The Effect of Luteolin on Human Glioblastoma

Glioblastoma multiforme (GBM) is widely recognized as the most common and lethal of the malignant gliomas. Few effective therapeutic treatments are available as five-year survival rates of diagnosed individuals are less than five percent. Luteolin, a common flavonoid found in a variety of fruits and vegetables, has demonstrated significant promise in combating cancers of the breast, colon, liver, lung, and bone. In this study, we investigated the effects of luteolin on glioblastoma multiforme cell lines U-251, U-87, and U-1242. Cell viability was assessed using cell count with trypan blue exclusion and MTT assays. Results revealed that luteolin reduces GBM cell viability and cell proliferation in a time and concentration-dependent manner. Western Blot analysis indicated that luteolin decreased AKT, ERK, and MAPK phosphorylation following treatment with EGF. Additionally, luteolin promoted apoptosis in GBM cells by inducing PARP and caspase-3 cleavage, and decreasing levels of the anti-apoptotic protein BCL-XL. Our results indicate that luteolin exhibits a biological effect and may be used as a therapeutic agent for glioblastoma multiforme

Investigation of the Effects of Growth Environment on the Ferric Reducing Antioxidant Power of Selected Plant Species

Metabolism within the human body creates multiple oxidant by-products. These oxidants may cause cell injury, damage to DNA, and other complications leading to the development of chronic disease. Antioxidants are important dietary components which defend against oxidative damage by scavenging the oxidant by-products. Research has shown that diets rich in antioxidants offer protection against various chronic diseases. The goal of this research is to determine the effects of varying growing conditions on the production of antioxidants, and to ultimately find the best possible plant-growth environment for maximum production of antioxidants. Each plant was grown under three different environmental conditions; positive, negative, and control treatment. The positive treatment consisted of supplying water to field capacity with fertilizer, the negative treatment consisted supplying half of the water required to reach field capacity with no fertilizer, and the control treatment consisted of supplying water to field capacity with no fertilizer. Ferric reducing antioxidant levels were then determined. The ferric reducing antioxidant power evaluates antioxidant potential by reducing ferric iron (Fe3+) to its ferrous form (Fe2+). Addition of excess ferric ions result in the development of a Prussian blue color. The ferric reducing antioxidant power of the extracts was measured by reading the absorbance at 750 nm using a spectrophotometer. The ferric reducing antioxidant power assay was performed on extracts of red clover (Trifolium pratense), Amur honeysuckle (Lonicera maackii) and wild garlic (Allium vineale). The differing growing conditions resulted in variation in the production of antioxidants by the plants. The data obtained revealed that the plants grown under the negative treatment produced a significantly lower level of antioxidants when compared to the plants grown under the positive treatment. These results indicate that growing conditions can influence antioxidant production in plants

Antinociceptive Effects of Herkinorin, a MOP Receptor Agonist Derived from Salvinorin A in the Formalin Test in Rats: New Concepts in Mu Opioid Receptor Pharmacology

Herkinorin is the first μ opioid (MOP) selective agonist derived from salvinorin A, a hallucinogenic natural product. Previous work has shown that, unlike other opioids, herkinorin does not promote the recruitment of β-arrestin-2 to the MOP receptor and does not lead to receptor internalization. This paper presents the first in vivo evaluation of herkinorin’s antinociceptive effects in rats, using the formalin test as a model of tonic inflammatory pain. Herkinorin was found to produce a dose-dependent decrease in the number of flinches evoked by formalin. These antinociceptive effects were substantially blocked by pretreatment with the nonselective antagonist naloxone, indicating that the antinociception is mediated by opioid receptors. Contralateral administration of herkinorin did not attenuate the number of flinches evoked by formalin, indicating that its effects are peripherally restricted to the site of injection. Following chronic administration (5-day), herkinorin maintained antinociceptive efficacy in both phases of the formalin test. Furthermore, unlike morphine, herkinorin was still able to inhibit flinching in both phases of the formalin test in animals made tolerant to chronic systemic morphine treatment. Collectively, these results suggest that herkinorin may produce peripheral antinociception with decreased tolerance liability and thereby represents a promising template for the development of agents for the treatment of a variety of pain states

The Effects of Apigenin on Cell Proliferation and Apoptosis in Glioblastoma Multiforme

