The Feasibility of Establishing a Morphological Cell Profiling Assay to Assess the Bioactivity of Natural Products

<p><b>Morphological cell profiling (MCP) is an assay which quantifies the morphology of cells and cellular components. Changes in the morphology of cells following compound treatment has been shown to assess the bioactivity, and even propose a target of said compound. It is a powerful assay that can be used to assess novel compounds for drug candidature. However, it is currently not transferable between institutions. Thus, this project evaluated the feasibility of establishing an MCP assay at Victoria University of Wellington - Te Herenga Waka to assess the bioactivity of novel natural products from the natural products laboratory.</b></p> <p>First, a new individualistic approach to select compound concentration for MCP was assessed by flow cytometry with six exemplary drugs. Results concluded this flow cytometry approach can realise an ideal concentration for individual compounds in which the compound treatment was bioactive but not cytotoxic. This approach provides more biologically relevant and accurate information about a compound's bioactivity than previous MCP methods.</p> <p>Second, the feasibility of establishing the assay was assessed by testing the same six drugs through protocol common to MCP assays. Various stains and microscopes were tested for suitability for an MCP; image quantitative software CellProfiler was evaluated for MCP analysis ease; and quality control protocol was attempted. Ultimately, an MCP is currently not feasible at Te Herenga Waka as multiple components to establish the assay are too difficult and time intensive to complete.</p> <p>Third, discorhabdin E, a pyrroloiminoquinone alkaloid, was isolated from the New Zealand marine sponge, Latrunculia kaakaariki. The compound was tested as if it were a novel natural product compound through the new flow cytometry concentration selection protocol to test its effectivity, and was subsequently assessed by morphological analysis.</p>

The Feasibility of Establishing a Morphological Cell Profiling Assay to Assess the Bioactivity of Natural Products

<p><b>Morphological cell profiling (MCP) is an assay which quantifies the morphology of cells and cellular components. Changes in the morphology of cells following compound treatment has been shown to assess the bioactivity, and even propose a target of said compound. It is a powerful assay that can be used to assess novel compounds for drug candidature. However, it is currently not transferable between institutions. Thus, this project evaluated the feasibility of establishing an MCP assay at Victoria University of Wellington - Te Herenga Waka to assess the bioactivity of novel natural products from the natural products laboratory.</b></p> <p>First, a new individualistic approach to select compound concentration for MCP was assessed by flow cytometry with six exemplary drugs. Results concluded this flow cytometry approach can realise an ideal concentration for individual compounds in which the compound treatment was bioactive but not cytotoxic. This approach provides more biologically relevant and accurate information about a compound's bioactivity than previous MCP methods.</p> <p>Second, the feasibility of establishing the assay was assessed by testing the same six drugs through protocol common to MCP assays. Various stains and microscopes were tested for suitability for an MCP; image quantitative software CellProfiler was evaluated for MCP analysis ease; and quality control protocol was attempted. Ultimately, an MCP is currently not feasible at Te Herenga Waka as multiple components to establish the assay are too difficult and time intensive to complete.</p> <p>Third, discorhabdin E, a pyrroloiminoquinone alkaloid, was isolated from the New Zealand marine sponge, Latrunculia kaakaariki. The compound was tested as if it were a novel natural product compound through the new flow cytometry concentration selection protocol to test its effectivity, and was subsequently assessed by morphological analysis.</p>

Advanced Optical Sensing of Phenolic Compounds for Environmental Applications

Phenolic compounds are particularly dangerous due to their ability to remain in the environment for a long period of time and their toxic effects. They enter in the environment in different ways, such as waste from paper manufacturing, agriculture (pesticides, insecticides, herbicides), pharmaceuticals, the petrochemical industry, and coal processing. Conventional methods for phenolic compounds detection present some disadvantages, such as cumbersome sample preparation, complex and time-consuming procedures, and need of expensive equipment. Therefore, there is a very large interest in developing sensors and new sensing schemes for fast and easy-to-use methods for detecting and monitoring the phenolic compound concentration in the environment, with special attention to water. Good analytical properties, reliability, and adaptability are required for the developed sensors. The present paper aims at revising the most generally used optical methods for designing and fabricating biosensors and sensors for phenolic compounds. Some selected examples of the most interesting applications of these techniques are also proposed.

