George Washington Brownridge III, Sylvanna Islas, Angelina Miller, and Warren Hoeffler
Bone constantly cycles through a dynamic process of breakdown and remodeling. Osteoblasts are the specialized mesenchymal stem cells that have a major role in bone formation and the remodeling process whereas their counterpart osteoclasts, handle bone resorption. Embryonic stem cells can be partially differentiated into Progenitor cells, and we worked with #18, a candidate for being an osteoprogenitor that has the potential to respond to morphogenic activators. In the case of bone remodeling, TGF-β 2, BMP-2 and an abundance of CA++ have been shown to be potential activators of differentiation into osteoblasts. Eight different trials were conducted with the cells using different combinations of the three morphogenic activators. After inducing the cells with the activators, we performed Immunohistochemistry (IHC) to analyze the expression of osteocalcin, which is the enzyme that binds calcium to mineralize bone. The cells with varying activator combinations showed different physiology with a variance in the cell shape, structure, and spacing. The greatest results were from the combination of TGF-β 2 and BMP-2, which is consistent with #18 operating as an osteoprogenitor. A 3D construct model of #18 seemed to have a similar structure to that of an osteon, possibly indicating the formation of bone. We took slices of the model and performed an IHC staining for Osteocalcin, Prolyl Hydroxylase (5B5), and Collagen I. We saw a strong positive signal for Col I and 5B5, and a slight positive signal for Osteocalcin. This information confirmed that #18 is an osteoprogenitor and is able to assemble bone.
Scale Up Isolation of Aaptamine for In Vivo Evaluation Indicates Its Neurobiological Activity is Linked to the Delta Opioid Receptor
Nicole L. McIntosh, Eptisam Lambo, Laura Millan-Lobo, Fei Li, Li He, Phillip Crews, Jennifer L. Whistler, and Tyler Johnson
Opioid receptors belong to the large superfamily of seven transmembrane-spanning (7TM) G protein-coupled receptors (GPCRs). As a class, GPCRs are of fundamental physiological importance mediating the actions of the majority of known neurotransmitters and hormones. The Mu, Delta, and Kappa (MOP, DOP, KOP) opioid receptors are particularly intriguing members of this receptor family as they are the targets involved in many neurobiological diseases such as addiction, pain, stress, anxiety, and depression. Recently we discovered that the aaptamine class of marine sponge derived natural products exhibit selective agonist activity in vitro for the DOP versus MOP receptor. Our findings may explain reports by others that aaptamine demonstrates in vivo anti-depressant effects in mouse models using the Porsolt Forced Swim Test. This project involved the extraction of the sponge Aaptos aaptos (a source of 1), establishing a scale up purification procedure to provide sufficient amounts of 1 (30 mg) for a follow up in vivo evaluation and ultimately confirmation of the structure of 1 using LC-MS and 1H NMR. The results our purification scheme, chemical analysis and in vivo evaluation of 1 using the Marble burying test in rodents are reported here in and suggest that the in vivo anti-depressant effects of 1 are linked directly to its agonist effects on the DOP receptor.
Alexandra Ham, Gabrielle Pecora, Hoaithuong Bui, Timothy Camarella, Victor Pham, and Marc Ting
The global energy economy is huge and thoughts of replacing large amounts of petroleum based fuels by massive levels of fermentation of grains are not realistic. On an energy basis what global agriculture produces for food will almost cover the energy demands if all of it is redirected to the production of fuels—either as alcohols for gasoline or as fat derivatives for diesel fuel. This means that chemical processes need to be developed that allow inclusion of non-food based agricultural and urban wastes as well as forest debris into the energy economy. These represent opportunities to capture new sources of energy that would otherwise not be captured. This project is based on the idea that every little bit helps, and focuses on a hands-on approach to isolating chemicals from fallen vegetation with an emphasis on adding to the transportation fuel pool. Hydrolysis of cellulosic wastes from various sources easily collected on our campus has been explored seeking ways to break them down to fermentable sugars. These sugars are then fermented to form alcohols suitable for inclusion in gasoline. Extraction of vegetable oils has also been explored. Finally an attempt has been made to quantify the impact such a strategy might have on global energy supplies if practiced on a wide-scale basis.
Sage Callaway-Keeley and Stephanie Huynh
The most effective ways to reduce CO2 emissions are to improve the energy efficiency of each economic sector and to reduce the cutting of tropical and temperate forests around the world. These options, however, may not fully reach their technical and economic potential due to various political and socioeconomic. The most practical of these is to increase CO2 sinks through photosynthesis in both standing tree biomass and in ocean primary producers. The use of marine algae as CO2 sinks is for large-scale CO2 mitigation: the use of phytoplankton through Fe fertilization and macro algal (kelp) farms, which can be used for energy production. The reduction of CO2-emissions that are damaging our climate is one of the major challenges of contemporary energy management. Nature itself offers us possibilities to produce energy CO2-neutral with the help of hydrogen producing micro-algae. Under certain conditions the light energy collected by photosynthesis is used to transfer electrons to hydrogen producing proteins called hydrogenases. A new type of hydrogenase that produces molecular hydrogen at relatively high rates was isolated (Happe and Naber, 1993) and characterized at the genomic level (Happe and Kaminski, 2002) for the first time in green algae. Processes were recently developed that allow long-term production of hydrogen by micro-algae (Melis et al., 2000). Under sulphur deprivation the green alga Chlamydomonas Reinhardtii adapts its metabolism from oxygen production and CO2-fixation towards hydrogen production. Therefore the biotechnological process is divided into the growth phase, the hydrogen production phase, and the resulting spent algae can be used for biomass production of fuel. Thus by control of growth conditions green algae can be used to produced hydrogen fuel, methane and a whole host of hydro carbons for fuel. Our planet is 75% ocean; it seems only natural to look to the ocean as a source of energy and a source to help lower our atmospheric CO2. The most appropriate regions for this kind of production would not only be the continental shelf regions but also in the open ocean where iron fertilization is being utilized to help with algal blooms. Algae and kelp may not be the holy grail of green energy sources, but they are a viable resource to help in our energy and environmental crisis.
Making Soap From Readily Available Agricultural and Household Wastes can Increase Cleanless in Rural Area
Eugenia Lucas and Thomas Ciaglo
In some areas of the world, soap is too expensive for many people to afford. For these people an alternative exists. They can make their own soap. In general, soap is made by the reaction of triglycerides and caustic soda. However, caustic soda, too, may be difficult to find or too expensive. The aim of this project is to develop a process for making soap from readily available agricultural and household waste materials, and other inexpensive chemicals. By using this process, rural people can get the benefits of readily available, inexpensive soap. Soap is made from animal fats or vegetable oils by saponification using strong base. The simple soaps can be isolated as cakes or bars, or it can be used as water solution. Many reaction conditions were studied to develop a recommended process that can be done using equipment and reaction conditions that can be performed in a kitchen or a fireplace. The soaps from this project were characterized primarily using infrared spectroscopy and several other analytical techniques as well as tests to show their effectiveness.