Glioblastoma multiforme (GBM) is a WHO grade IV brain tumor. These tumors are highly proliferative, infiltrative, necrotic, angiogenic, and resistant to apoptosis. One major characteristic of GBM is the overexpression of epidermal growth factor receptor (EGFR), which leads to cell growth and proliferation when activated. GBM is very difficult to treat due to its location, heterogeneity, and invasiveness; an effective treatment is therefore needed. The use of flavonoids, which are natural compounds found in many fruits and vegetables, has been studied in the treatment of many different tumor types. Apigenin is a specific flavonoid that has previously been shown to have antitumor activity in a number of cancer cells. Our study set out to investigate the molecular effects of apigenin treatment on glioblastoma cell proliferation and viability using the trypan blue exclusion assay, MTT assay, and an LDH assay. In addition, Western blot analyses were utilized out to determine the signaling pathways through which apigenin treatment exerts its effects on cell proliferation and apoptosis. Finally, hoechst-propidium iodide staining and flow cytometry were used to examine the extent of apoptosis and the cell cycle context of these effects. Our results show that apigenin reduces cell viability and proliferation in a dose and time dependent manner while increasing cytotoxicity in GBM cells. Additionally, apigenin inhibits the EGFR mediated phosphorylation in the presence of EGF treatment of AKT, mTOR, and s6k resulting in decreased cell survival, growth and proliferation. It also inhibits the MAPK pathways in one cell line thereby reducing cell growth and proliferation. It also inhibits the anti-apoptotic effects of BCL-XL and increases PARP cleavage, which leads to increased apoptosis. Finally, apigenin induced cycle arrest at the G2M checkpoint, meaning that apoptosis primarily occurred at the DNA repair checkpoint in the cell cycle. In conclusion, apigenin has demonstrated some in vitro biological effects on glioblastoma cell lines that show promises in limiting the growth, proliferation and survival of these cell lines. Future research should look to identify means through which apigenin can be administered in clinically significant concentrations to the brain

Natural Products as Therapeutic Agents in Cancer Treatment

Cancer accounts for 25% of deaths in the United States, and brain tumors greatly contribute to this percentage. However, relative to other types of cancers, brain tumors prove difficult to treat because they are heterogeneous, highly proliferative, highly invasive, and resistant to the traditional cancer treatments of chemotherapy and radiotherapy. Past studies have shown that flavonoids and curcuminoids, two classes of compounds derived from natural sources, are effective in inhibiting the development and metastasis of breast and lung cancer cells. Research has also indicated that these compounds have potential for treating brain tumors. The purpose of this research is to further explore the potential of flavonoids as therapeutic options for the treatment of brain tumors. Specifically, flavonoids’ effect on cell proliferation, cell death, and tumor invasion will be studied. Another objective of this study is to identify the signaling mechanism by which flavonoids mediate their therapeutic effects on brain tumor cell lines. Three human brain tumor cell lines (U-1242, U-251, and U-87) will be studied. They will be treated with various flavonoids at increasing concentrations (10, 20, 40, and 80 µM). Cells will be counted following the trypan blue staining protocol. MTT assays and Western blot analyses will be used to assess cell proliferation. Cell death will be assessed with flow analyses and Western blot analyses. Unpaired t-tests will be run to compare treated and control cells at a 95% confidence interval. If necessary, one-way ANOVA with multiple comparisons will be used to compare multiple treatment groups and a control at a 95% confidence interval, and the Tukey post-hoc test will be utilized if appropriate. All statistical tests will be run in IBM SPSS 21®

Correlation Study: Student Success in Biochemistry as a Prerequisite for Integrated Pharmacology and Medicinal Chemistry

Background A rise in new schools of pharmacy has led to implementation of new curriculums. Pharmacy schools must adhere to standards set by the Accreditation Council of Pharmacy Education in order to provide knowledge of foundational sciences and prepare pharmacy students for the future. Prerequisites are typically foundational science courses taken early in the program so that students have the knowledge necessary to be excellent pharmacists. Within the Cedarville University School of Pharmacy, Biochemistry is a prerequisite course for Integrated Medicinal Chemistry and Pharmacology (PCoMedChem). Objectives The goal of this study is to determine if Biochemistry should remain a prerequisite course for Integrated Medicinal Chemistry and Pharmacology at Cedarville University under a TBL setting based on if student success in Biochemistry influences student success in PcoMedChem. Methodology The study will evaluate student individual and overall course grades for both Biochemistry and PcoMedChem. The data will include grades from the 2018-2021 cohorts of pharmacy students. Inclusion criteria consists of completion of Biochemistry and Integrated Medicinal Chemistry and Pharmacology. No exclusion of students exist because all student data will be evaluated. Students will complete a survey through Qualtrics regarding extracurricular commitments as well as perceptions towards the courses to supplement the findings and explain discrepancies. Analysis The demographics and students’ perceptions will be compared in SPSS by analyzing frequency of responses. Using SPSS, the Wilcoxon test and Levene’s test will be conducted followed by a Pearson or Spearman correlation, depending on distribution, in order to determine correlation between grades in Biochemistry and PcoMedChem. Additionally, an ANCOVA test will be used to analyze the data gathered from our survey. A p-value of 0.05 will be indicative of statistical significance