Standardization of Mango (Mangifera Indica L.) Peel Simplisia of Gadung Variety

Mango peel (Mangfera indica L.) has many pharmacological effects as a traditional medicine. Therefore, standardization of mango peel simplisia needs to be done as a preparation of phytopharmaca raw material. This research aimed to obtain standardization of mango peel simplisia include specific and non-specific parameter. The research procedures include plant determination, simplisia preparation as well as specific standardization test (includes organoleptic, water-soluble compound concentration, and ethanol solution compound concentration) and nonspecific standardization test (includes moisture content, dried shrinkage, total ash content and acid insoluble ash content). The specific organoleptic parameters of dried mango peel simplisia have a distinctive sweet aroma, bitter taste, and brownish yellow colour. Water-soluble and ethanol-soluble concentrations are 22,36% ± 1,17% and 9,56% ± 0,07%. Moisture content is 9,09% ± 1,44%. Dried shrinkage rate is 0,19% ± 0,04%. Total ash and acid insoluble ash contents are 4,11% ± 0,10% and 0,14% ± 0,03%. The mango peel simplisia has met the quality standard of the raw material.

Evaluation of connectivity map shows limited reproducibility in drug repositioning

The Connectivity Map (CMap) is a popular resource designed for data-driven drug repositioning using a large transcriptomic compendium. However, evaluations of its performance are limited. We used two iterations of CMap (CMap 1 and 2) to assess their comparability and reliability. We queried CMap 2 with CMap 1-derived signatures, expecting CMap 2 would highly prioritize the queried compounds; the success rate was 17%. Analysis of previously published prioritizations yielded similar results. Low recall is caused by low differential expression (DE) reproducibility both between CMaps and within each CMap. DE strength was predictive of reproducibility, and is influenced by compound concentration and cell-line responsiveness. Reproducibility of CMap 2 sample expression levels was also lower than expected. We attempted to identify the “better” CMap by comparison with a third dataset, but they were mutually discordant. Our findings have implications for CMap usage and we suggest steps for investigators to limit false positives.

Cannabis Extract Has a Positive–Immunostimulating Effect through Proteolytic System and Metabolic Compounds of Honey Bee (Apis Mellifera) Workers

In the study, we assessed the effect of hemp extract on activities of resistance parameters and the metabolic compound concentration in adult workers’ hemolymph. Bees were divided into the following groups: (1) control group fed with mixture of sugar and water-glycerine solution, (2) experimental group with pure sugar syrup and inside with cotton strips soaked with hemp extract, (3) experimental group with a mixture of sugar syrup with hemp extract. Hemp extracts caused an increase in the protein concentrations and reduced the protease activities regardless of the administration method. The protease inhibitor activities were decreased only in the group that received hemp extract on the strips. The biomarker activities (ALP, ALT, AST) increased from the control group and workers feeding extract in syrup and decreased in workers supplemented with the extract on strips. In young, 2-day-old workers, the glucose concentration was higher in the groups feeding with the extract than in the control. Hemp extract influenced an increase in urea concentrations in workers’ hemolymph in comparison with the control. The hemp supplementation positively influences the immune system of workers, and the appropriate method of administration may be adapted to the health problems of bees.

