APEX Presentation Archives - 2012-13
March 28, 2013
My Life as a Beech Bum: Levels of Clonality in American Beech (Fagus grandifolia)
in Response to Habitat Disturbance
Academic Advisor: Dr. Nick Hirsch
APEX Advisor: Dr. Matt Hils
Location: James H. Barrow Field Station, Hiram College
Funding: Paul and Maxine Frohring Foundation gift to the James H. Barrow Field Station
American beech (Fagus grandifolia) propagates sexually through seed formation and asexually through root sprouting. Previous studies indicated beech populations may have more clonality with greater levels of environmental disturbance. Published data coupled with observations of beech-maple forest at the James H. Barrow Field Station in Portage County, Ohio suggest that habitat disturbance is associated with greater root sprouting. Thus, populations in disturbed areas would be expected to contain a higher proportion of clones than those in areas of minimal habitat disturbance. This forest consists of three main areas: a largely disturbance free mature area, an area of former pasture, and an area formerly containing chestnut trees decimated by chestnut blight. Comparison of the proportion of clonality among these three areas should provide insight into the relationship between habitat disturbance and levels of clonality. In order to determine the clonal identity of individuals, seven Simple Sequence Repeat (SSR) markers were used to assess the genotype of individuals. Individuals with identical genotypes were considered to be clones, while individuals with unique genotypes were considered to result from sexual reproduction. Preliminary results do not support much difference in clonality between plots sampled in the former pasture and in the mature forest areas.
A Comparative Locomotor Skeletal Analysis of the Order Lagomorpha
Major: Biology, Biochemistry
Academic Advisor: Dr. Matt Hils, Dr. Jody Modarelli
APEX Advisor: Dr. Sandra Madar
Location: Northeast Ohio Medical University
Supervisor: Dr. Jesse Young, Assistant Professor, Jesse W. Young Department of
Anatomy and Neurobiology, Northeast Ohio Medical University, Rootstown, OH
The order Lagomorpha contains two families, Ochotonidae (pikas) and Leporidae (hares/rabbits) that diverged 23 to 32 million years ago. Within Leporidae, the two genera being studied here, Sylvilagus and Lepus, diverged about 10 million years ago. Each divergence is marked by a differentiation in locomotor behavior that can be measured by examining skeletal structure. One measure that is particularly useful is mechanical advantage (MA) because it allows for the analysis of the force put into a lever system (i.e., the limbs) related to the force produced by the system. Mechanical advantage is measured by comparing lengths of the bones that constitute a joint of interest. Usually, high MA indicates a lever optimized for force production, whereas low MA indicates a lever optimized for speed. It was expected that the faster animal of interest, in this case Lepus, would have a lower average MA while the smallest, slower animal of interest, Ochotona, would have the highest with Sylvilagus being an intermediate. Once measurements were taken and tabulated, Ochotona always had the highest MA for each joint and matched the expected results while Lepus and Sylvilagus did not meet expected results as Sylvilagus had lower MA than Lepus for some measurements.
Protein Analysis of Mineralization in the Gastrocnemius Tendon (Achilles)
of the Domestic Turkey
Academic Advisor: Jody Modarelli
Location: The University of Akron Department of Polymer Science
Supervisor: Dr. William J. Landis, G. Stafford Whitby Chair in Polymer Science,
Department of Polymer Science, University of Akron, Akron, OH,44301, Phone:
Source of Funding: The University of Akron Department of Polymer Science,
The McNair Scholars Program (TRiO)
Mineralization occurs in various tissues, including: bone, enamel, cartilage, and tendon. Physiologically, mineral formation allows for increased load-bearing and protection from injury. Chemically, mineralization occurs from formation of hydroxyapatite crystals in the extra-cellular matrix (ECM). However, in certain instances the genesis of calcium deposits in connective tissues can be harmful to the overall well-being of the individual; such is the case with calcific tendonitis of the rotator cuff. The etiology of why calcium builds up in some areas but remains absent in others is unknown. An interesting model to study is the gastrocnemius (Achilles tendon) of the domestic turkey, Meleagris gallopav, due to its unique pattern of mineralization. Three theories have been proposed as to the nature of this phenomenon. They are: the presence or absence of vasculature networks, protein content, and mechanical stressors. It is the aim of this study to demonstrate that proteins are the underlying cause, specifically, small, non-collagenous proteins of the ECM. We have identified significant variations in biglycan and bone-sialoprotein and suggest that they are most likely involved in mineralization of tendons. Further study is necessary to determine how these proteins interact with the ECM and collagen fibrils to form mineral.
Edge Effects in a South-Facing Beech-Maple Successional Forest in North-Eastern Ohio
Academic Advisor: Professor Matt Hils
APEX Advisor: Professor Michael Benedict
Location: Hiram College James H. Barrow Field Station
Source of Funding: Frohring Foundation
Forest edges create microclimate conditions that influence the structure and composition of plant communities. Such edge effects have high ecological and conservation relevance because they decrease functional forest-interior areas. Plant species richness is higher at edges of forests and may promote increased abundance of exotic invasive species. Edge effects are more pronounced on south-facing edges in the northern hemisphere because of greater sunlight exposure.