The Effects of Herkinorin, the First mu-Selective Ligand from a Salvinorin A-Derived Scaffold, in a Neuroendocrine Biomarker Assay in Nonhuman Primates

Herkinorin is the first μ-opioid receptor-selective ligand from the salvinorin A diterpenoid scaffold. Herkinorin has relative μ \u3e κ \u3e δ binding selectivity, and it can act as an agonist at both μ- and κ-receptors, in vitro. These studies were the first in vivo evaluation of the effects of herkinorin in nonhuman primates, using prolactin release, a neuroendocrine biomarker assay that is responsive to both μ- and κ-agonists, as well as to compounds with limited ability to cross the blood-brain barrier. In cumulative dosing studies (0.01–0.32 mg/kg i.v.), herkinorin produced only small effects in gonadally intact males (n = 4), but a more robust effect in females (n = 4). Time course studies with herkinorin (0.32 mg/kg) confirmed this greater effectiveness in females and revealed a fast onset after i.v. administration (e.g., by 5–15 min). Antagonism experiments with different doses of nalmefene (0.01 and 0.1 mg/kg) caused dose-dependent and complete prevention of the effect of herkinorin in females. This is consistent with a principal μ-agonist effect of herkinorin, with likely partial contribution by κ-agonist effects. The peripherally selective antagonist quaternary naltrexone (1 mg/kg s.c.) caused approximately 70% reduction in the peak effect of herkinorin (0.32 mg/kg) in females, indicating that this effect of herkinorin is prominently mediated outside the blood-brain barrier

Evaluating Antioxidant Activity of Selected Plant Species Native to Cedarville, Ohio

Over the past several decades, there has been an increase in the number of synthetic drug molecules developed and utilized to treat various conditions. Although these synthetic drugs have proven useful, there has been growing public concern regarding the potentially negative long-term effects of synthetic agents on the body. As a result, there has been an increased interest in identifying and utilizing plant extracts and purified compounds since they are perceived to be a more natural alternative to synthetic drugs. The goal of this study was to evaluate the specific antioxidant properties of alsike clover Trifolum hybridum when produced under differing growing conditions. The alsike clover was collected from the campus of Cedarville University, Cedarville, Ohio for testing. Alsike clover was removed from the field in January 2013, and transplanted indoors under grow lights for 14 days. These plants were then subjected to three separate 60-day treatments: control treatment - watering to field capacity with no fertilizer; positive treatment - watering to field capacity with fertilizer; and negative treatment - half of the water given to the field capacity treatment with no fertilizer. The rationale for choosing these different treatments was to evaluate the effects of specific growing conditions on bioactive secondary metabolite production in alsike clover. The biological evaluation was accomplished by conducting diphenylpicrylhyrazyl (DPPH) free-radical scavenging and Folin Ciocalteu assays to assess the concentration of polyphenolic compounds. Results from these experiments indicate that the biological and chemical profiles of alsike clover can be influenced by the environmental conditions under which the plants are grown

Natural Products as Therapeutic Agents in Cancer Treatment

Cancer accounts for 25% of deaths in the United States, and brain tumors greatly contribute to this percentage. However, relative to other types of cancers, brain tumors prove difficult to treat because they are heterogeneous, highly proliferative, highly invasive, and resistant to the traditional cancer treatments of chemotherapy and radiotherapy. Past studies have shown that flavonoids and curcuminoids, two classes of compounds derived from natural sources, are effective in inhibiting the development and metastasis of breast and lung cancer cells. Research has also indicated that these compounds have potential for treating brain tumors. The purpose of this research is to further explore the potential of flavonoids as therapeutic options for the treatment of brain tumors. Specifically, flavonoids’ effect on cell proliferation, cell death, and tumor invasion will be studied. Another objective of this study is to identify the signaling mechanism by which flavonoids mediate their therapeutic effects on brain tumor cell lines. Three human brain tumor cell lines (U-1242, U-251, and U-87) will be studied. They will be treated with various flavonoids at increasing concentrations (10, 20, 40, and 80 µM). Cells will be counted following the trypan blue staining protocol. MTT assays and Western blot analyses will be used to assess cell proliferation. Cell death will be assessed with flow analyses and Western blot analyses. Unpaired t-tests will be run to compare treated and control cells at a 95% confidence interval. If necessary, one-way ANOVA with multiple comparisons will be used to compare multiple treatment groups and a control at a 95% confidence interval, and the Tukey post-hoc test will be utilized if appropriate. All statistical tests will be run in IBM SPSS 21®