Visual Replacement Projects

Contains reports on two research projects

Determining the Minimum Inhibitory Concentration of Fulvic Acid on Early Streptococcus mutans Biofilm Formation

poster abstractFulvic acid is an active ingredient in many homeopathic treatments. Shilajit is one such treatment that is mainly used in the Indian Subcontinent area. It is believed that fulvic acid has some remedial properties that can help with inhibition of many types of bacteria and various malignant diseases. In this experiment the effects of fulvic acid were analyzed on early Streptococcus mutans biofilm formation. S. mutans is an oral bacterium that contributes to the formation of dental caries forming bacterial biofilm on teeth. For the experiment, early S. mutans biofilm formation was treated with different concentrations of fulvic acid for 24 hours in sterile 96-well flat-bottom microtiter plates. The optical density (OD) of the S. mutans biofilm was then measured using a SpectraMax190, after staining with crystal violet. Data was analyzed on the knowledge that a greater OD is correlated to a greater bacterial biofilm. Results demonstrated that fulvic acid inhibited the growth of early S. mutans biofilm formation at fulvic acid concentrations greater than 5% (vol. %). After preliminary studies, different concentrations of fulvic acid closer to the estimated minimum inhibitory concentration (MIC) were applied to the S. mutans to find a more precise MIC of the fulvic acid on the biofilm growth. Upon completion of these various studies, fulvic acid was shown to inhibit early S. mutans biofilm formation and may show signs of oral health improvement if applied for human use

Evaluating the Effect of Fulvic Acid on Oral Bacteria and Cancerous Oral Cells

poster abstractShilajit is a homeopathic treatment used by local inhabitants of India and Pakistan. It may have specific components that inhibit the formation of cavities and the growth of cancer cells. This experiment analyzed the effects of fulvic acid, an active component of shilajit, on the growth of oral bacteria and squamous cell carcinoma. The effect of fulvic acid was evaluated on early Streptococcus mutans (S. mutans) biofilm formation and established S. mutans biofilm by treating each group with different concentrations of fulvic acid for 24 hours in sterile 96-well flat-bottom microtiter plates. S. mutans was used because it is a common cause of dental caries. The optical density (OD) of the S. mutans biofilm was measured after crystal violet staining using a SpectraMax190; greater growth correlated to greater OD. It was determined that fulvic acid inhibits the growth of newly forming S. mutans biofilm at fulvic acid concentrations greater than 1.25% (vol. %) and established S. mutans biofilm at fulvic acid concentrations greater than 5% (vol. %). To evaluate the effect of fulvic acid on squamous cell carcinoma (SCC-25) cells, six-well plates seeded with SCC-25 cells (1*105 cells/well) were exposed to different concentrations of fulvic acid (buffered to a pH of 7.5) for 72 hours. The cytotoxicity and cell proliferation were measured using a cytotoxicity detection kit and a water soluble tetrazolium kit (Roche Applied Science), respectively. It was determined that fulvic acid inhibits the growth of SCC-25 cells at concentrations of fulvic acid above 2% (volume %). The effects of fulvic acid (0.5%) on matrix metalloproteinase expression and collagen degradation ability of SCC-25 cells is being analyzed. The suppressive mechanisms observed by fulvic acid on both S. mutans and SCC-25 cells could improve overall oral health

Sensory Replacement

Contains a report on a research project.Purchase Order DDL-B15

Visual Replacement Projects

Contains reports on two research projects

The Addition of Arachidin 1 or Arachidin 3 to Human Rotavirus-infected Cells Inhibits Viral Replication and Alters the Apoptotic Cell Death Pathway

Rotavirus (RV) infections are a leading cause of severe gastroenteritis in infants and children under the age of five. There are two vaccines available in the United States and one in India that can be administered early in childhood, however they only protect against specific strains1. From our previous work, both arachidin-1 (A1) and arachidin-3 (A3) from peanut (Arachis hypogaea) hairy root cultures significantly inhibit simian RV replication2,3,4. The purpose of this study was to determine if a human intestinal cell line, HT29.f8, infected with a human RV, Wa, was affected by A1 and A3. Cell viability assays were utilized to determine if A1 and A3 affect the HT29.f8 cells with/without RV infections. At eighteen hours post infection (hpi), supernatants from the RV-infected HT29.f8 cells with/without the arachidins were used in plaque forming assays to quantify and compare the amount of infectious RV particles that are produced during an infection. Transmission electron microscopy (TEM) was used to visualize cell ultrastructure and individual RV particles. Additionally, tunable resistive pulse sensing technology (TRPS) using the qNano system by IZON was employed to quantify and measure virus particle sizes, and display the size distribution of RV particles. Likewise, quantitative real time polymerase chain reactions (qRT-PCR) were performed to determine if A1 and A3 regulated cell death pathways in the HT29.f8 cell line. This data will guide our future studies to determine the antiviral mechanism(s) of action of A1 and A3