The James H. Barrow Field Station (Hiram College) contains one of the few remaining old-growth beech-maple forests in the state of Ohio and is managed for its protection. Considering this site's importance, and in support of its conservation management, this ongoing study investigated the extent of edge effects in a south-facing successional beech-maple forest by quantifying environmental parameters and compositional structure of woody and herbaceous vegetation along an edge-to-interior gradient.
It was hypothesized that light, wind speed, air and soil temperature would decrease while air humidity and soil moisture would increase into the forest. It was hypothesized that plant communities were suggested to respond to the altered environmental parameters. Preliminary results support these hypotheses with the exception of wind speed, which was noted as having no overall pattern.
March 13, 2013
Model Reed Bed System for the Filtration of water in order to reducethe influence of protozoa in regards to Mycobacterium Avium
Academic Advisor: Dr. Matt Hils, Department of Biology
Supervisor Dr. Jody Modarelli
Source of Funding: Paul and Maxine Frohring Foundation
Abstract: Conservation efforts by zoos are being made to repopulate the endangered White-winged Wood Duck (WWWD) in captivity, a species highly susceptibility to Mycobacterium Avium (MA), the causative agent of Avium tuberculosis (TB). Changes in the environment can alter water and soil conditions by bringing in foreign debri and non-native organisms like protozoa. Protozoa have been shown to harbor MA and facilitated replication, which would put waterfall at risk that have ingested protozoa. The purpose of this study was to construct a model subsurface horizontal flow reed bed with different substrates to assess whether a reed bed system is effective in reducing or eliminating protozoa found in the water where ducks dabble. Water was collected at the Akron Zoo from the duck deep pool and filtered through the common reed, phragmites australis, in a susbtrate of either gravel or sand/gravel. Samples from unfiltered and filtered water was plated at 6 hours, 12 hours, and 24 hours. Colonies were grown in a CO₂ incubator at 40˚C and 5% CO₂ for approximately four weeks before being extracted and quantified. Preliminary results show that reed plus the sand/gravel substrate was the most effective in reducing the amount of protozoa in water.
Polymer Synthesis: A Biochemical Approach
Academic Advisor: Dr. Jody Modarelli, Department of Chemistry
Location:University of Akron, Department of Polymer Science
Source of Funding: Omnova
Abstract: Polymers are used to produce materials that are essential to both enjoying and sustaining life. However, due to the significant amounts of materials made from polymers, disposal has become a real concern. In order to explore the potential of utilizing naturally occurring chemicals and biodegradability in industrial applications, D,L-serine was investigated as the initial reagent involved in the synthesis of a (2-bromo-2-butoxycarbonyl)ethyl acrylate inimer. The methods employed included a diazotization reaction of D,L-serine, which substituted the amine group with a bromine group to yield 2-bromo-3-hydroxypropionic acid, which was then esterified with butanol to yield butyl 2-bromo-3-hydroxypropionate. In the final step of the reaction, butyl 2-bromo-3-hydroxypropionate was reacted with acrylic anhydride to yield the (2-bromo-2-butoxycarbonyl)ethyl acrylate inimer. This inimer will be further studied to ensure it can be polymerized into similar synthetic industrial polymers by utilizing polymerization and analytical techniques. Furthermore, the degradation of the polymer will be attempted to ensure biodegradability.
Detection of Cholesterol Esters in Patients with Renal Cell Carcinoma
Academic Advisor: Dr. Jody Modarelli, Department of Chemistry
Source of Funding: Paul and Maxine Frohring Foundation and the National Science Foundation: Major Research Instrumentation grant # 1039259
Abstract: Renal Cell Carcinoma (RCC) characterized by cancerous cells that line the tubules of the kidney accounts for approximately 70% of all malignant kidney tumors. It is referred to as the "silent disease" because often spreads beyond the kidney without symptoms. Prior studies have shown that cholesterol and cholesterol esters (CE) play a significant role in cell proliferation. Healthy kidney tissue consists of high amounts of free cholesterol, whereas RCC tumors contain high amounts of cholesterols in their esterified states, mainly as oleate. The aim of this study was to assess whether cholesterol esters from RCC tumors are shed into the urine. Cholesterol esters from the urine of control and RCC patients were extracted using a modified Bligh Dyer, separated by Thin-Layer Chromatography (TLC) and analyzed by Electro-Spray Ionization Mass-spectrometry (ESI-MS). The underlying goal is to develop a non-invasive, Enzyme-Linked Immunosorbent assay (ELISA) for in order to detect RCC in early-stages. To date, evaluation of twenty cholesterol esters, including oleate at m/z 302, does not support the hypothesis that cholesterol esters are shed into the urine of patients with RCC. It is possible that this is due to a modification of the esters prior to being shed in the urine.
