuric and hydrochloric acids is a fascinating defensive phenomenon within many groups of marine organisms. This study aimed to investigate the mice spleen histology and immunotoxicity using skin acid secretion (SAS) of the sea slug Berthellina citrina after oral administration. The spleen showed atrophy in the white pulp, decrease in the splenocytes density, megakaryocytes cytoplasmic degeneration as well as inflammatory cells infiltrations. The white and red pulp splenocytes number decreased time-dependently in the treated spleens. Additionally, the size of the megakaryocytes increased as compared with the control. The administration with SAS increased the number of the IgA+ cells aggregation in the splenic red pulp. Furthermore, after 7days of the administration, large number of dispersed IgA+ cells were distributed in splenic parenchyma. The IgA+ cells numbers increased time-dependently as compared with those in the control. The aggregation sizes and number of the F4/80+ cell in the splenic red pulp were increased. Furthermore the F4/80+ cells numbers increased time-dependently as compared with those in the control. The UEAI+ cells were found as free cells but not in aggregations in the control splenic red pulp. Contradictory to the number of IgA+ cells and F4/80+ cells the number of the UEAI+ cells decreased time-dependently after administration with SAS. Hematologically, abnormal numbers of WBCs different cells were observed after administration with SAS. This study provides new insight about the toxicity of a marine extract may be used in natural products industry or medical applications.

Development and Application of a Novel SPE-Method for Bioassay-Guided Fractionation of Marine Extracts.

The biological diversity of marine habitats is a unique source of chemical compounds with potential use as pharmaceuticals, cosmetics and dietary supplements. However, biological screening and chemical analysis of marine extracts pose specific technical constraints and require adequate sample preparation. Here we report an improved method on Solid Phase Extraction (SPE) to fractionate organic extracts containing high concentration of salt that hampers the recovery of secondary metabolites. The procedure uses a water suspension to load the extracts on a poly(styrene-divynylbenzene)-based support and a stepwise organic solvent elution to effectively desalt and fractionate the organic components. The novel protocol has been tested on MeOH-soluble material from three model organisms (Reniera sarai, Dendrilla membranosa and Amphidinium carterae) and was validated on a small panel of 47 marine samples, including sponges and protists, within discovery programs for identification of immuno-stimulatory and anti-infective natural products.

Doxorubicin-conjugated magnetic iron oxide nanoparticles for pH-sensitive and magnetic responsive drug delivery

Acid secretion containing sulfuric and hydrochloric acids is a fascinating defensive phenomenon within many groups of marine organisms. This study aimed to investigate the mice spleen histology and immunotoxicity using skin acid secretion (SAS) of the sea slug Berthellina citrina after oral administration. The spleen showed atrophy in the white pulp, decrease in the splenocytes density, megakaryocytes cytoplasmic degeneration as well as inflammatory cells infiltrations. The white and red pulp splenocytes number decreased time-dependently in the treated spleens. Additionally, the size of the megakaryocytes increased as compared with the control. The administration with SAS increased the number of the IgA+ cells aggregation in the splenic red pulp. Furthermore, after 7 days of the administration, large number of dispersed IgA+ cells were distributed in splenic parenchyma. The IgA+ cells numbers increased time-dependently as compared with those in the control. The aggregation sizes and number of the F4/80+ cell in the splenic red pulp were increased. Furthermore the F4/80+ cells numbers increased time-dependently as compared with those in the control. The UEAI+ cells were found as free cells but not in aggregations in the control splenic red pulp. Contradictory to the number of IgA+ cells and F4/80+ cells the number of the UEAI+ cells decreased time-dependently after administration with SAS. Hematologically, abnormal numbers of WBCs different cells were observed after administration with SAS. This study provides new insight about the toxicity of a marine extract may be used in natural products industry or medical applications.