Effects of Cannabidiol on MCF-7 Human Breast Cancer Cell Proliferation, Metabolism, and Morphology

The use of cannabis for both recreational and medicinal purposes is not new or novel. Of the diverse class of compounds found in cannabis, cannabidiol (CBD) has been shown in previous studies to impede cancer cell growth. The ability of CBD to inhibit cancer cell proliferation has wide implications for both cannabis and cancer. The purpose of this study was to determine CBD’s ability to affect MCF-7 human breast cancer cell proliferation and morphology. Briefly, pure CBD was extracted using hexane in two simple chromatographic columns and used to treat subconfluent MCF-7 human breast cancer and C2C12 mouse myoblast cell cultures. MCF-7 and C2C12 cell lines were treated with standard solutions of 1, 2, 10, 50, 100, and 200 ug/mL CBD. MCF-7 and C2C12 metabolism and proliferation was quantitatively analyzed using MTT and hemacytometer counts, respectively and compared across all CBD concentrations. MCF-7 and C2C12 cell morphology was qualitatively evaluated using fluorescence and inverted light microscopy. Results showed that CBD-treated MCF-7 cancer cells experienced decreased proliferation, reduced size, and changed morphology, whereas C2C12 myoblast cells showed negligible decreases in proliferation and morphological changes. The implications of this study show a possible alternative use of CBD for treatment of cancer with reduced non-cancerous cell toxicity

Application of \u3ci\u3eDalea ornata\u3c/i\u3e (Fabaceae) extractives toward inhibition of the hookworm \u3ci\u3eAncylostoma ceylanicum\u3c/i\u3e

Under the principal investigator Dr. Gil Belofsky of the Department of Chemistry, this research project is focused on the discovery and application of novel compounds from the plant Dalea ornata, commonly known as the Blue Mountain prairie clover. The process of isolating each compound was accomplished through techniques such as Sephadex LH-20 size exclusion chromatography and linear continuous gradient chromatography over silica gel. Eight different phenolic compounds have been isolated, with several more nearly pure. Advanced nuclear magnetic resonance spectroscopy techniques were utilized to elucidate the structure of each isolated compound. In the event that additional amounts are needed, re-isolation of compounds will be performed for expanded biological testing. Initial ex vivo biological testing of D. ornata extracts have demonstrated anthelmintic activity towards the hookworm Ancylostoma ceylanicum. The World Health Organization projects that one billion individuals are currently infected with hookworms or related soil-transmitted, parasitic nematodes. While medications are available to treat acute cases, there is no current treatment to prevent hookworm infection. Determination of the D. ornata compound(s) associated with the observed anthelmintic activity will likely provide further insight into mechanisms of inhibiting and preventing hookworm infections

Chromatographic Isolation and Characterization of Secondary Metabolites of Dalea ornata (Fabaceae)

The process of isolating and characterizing natural products is a vital component of drug discovery, through which the pharmaceutical industry drives the creation of synthetic blockbuster drugs like Lipitor© and Paxil©. Advanced methods in column chromatography including polarity-based gradient elution, neutral size-exclusion polymers, and effective techniques in visualization using thin layer chromatography contribute to the successful purification of novel natural products. In our work to date, a number of phenolic compounds have been isolated from the shrub Dalea ornata (Fabaceae), and at cellular-level concentrations show efficacy eradicating the problematic parasite Ancylostema ceylanicum, or hookworm. Our research is toward the continued isolation of relatively abundant and chemically-viable target molecules, by various chromatographic methods. This approach frequently yields newly discovered compounds of many varieties, possibly including antibiotic tetracyclines and colorful flavonoids. Nuclear magnetic resonance spectroscopy, mass spectrometry, polarimetry, and other physicochemical methods of analysis are utilized to determine the compounds’ chemical characteristics in the Department of Chemistry, in conjunction with anthelmintic or anti-hookworm ex vivo testing, carried out in collaboration with Dr. Blaise Dondji’s group in the Department of Biological Sciences

Regional assessment on influence of landscape configuration and connectivity on range size of white-tailed deer