March 20, 2013
Decomposition rate and macroinvertebrate colonization of artificial leaf packs in a forested headwater stream
Academic Advisor: Jody Modarelli
APEX Advisor: Jennifer Clark
Location: Hiram College/Hiram College Field Station
Abstract: Leaf packs are natural sources of nutritional input as well as habitat for invertebrates in stream ecosystems, and can vary widely in composition and colonization by organisms. In this study, we examined leaf decomposition rate and macroinvertebrate colonization using three different types of artificial leaf packs (American Beech, Sugar Maple, and a combination) in riffles (n = 3) and pools in Silver Creek. Subsets of the leaf packs were removed two, four, and six weeks after initial deployment during winter 2012 and a variety of abiotic parameters were measured. The dry weight of leaf species significantly decreased over the three sampling periods (P < 0.0001) with Sugar Maple having significantly more mass loss than the combination treatment and the combination treatment having significantly more mass loss than American beech (P < 0.0001). However, the effect of habitat type on leaf decomposition was not significantly different between riffles and pools (P = 0.9010). Further, macroinvertebrate abundance significantly increased throughout the three sampling periods (P < 0.05) but there was no significant difference between leaf types (P = 0.4165). These results suggest that the biotic community instead of abiotic parameters play a strong role in leaf decomposition in system.
The James H. Barrow Field Station Bird Guide
Academic & APEX advisor: Sandra Madar
Location: James H. Barrow Field Station
Supervisor: Laura Collins, Previous Associate Director of the James H. Barrow Field Station
Funded through the James H. Barrow Field Station
Abstract: The James H. Barrow Field Station Bird Guide was created during the summer of 2012 as an educational resource for a variety of uses. The Guide is a collection of 41 bird species that are common to the greater Ohio area, illustrated with pencil and ink. Paragraph sized species descriptions were also written to pair with each image. The illustrations have been photographed and are currently being created into booklets with Photoshop and Microsoft Publisher applications. They will be printed as necessary and uploaded to the Hiram College webpage as an online resource for Field Station visitors.
Using Classical Conditioning to illicit learned changes in directional response to stimuli in terrestrial isopods
Academic Advisor: Nicholas Hirsch
APEX Advisor: Jennifer Clark
Abstract: The ability of an organism to adapt to abrupt environmental changes is often used as a mark of intelligence in ethological studies. Previous studies have shown isopod behavior to be mostly mechanical and instinctual but not a function of higher associative learning. This study investigated whether Armadillidium vulgare, a terrestrial isopod species possessing basic CNS and nerve ganglia, could learn to change its taxis to a variety of stimuli through classical conditioning. The directional response to several categories of stimuli (magnetism, humidity, light frequency, light intensity and several food sources) was divided into taxis and unconditioned responses based on the distribution frequency of isopod visits within a Y-maze. Classical conditioning was used to train isopods to form associations between unconditioned and taxis stimuli. Results indicated a gradual increase in isopod visits to the unconditioned stimulus chamber after each trial and by the end of 4 training trials the frequency of visits of isopods to an unconditioned stimulus resembled the frequency of a taxis response up to 2 days after the removal of the taxis stimuli before reverting innate behavior. This suggests that the isopod behavior may not be entirely governed by instinct but demonstrate a limited period of learning.
The Homeotic Gene Pax-6 and Its Role in Hindbrain Development
Academic & APEX Advisor: Nick Hirsch
APEX Advisor (meaning Hiram person): Nick Hirsch
Abstract: The goal of this project was to determine the temporal and spatial expression of Pax6 genes in early X.tropicalis (frog) embryos. Pax6 is a key factor that enables developing embryonic cells to differentiate into neural cells that give rise to specific regions of the brain. Mutation of this gene in humans has been shown to result in small or missing eyes (aniridia), and may also contribute to other brain malformations. The focus of our study was the hindbrain, an evolutionarily ancient part of the brain that is very similar in frogs and humans. The hindbrain is composed of the cerebellum, the pons and the medulla oblongata. The cerebellum is important in regulating movement, while the medulla and pons control essential unconscious functions such as breathing, heart rate and balance. Some human disorders linked to abnormal hindbrain development include ataxias (a family of balance disorders) and some forms of autism.