Improving bone properties and fracture susceptibility: experimental models of genetic manipulation, pharmacologic intervention, and cellular perturbation reveal new approaches for improving bone health

poster abstractBone, a crucial support structure in the human body, is often taken for granted for its lightweight properties and unparalleled strength. Skeletal fracture is a major clinical condition affecting millions of Americans, which results from abnormal aging, hormonal imbalance, genetic conditions, and lifestyle choices (e.g., exercise). Because fractures are caused by a number of different factors, reducing fracture incidence requires a multifactorial approach to unraveling the underlying biology of bone metabolism, in order to discover new ways to improve bone properties and prevent fractures. We have taken such an approach by conducting (1) genetic manipulation experiments in mice, where genes predicted to be involved in bone mass regulation were mutated; (2) pharmacologic experiments to quantify the dose-response effect of an agent that inhibits bone loss, and (3) cell culture experiments, aimed at revealing molecular pathways activated by mechanical stimulation. METHODS: Mice with mutations in two genes, likely to regulate bone mass (SOST, DKK1) were generated and subjected to in vivo dual energy x-ray absorptiometry (DEXA) scans at 6-wk old. Whole body scans were analyzed for bone mineral density (BMD) using Lunar Piximus II v2.10 software. Mice (6-wk) were also dosed (0, 1, 10, 100, or 1000 mg/kg) with daily alendronate HCl, a bisphosphonate that inhibits osteoclast activity. Six wks later, the mice were sacrificed, and the femurs were dissected and sectioned for histological analysis of bone formation parameters, including mineralizing surface (MS/BS), mineral apposition rate (MAR), and bone formation rate (BFR/BS). To understand the cellular signaling events in response to mechanical loading, bone marrow mesenchymal stem cells (MSCs) were treated with 10, 20, 30, or 40μM PF7408671, an S6 kinase inhibitor. Cells then were subject to 100 cycles of biaxial mechanical strain (2%, 10 cycles/min). Protein lysates were separated by electrophoresis and probed for phosphorylation of Rictor and Akt by Western blot. RESULTS: Mice harboring mutations in either the SOST gene or the DKK1gene exhibited significantly increased BMD compared to wild-type control mice, though the SOST mutation had a stronger effect on BMD than DKK1. Mice with compound mutations (SOST and DKK1 mutations) had significantly greater BMD than mice with either single mutation, suggesting that inhibition of SOST and DKK1 might be an effective means to increase bone mass in patients susceptible to fracture. Mice treated with high-dose alendronate (100 or 1,000 mg/kg) exhibited significant decreases in bone formation parameters (MS/BS, MAR, and BFR/BS) compared to untreated (0 mg) mice, suggesting that while this compound might be beneficial for inhibiting bone loss, it also inhibits bone formation. The signaling hub, mTORC2, is a critical regulator of mechanical force in MSC progenitors. Our data demonstrate that S6 kinase is an upstream activator of mTORC2 in response to mechanical strain. CONCLUSION: Our experiments suggest that genetic manipulation of mice reveal viable protein targets (e.g., SOST, DKK1) that could ultimately be manipulated pharmacologically to improve bone mass. We also found that an FDA-approved class of drugs inhibits bone formation even at very low doses, suggesting that additional pro-anabolic compounds might benefit patients taking bisphosphonates. On a cell signaling level, we found that the mTORC2 pathway shows considerable promise for pharmacologic manipulation to simulate the effects of exercise. Taken together, these experiments highlight the utility of a broad approach to solving bone metabolism challenges that can affect fracture susceptibility

Asset Allocation with Swarm/Human Blended Intelligence

PSO has been used to demonstrate the near-real-time optimization of frequency allocations and spatial positions for receiver assets in highly complex Electronic Warfare (EW) environments. The PSO algorithm computes optimal or near-optimal solutions so rapidly that multiple assets can be exploited in real-time and re-optimized on the fly as the situation changes. The allocation of assets in 3D space requires a blend of human intelligence and computational optimization. This paper advances the research on the tough problem of how humans interface to the swarm for directing the solution. The human intelligence places new pheromone-inspired spheres of influence to direct the final solution. The swarm can then react to the new input from the human intelligence. Our results indicate that this method can maintain the speed goal of less than 1 second, even with multiple spheres of pheromone influence in the solution space