Effect of salinity and pH on growth, phycoerythrin, and non-volatile umami taste active compound concentration of Rhodomonas salina using a D-optimal design approach

The cryptophyte Rhodomonas salina is widely used in aquaculture due to its high nutritional profile. This study aims to investigate the effect of salinity and pH on the growth, phycoerythrin concentrations, and concentrations of non-volatile umami taste active compounds of R. salina, using a design of experiment approach. Rhodomonas salina was cultivated in a flat-panel photobioreactor in turbidostat mode in a range of salinity (20–40 ‰) and pH (6.5–8.5). The strain was able to grow steadily under all conditions, but the optimal productivity of 1.17 g dry weight L−1 day−1 was observed in salinity 30 ‰ and pH 7.5. The phycoerythrin concentration was inversely related to productivity, presenting higher values in conditions that were not optimal for the growth of R. salina, 7% of dry weight at salinity 40 ‰, and pH 8.5. The identification of the umami taste of R. salina was based on the synergistic effect of umami compounds 5′-nucleotides (adenosine 5′-monophosphate, guanosine 5′-monophosphate, inosine 5′-monophosphate) and free amino acids (glutamic and aspartic acids), using the equivalent umami concentration (EUC). The results indicated that an increase in pH induces the accumulation of 5′-nucleotides, resulting in an EUC of 234 mg MSG g−1 at a salinity of 40 and pH 8.5. The EUC values that were observed in R. salina were higher compared to other aquatic animals, a fact that makes R. salina promising for further research and application in the food and feed sectors.

Effect of blue, green or red LED light on the functional quality of spinach (Spinacia oleracea L.)

The present study employed white (W), blue (B: 468 nm), red (R: 629 nm) and green (G: 524 nm) monochromatic LED lights for 26 days, from 11:00 to 18:00 (7 h per day), with a average of photosynthetic photon flux density (PPFD) of 26.00 m-2 s-1 on two baby leaves of spinach (Spinacia oleracea L.) cultivars (Falcon F1 and Viroflay) grown in a hydroponic system. Regardless of the cultivar, the fresh and dry weights were positively influenced when the plants were irradiated by R-light in comparison to W-light. Independent of the cultivar, the leaves treated with B-light reached a significantly higher phenolic compound concentration than plants irradiated with W-light. With respect to antioxidant capacity, plants treated with B-light showed a significantly higher antioxidant capacity than those treated with W-light. According to the results, the use of LED lights is a promising technique for the production of antioxidant compound-enriched vegetables. Highlights - The fresh and dry weights were positively influenced when baby leaves spinach were irradiated by red LED light. - Baby leaves spinach treated with blue LED light reached a significantly higher phenolic compound concentration and antioxidant capacity. - The green LED light increased total phenolic compound concentration in baby leaves spinach.

Domestication of Plants of Ugni molinae Turcz (Myrtaceae) Interferes in the Biology of Chilesia rudis (Lepidoptera: Erebidae) Larvae

In terms of the domestication process in murtilla, studies have found changes in the concentration of phenolic compounds, with reduction of chemical defense of plants, depending on the change in the feeding behavior of insects. Thus, we hypothesized that the domestication of Ugni molinae decreases the content of phenolic compounds and modifies the feeding preference of Chilesia rudis larvae. Leaves of three parental ecotypes and four cultivated ecotypes were used in preference experiments to evaluate the mass gain and leaves consumption of larvae. Phenolic extracts from leaves of U. molinae were analyzed by HPLC. Identified compounds were incorporated in an artificial diet to assess their effect on mass gain, consumption, and survival of the larvae. The presence of phenolic compounds in bodies and feces was also evaluated. In terms of choice assays, larvae preferred parental ecotypes. Regarding compounds, vanillin was the most varied between the ecotypes in leaves. However, plant domestication did not show a reduction in phenolic compound concentration of the ecotypes studied. Furthermore, there was no clear relation between phenolic compounds and the performance of C. rudis larvae. Whether this was because of sequestration of some compounds by larvae is unknown. Finally, results of this study could also suggest that studied phenolic compounds have no role in the C. rudis larvae resistance in this stage of murtilla domestication process.