Validation of Fecal Glucocorticoid Metabolite Assays in White Winged Wood Ducks
Academic & APEX Advisor: Dr. Jody Modarelli, Department of Chemistry
Location: The Akron Zoo
Funding: Paul and Maxine Frohring Foundation
Abstract: The endangered White Winged Wood Ducks (WWWD) are facing threats in captivity due to mycobacterium avium, the causative agent of Avian Tuberculosis. Waterfowl are more susceptible to disease when their immune systems are lowered due to stress. The purpose of this study was to monitor and validate the stress hormone levels of the WWWD in captivity using an Enzyme Linked Immunosorbent assay (ELISA). Stress hormone levels are important to monitor because they could be indicative of a lowered immune system, thereby making them more susceptible to disease. Fecal samples were collected from the captive population at the James H. Barrow Field Station in order to test for fecal glucocorticoid metabolites, which contain stress hormones that can be monitored non-invasively. Samples were collected for two weeks, which induced stress, and then taken for another two weeks post-stress. All samples were lyophilized prior to the ELISA assay, which was performed in collaboration with the Cleveland Metropark Zoo. We hypothesize that stress hormone levels will peak after a stressor and will decrease and return back to normal post-stress. Future directions include monitoring other populations of WWWD and other endangered waterfowl with the goal of offsetting disease in endangered species.
November 14, 2012
Comparing the Success Rates of Artificial Insemination and Natural Service in Cattle
Academic and APEX Advisor: Sandy Madar
Supervisor: Randy Alger, DVM; large-animal veterinarian; 3413 Mennonite Road Mantua, OH 44255; (330) 274-3463
Efficiency of reproduction is one of the most critical aspects determining profitability of a cattle herd. Currently, both artificial insemination and natural service are common methods of fertilization. However, many aspects must be analyzed when determining the most suitable method. The purpose of this study was to determine which method is more universally practiced in the cattle industry, and how these related to economic impacts, safety, complications, and success rates. The implications for the assisting veterinarians are also examined. Results indicate that artificial insemination is a more economically sound decision for both the farmer and clinician.
Total Knee Replacements
Academic and APEX Advisor: Nick Hirsch
I did an internship during the summer of 2012 at Dr. Donald L. Fisher, MD Inc. orthopedic physical therapy clinic. While there, I became interested in the factors affecting total knee replacement (TKA) rehabilitation. Previous research has shown that males and females show no significant differences in the amount of functional range of motion and overall improvements gained post-surgical repair. Different physical therapy techniques are frequently used in rehabbing a TKA patient, but they all generally lead to the same end results. Using an anonymous sample of 60 patients (30 male, 30 female) who underwent TKA repair by Dr. Fisher, I compared the amount of flexion and extension gained after sufficient physical therapy versus the patient’s sex and age. The results show that there are no significant differences between sexes and the amount of range of motion gained, but increasing age does show a slight decline in motion recovered. These results are agreeable with the current literature. Future studies may look at the effect of weight on TKA motion recovery as well as the amount of impairment and joint corrosion a patient has pre-TKA.
Reducing the number of concussions in sports: Physically and Mentally
Academic & APEX Advisor: Sandy Madar
Over the past ten years there has been a dramatic rise in sports related concussions. There is an estimated 4 million concussions occurring each year due to physical activity resulting from sport participation. Recent studies indicate that high impact sports such as American football, hockey, and lacrosse have the highest concussion rate amongst active sports. Data analysis show female athletes and younger children participating in high contact sports are more prone to concussions. Female athletes have a longer recovery period when treated for post-concussion symptoms compared to male athletes. Biomechanical studies of concussions can measure the acceleration of the head post-impact. These studies have shown that the magnitude, distribution of magnitude and frequency of the impact all play a role in the severity of a concussion. Strengthening neck and core muscles decrease the acceleration of the head after a traumatic impact and reduce the effects of a concussion. How quickly an athlete can contract the muscles before sustaining a hit is a factor in injury probability. This research shows the importance of physical and mental preparation in preventing concussions in young athletes. There are many other unknown factors that contribute to traumatic head injuries in high-impact sports that need further investigation.
The Exploration of Chronic Pain Assessments through Research
Academic and APEX Advisor: Sandy Madar
The Numerical Rating Scale and McGill Pain Questionnaire are tools used in research and medicine, which help assist those who are struggling with pain. Pain is the primary reason why individuals seek medical attention because of the impact it has on their daily lives. Successful treatment plans require determining which pain management technique is most appropriate. Until recently, the primary tools for clinicians were pharmaceutical; now however, there is a growing industry developing devices that can aid in pain management. Once such company is Neuros Medical, Inc. Accordingly, while interning with Neuros, assessments such as the McGill Questionnaire were used to identify the central cause of pain and to measure its impact on patients’ lives. Additionally Neuros Medical uses such tool to measure whether its device is both safe and significantly effective in improving patient pain levels, an important step in gaining FDA approval.