Variation in the size of home range of white-tailed deer (Odocoileus virginianus) has broad implications for managing populations, agricultural damage, and disease spread and transmission. Size of home range of deer also varies seasonally because plant phenology dictates the vegetation types that are used as foraging or resting sites. Knowledge of the landscape configuration and connectivity that contributes to variation in size of home range of deer for the region is needed to fully understand differences and similarities of deer ecology throughout the Midwest. We developed a research team from four Midwestern states to investigate how size of home range of deer in agro-forested landscapes is influenced by variations in landscape characteristics that provide essential habitat components. We found that for resident female deer, annual size of home range in Illinois (mean = 0.99 km2), Michigan (mean = 1.34 km2), Nebraska (mean = 1.20 km2), and Wisconsin (mean = 1.47 km2) did not differ across the region (F3,175 = 0.42, P = 0.737), but differences between agricultural growing and non-growing periods were apparent. Variables influencing size of home range included: distance to forests, roads, and urban development from the centroid of deer home range, and percent of crop as well as four landscape pattern indices (contrast-weighted edge density, mean nearest neighbor, area-weighted mean shape index, and patch size coefficient of variation). We also identified differences in model selection for four landscapes created hierarchically to reflect levels of landscape connectivity determined from perceived ability of deer to traverse the landscape. Connectivity of selected forested regions within agro-forested ecosystems across the Midwest plays a greater role in understanding the size of home ranges than traditional definitions of deer habitat conditions and landscape configuration

Phenolic Metabolites of Dalea ornata Affect Both Survival and Motility of the Human Pathogenic Hookworm Ancylostoma ceylanicum

Hookworms are ubiquitous human parasites, infecting nearly one billion people worldwide, and are the leading cause of anemia and malnutrition in resource-limited countries. Current drug treatments rely on the benzimidazole derivatives albendazole and mebendazole, but there is emerging resistance to these drugs. As part of a larger screening effort, using a hamster-based ex vivo assay, anthelmintic activity toward Ancylostoma ceylanicum was observed in the crude extract of aerial parts of Dalea ornata. These studies have led to the isolation and characterization of phenolic metabolites 1–10. The structures were determined by 1D and 2D NMR spectroscopy, and the absolute configuration of 1 was assigned using electronic circular dichroism data. The new compound, (2S)-8-(3-methylbut-2-en-1-yl)-6,7,4′-trihydroxyflavanone, was weakly active at 7.3 μM, with 17% reduction in survival of the hookworms after 5 days. The rotenoids deguelin and tephrosin, predictably perhaps, were the most active, with complete worm mortality observed by day 4 (or earlier) at 6.3 and 6.0 μM, respectively. The effects of 1–10 on hookworm motility and on toxicity to hamster splenocytes were also explored as important measures of treatment potential

Phenolic Metabolites of <i>Dalea ornata</i> Affect Both Survival and Motility of the Human Pathogenic Hookworm <i>Ancylostoma ceylanicum</i>

Hookworms are ubiquitous human parasites, infecting nearly one billion people worldwide, and are the leading cause of anemia and malnutrition in resource-limited countries. Current drug treatments rely on the benzimidazole derivatives albendazole and mebendazole, but there is emerging resistance to these drugs. As part of a larger screening effort, using a hamster-based ex vivo assay, anthelmintic activity toward <i>Ancylostoma ceylanicum</i> was observed in the crude extract of aerial parts of <i>Dalea ornata</i>. These studies have led to the isolation and characterization of phenolic metabolites <b>1</b>–<b>10</b>. The structures were determined by 1D and 2D NMR spectroscopy, and the absolute configuration of <b>1</b> was assigned using electronic circular dichroism data. The new compound, (2<i>S</i>)-8-(3-methylbut-2-en-1-yl)-6,7,4′-trihydroxyflavanone (<b>1</b>), was weakly active at 7.3 μM, with 17% reduction in survival of the hookworms after 5 days. The rotenoids deguelin (<b>9</b>) and tephrosin (<b>10</b>), predictably perhaps, were the most active, with complete worm mortality observed by day 4 (or earlier) at 6.3 and 6.0 μM, respectively. The effects of <b>1</b>–<b>10</b> on hookworm motility and on toxicity to hamster splenocytes were also explored as important measures of treatment potential