Fallen Feathers: A Review of the Implications of Imprinting in Raptorsand the Necessary Precautions for Rehabilitators
Major: Biology and Environmental Studies
Academic Advisor: Denny Taylor and Sarah Mabey
APEX Advisor: Denny Taylor
Source of Funding: Donations only
Imprinting is the process of learning during a sensitive period of development in an animal’s life that results in a particular behavior pattern that is independent of the consequences of behavior. In raptors and many precocial birds, imprinting occurs early and results in acquiring normal behavioral characteristics from parents. Although imprinting occurs at different levels, a raptor fully imprinted on something other than a raptor parent suffers the most detrimental form of mental instability and faces several problems later in life, such as the inability to breed with members of its own species and the inability to recognize threats. Research on imprinting is sparse, especially on ways to prevent unintended imprinting, specifically in rehabilitation centers. Presented is an overview of when, why, and how imprinting occurs in birds of prey, as well as a summary of the current methods used in rehabilitation centers to prevent deleterious imprinting, with a call for further research and publication on ways to prevent unintended imprinting in rehabilitation centers.
APEX Presentation Archives - 2011-12
November 9, 2011
Sea Turtle Nesting Program: How to Turtle Walk
Sea Turtles play a major role in the history and culture of the Cayman Islands, but overfishing during the 17-1800's has left most species of sea turtles endangered, or critically so. The Cayman Islands Department of Environment has been working to restore the numbers of sea turtles in the Caribbean by running a nesting conservation program since 1999. Three species of sea turtles: Green (Chelonia mydas), Loggerhead (Caretta caretta) and Hawksbill (Eretmochelys imbricata) are monitored and protected by the Department of Environment's Sea Turtle Nesting Program. The program works by monitoring the nesting activities and hatch rates for these three species of sea turtles on Grand Cayman's coastline. The number of turtle nests has increased significantly since the program has begun (from 21 nests in 1999 to 240 nests in 2010). The beaches are walked twice a week, and the location of each nest is recorded. After the nest has hatched the egg chamber is excavated and the hatch rate calculated. As the methods used in the program have not changed since the program began it appears that the critically endangered sea turtle nesting populations in the Cayman Islands are recovering, as nesting numbers continue to increase.
Using Metagenomics to Understand Microbial Communities
A metagenome includes the DNA of all organisms, usually microbes, present in a particular habitat. Its major benefit is to illuminate the presence of unculturable bacteria. Two of the projects I was involved in during the summer of 2011 were concerned with the microbial community on the leaves of maize. This habitat, called the phyllosphere, is quite harsh – fairly dry, exposed to lots of UV light, and potentially low nutrients. To what extent do these conditions influence the phyllosphere microbial community? One way to answer this question is to compare communities growing on different genetic varieties of maize.
Our lab collaborated with Stuart Gordon at Presyberterian College, and the OARDC to grow 25 different lines of maize. Metagenomic data from a planting of Missouri 17 maize in 2009 was analyzed based on 16S rRNA gene sequences from these samples. The 16S data was sorted into "clusters" based on matches at the level of genus and order using a computer software package developed at Hiram College. The results so far indicate that the phyllosphere, like microbial communities in almost all other habitats, is primarily dominated by a few types of microbes with most of the diversity comprised of rare taxa.
The Blue Iguana Recovery Program: Health Assessments 2011
The Blue Iguana Recovery Program of the Cayman Islands was established 20 years ago on Grand Cayman. The top priority of the program is the conservation and captive breeding of the species Cyclura lewisi or the Grand Cayman Blue Iguana, endemic to the small island of Grand Cayman. This beautiful and majestic creature was at one point in time near extinction and is currently on the endangered species list. With the goal of the program being to grow the population to 1000 blue iguanas in the wild, nest excavations and egg incubations are carried out every year as an important part of the recovery program. Every year, there are health screenings/assessments carried out on all of the pre-release 2 year old captive bred individuals and the captive breeders. Once health screenings have been carried out, the 2 year old individuals are then released into protected reserves on the island.
A Tale of Two Bees: Nesting and Foraging Behaviors of Solitary Bees
The adult foraging behaviors of two solitary bees, Dufourea calochorti (Halictidae) and an unnamed Osmia sp. (Megachilidae) were observed in the Western Cascades of Oregon. Osmia sp. was determined to be a ground-nesting species, a habit generally considered under-represented in the genus. Fifteen nests were excavated to determine burrow architecture and cell structure. Burrows proved to be diverse in structure, possibly due to poor soil conditions. Adult wasps of the family Chrysididae, known kleptoparasites of many other hymenopteran species, were seen entering nests. Cells proved to be similar to those of other ground-nesting Osmia spp. in North America. Twelve capped cells were dissected to determine their contents, revealing evidence of kleptoparasitism by Stelis sp. and Chrysidid wasps.
November 16, 2011
Impact of Plant Nutritional Status on the Phyllosphere Bacterial Community in Zea mays
The phyllosphere is defined as the surface area of a leaf that interfaces with the atmosphere. These surfaces represent the largest biological interface on Earth and are home to many bacteria, fungi and other microorganisms. These microbial communities spend their lives in a microscopic landscape consistently bathed in ultraviolet radiation, subjected to periods of drought and periods of deluge, and nearly barren of nutrients, and yet they not only survive, they proliferate. This study used metagenomic techniques to investigate this unique microbial community. This molecular culture-independent approach allows a much more comprehensive view of what organisms exist in a given environment as only a tiny fraction (<1%) of environmentally sampled bacteria will grow in the laboratory with current culture techniques. Maize was chosen as a plant model for this study due to its relevant agricultural importance and because it is widely studied and easily grown. Two in-bred lines of maize commonly grown for agriculture in the U.S. were grown under differential treatments in an outdoor setting to answer the questions: What, if any, effect does a plant's nutritional/health allowance have on its community of phyllosphere microbes and can these microbial populations be indicative of a plant's health or stress level?
Hibernation's effect on bone strength and density in Marmota monax
The woodchuck (Marmota monax) is the largest true hibernating animal. Woodchucks hibernate up to eight months per year, consuming no nutrients. In addition to a drastic decrease in metabolism, woodchucks also greatly reduce core temperature during hibernation. Despite being completely inactive for several months, woodchucks, a prey species, must be physically able to evade predators upon the termination of hibernation. Bone strength and density is normally regulated hormonally through calcitonin and parathyroid hormone (PTH). PTH is a hormone that would typically release calcium from bone to increase blood calcium. Because woodchucks consume no nutrients during hibernation, it would be assumed that PTH would regularly release calcium into the bloodstream to maintain appropriate blood calcium levels. This would in turn cause osteoporosis in the woodchuck. Woodchucks were studied at either hibernation, pre-hibernation, post-hibernation, or summer stages. X-ray measurements were taken on long bones, and micro-CT scans analyzed bone density of the metaphyses and diaphyses of the harvested long bones. An Instron machine was used to determine bone strength of harvested long bone through three point bending. These tests displayed no change in bone strength or density between activity stages, and juvenile animals' bones continued to grow even in hibernation.
Isolation of Agrobacterium tumefaciens strains for soybean transformation
Soybeans are highly valued crops and are a primary target for genetic transformation. Agrobacterium mediated transformation is one method transforming many plants such as corn and soybeans. Unfortunately both the strains of Agrobacterium tumefaciens and the techniques used are inefficient. This paper describes the first part of a larger project in which new strains of A. tumefaciens are isolated and tested for their ability to transform soybeans. A. tumefaciens strains were gathered from both soil samples of soybean fields and crown galls of a variety of plants. DNA was then isolated from the bacteria using the heat shock method and run in PCR targeting the virulence gene VirG. PCR products were run on a 1.5% agarose gel. VirG has a base pair size of 539and positive results were determined with an amplification of this size. Research so far has resulted a few possible strains that contain VirG and more is expected over time.
A Mouse Model of Testicular Germ Cell Tumors and Spontaneous Metastases
Over the past 30 years there has been a worldwide trend of testicular cancer affecting younger and younger men, as well as more men overall. While the cause for this is unknown, genetic risk factors are being discovered. The 129 strain of inbred mice are ideal for researching this disease because they are prone to developing testicular germ cell tumors, which account for the majority of testicular cancer cases in men. For the first time, spontaneous metastases developing from primary germ cell tumors have been identified in this strain. Necropsies were conducted and the number of tumors, metastases and their locations within the body were recorded. As in human cases, metastases were most commonly found on the lymph nodes. Using these mice as a model, we can improve our understanding of the formation and progression of testicular cancer.
Upper Cervical Chiropractic Treatment for Fibromyalgia
This paper describes the muscular condition fibromyalgia (FMS) and how upper cervical chiropractic can be used to treat this disease by getting the atlas adjusted to relieve pressure off of the nerves flowing through the spinal cord. The upper cervical technique described in this article is NUCCA (National Upper Cervical Chiropractic Association). A review of research done on patients with fibromyalgia shows the success of adjustments, or spinal realignment by a chiropractor, in decreasing debilitating symptoms.
March 21, 2012
Identification of Tannins Found in Mast from Tree Species found on the James H Barrow Field Station
Tannins are a plant secondary compound that prevents herbivore predation by reducing the digestibility of proteins. Many species of nuts contain tannins, and many animal species eat these nuts despite the presence of tannins, including the eastern grey squirrel. The types of tannins in five nut species were identified though mass
Evaluating and Restoring Wetlands
The initial goals of this internship were to locate, determine, evaluate and delineate the wetlands at the James H. Barrow Field Station (JHBFS) belonging to Hiram College. Based on this data, two or more wetlands were to be constructed on the JHBFS property for the purposes of research and education. The following is a report of the steps of this multilevel project that employed multiple interns. Included in this project was a service learning program, an education program, and the creation of a website and a social media presence to encourage a relationship with the surrounding community. Records of wetlands and wetland research at the JHBFS were standardized and organized. Procedures included but were not limited to GPS mapping, determination, ORAM use, construction planning and elevation mapping. At the end of the internship 39 wetlands were determined and located and 3 wetlands were designed for different locations. Frohring stewardship interns gained skills in Garmin and Trimble GPS use, GIS use, navigation, elevation mapping, map making, plant identification, delineation, ORAM use, external resource collaboration and construction planning. Connections were built between collaborators, Nitisha Shrestha, Diane Walcott, Sonia Bingham, Joel Bingham and the Cuyahoga Soils and Water Conservation District.
A Unique Micellular Vesicle Found in the Endangered White Winged Wood Duck
Conservation efforts by zoos and protected breeding grounds are being made to repopulate the endangered White-winged Wood Duck (WWWD) in captivity, but have met with resistance due to their high susceptibility to Mycobacterium Avium (MA), the causative agent of Avium tuberculosis (TB). Birds living in a soil-water environment are highly susceptible to MA due to the lipids in its cell envelope that form a biofilm. In this study, we use Liquid Chromatography Mass Spectrometry (LCMS) to identify a unique set of lipids in the plasma of our ducks, despite their living conditions or exposure to MA ranging from m/z 902 to 1,581. These lipids spontaneously formed a unique reverse micellular vesicle. From our MS/MS data, we found that this group of lipids corresponds to a species of diacylglycerols. With a hydrophobic tail on the outside and a hydrophilic tail on the inside, reverse micelles can accommodate small peptide metabolites. Along with the MA cell envelope, these types of reverse micelles may also act as a determinant of virulence and could contribute to the duck's host immune response to MA, making ducks highly susceptible to infection with MA upon contact with the biofilm in a soil-water environment.
Are You What You Eat? An Intake Study of Three Lemur Species' Diets to Evaluate Nutritional Values of Various Nutrients
The diets of three lemur species, black and white lemur (Varecia variegata), red ruffed lemur (Varecia variegata rubra), and ring-tailed lemur (Lemur catta), were studied to evaluate the nutrients being obtained in their diet in order to provide proper nutrition and prevent disease. Preparing a proper diet for captive animals can be challenging because the natural diet in the wild cannot be imitated in captivity. The diet in captivity must be prepared with local foods available in our area. Because of the possibility of poor nutrition in captivity, it is important to closely monitor their nutrition. The items in the diet were weighed for seven days. The amount of food consumed was entered into the computer program "Zootrition", which provided values of various nutrients obtained in their diet. In this study, we focused on iron, fiber and folacin. Results showed that the lemurs were consuming too much iron, not enough fiber, and not enough folacin. Based on this data, changes were made to their diet and the study repeated. Although improvements were made, our study showed that more changes are necessary in order that the lemurs can reach the proper nutritional values recommended.
March 29, 2012
Hindbrain Gene Expression and Rhombomere Patterning in Xenopus Tropicalis
With the field of developmental biology constantly advancing and new techniques becoming readily available, the development of the hindbrain has sparked new interest. It has been known that problems with hindbrain development and patterning can lead to various human disorders such as autism, but only recently has the genetic expression which may cause these disorders been studied. Recent research has shown that the segments in the hindbrain, called rhombomeres, each contain a specific regional identity and become fated for certain structures in the hindbrain. Through the expression of homeobox genes within the rhombomeres of the embryos, it has been found that rhombomere specific expression domains are established, leading to correct and proper hindbrain function in adults. In this study, the expression of otx2, a gene known for its role in hindbrain development in other species, was mapped in Xenopus tropicalis through in situ hybridization. In situ hybridization shows where otx2 genes are active in the embryo through staining.
Jemika Shrestha Kastee
Expression of clock genes in the brain of Xenopus tropicalis
The biological clock determines the timing of rhythms in behavior, physiology and biochemistry of organisms ranging from microorganisms to plants and animals. Some of the central clock genes like period1, period2, bmal and neuropeptides like vasoactive intestinal polypeptide (VIP) and arginine vasopressin (AVP) can be used to monitor changes in the circadian clock. In this research, these genes and neuropeptides were tracked in the Xenopus tropicalis brain using in situ hybridization. Brain sections were collected every 4 hours for 24 hours. After being probed with radioactively labeled antisense RNA, these sections were exposed to film and analyzed for spatial expression patterns. The results showed that AVP was highly expressed in the SCN and to a lesser degree in the hypothalamic preoptic area. VIP was highly expressed in the SCN, ventral thalamus, and ventral hypothalamus of the forebrain sections; bmal was found in different regions of the forebrain and in torus semi circularis and nuclei isthmi of midbrain. period2 was found to be highly expressed in olfactory bulb, lateral pallium, dorsal pallium, ventral striatum, thalamic axis, hypothalamic preoptic area, ventral hypothalamus. period1 was found in various regions of forebrain midbrain and in the cuneate nucleus of the hindbrain.
GLISTEN: Improving Local Watersheds through Citizen Science
The Great Lakes Innovative Stewardship through Education Network (GLISTEN) stewardship liaison program allows undergraduate scholars to develop and implement service-learning projects with peers at their universities. As a stewardship liaison (USL), students serve an important role in improving local watersheds by providing a link between community partners and their universities. Michelle Crowl and I as Hiram College USL partnered with the Hiram Historical Society, along with HC alumna and citizens to determine the needs and utilize local materials to rebuild hardscapes for a vegetable-herb garden at the Century House and for a historic knot garden at Bonney Castle. Hardscapes were built using local, organic materials. Interest regarding the gardens was generated through social media sites and a community outreach program was created and implemented using the plans and gardens created in this project. We then collaborated with the Frohring wetland interns determining, delineating, and classifying wetlands at the James H. Borrow Field Station. This knowledge was incorporated into a service-learning based course curriculum, Introduction to Field Biology, a non-majors course at Hiram College. A summary of this program was presented on Capitol Hill to members of congress and their representatives in an effort to demonstrate how working on such a small-scale can have a large-scale impact.
Iroquois 1 Gene Expression in the Xenopus Hindbrain
Malformations of the hindbrain are thought to cause disorders such as autism and ataxia, amongst others. Many genes are active in the process of hindbrain formation, but the gene discussed in depth in this talk will be Iroquois1 (Iro1). This gene works to determine cell fates of neural cells during the process of gastrulation and neural crest cells during neurulation. In this experiment, probes were created using a variety of microbiology techniques and these probes were used to perform in situ hybridization on Xenopus tropicalis embryos in order to determine where in the developing neural tissue the Iro1 gene was being expressed.
April 4, 2012
Tuning the Tools: Establishing Proof of Concept and Methodology for a Botanical Study
The purpose of this project was to establish proof of concept for a genetics-based population study of American beech, Fagus grandifolia. This native tree species can reproduce either sexually, through seed production, or clonally, by growing sucker sprouts directly from its roots. Research and lab work focused on extracting and analyzing DNA from leaf tissue to determine whether a tree is a clone or a seedling. DNA extractions were performed on both silica-dried and frozen leaves to determine which method best yielded DNA samples. The extraction protocol itself was tested and reevaluated until it yielded proper product. PCR reactions were tested to amplify 10 loci per sample, and these products were sequenced to determine genetic identity. With the establishment of this method and its protocol it is possible to compare genetic data with the sampling and distribution data. It is then possible to address population and environmental questions relating to beech's reproduction methods; and whether or not certain conditions cause it to favor clonal or sexual reproduction in a given circumstance.
New Experiments on Bacterial "Charity"
A recent paper by Lee et al (Nature 467:82-5) has shown a population-level signaling mechanism, acting in concert with traditional natural selection, can contribute to bacterial antibiotic resistance. Rare truly-resistant E. coli can secrete the chemical indole, which helps their nonresistant neighbors to survive exposure to antibiotics that should normally kill them. The indole induces changes in gene expression that increases survivorship under stressful conditions. I conducted a set of experiments to determine whether a similar mechanism would be seen when different antibiotics were used, and whether it only occurred in bacteria capable of indole production. Kirby-Bauer antibiotic testing was used in a protocol adapted from Weldon and Hossler (Am. Biol. Teacher 65:56-61). E. coli was selected for testing because of the previously demonstrated resistance patterns and indole signaling, along with Agrobacterium tumefaciens due to its inability to produce indole. Four rounds of testing were carried out with both strains in combination with four different antibiotics. The results showed similar resistance patterns for both bacteria under all four testing conditions.
Does Yogic Breathing Reduce Heart Rate More Effectively After Being Startled?
Yogic breathing, when practiced regularly has been shown to reduce heart rate. Volunteers who practice yoga were recruited and asked to participate in a study to demonstrate the technique's effectiveness. Each subject's resting heart rate was taken, they were then startled, and the time it took each subject to go back to resting heart rate was recorded. Two groups of subjects were tested, one group who knows the technique and another who does not. Preliminary results show that there is no significant difference in recovery time between the two groups, disproving the hypothesis that yogic breathing reduces heart rate more effectively.
TILLING: A Novel Method to Quickly Identify Mutations
In recent years, reverse genetics approaches have emerged that are interested in studying genetic variation and gene function. Targeting Induced Local Lesions In Genomes (TILLING) is one such approach that allows researchers to locate single nucleotide mutations in a variety of organisms. Mass spectrometry is one technique used in TILLING which allows the detection of these mutations within nucleotides. Corresponding with Hiram's recent purchase of an LC-ESI-MS, interest has formed regarding the use of this equipment to study mutations in the Xenopus tropicalis gene Pax 6, a master regulator of eye development. In this experiment, sets of oligodeoxynucleotides were designed where one of the oligodeoxynucleotides contained a single base pair change. These samples were directly infused into the ESI-MS and analyzed with hopes to detect the difference in mass between the samples. While a difference in the mass to charge ratio was observed, it is clear that more information is needed to optimize this technique before any future experiments begin.