Journal of Biological Research, Vol. 9 (2008)

Skin Gland Regeneration in the Red-Backed Salamander, Plethodon cinereus. NATHALIE AALL, Biology Department, Hartwick College. The purpose of this study was to compare skin gland regeneration in dorsal versus ventral skin of regenerating tails of Red-Backed Salamanders. These salamanders use their tails in territorial marking and courtship by dragging the ventral surface on the substrate. I hypothesized that the glandular differences between dorsal and ventral skin is due to the ventral skin being exposed to more friction than the dorsal skin. If this is true, then there should be fewer differences between dorsal and ventral skin glands in newly regenerated skin than in mature skin. To test this idea, amputated tails were harvested at regular intervals over a period of thirteen weeks and analyzed histologically. Selected samples were also labeled with BrdU to examine localized mitotic activity using immunocytochemistry. Faculty Supervisor: Dr. Stanley Sessions.

Conservation of Osiris genes in diverse arthropod species. ALEXIS D. ANDRUS, Department of Biology, Hartwick College, Oneonta, NY 13820. The Osiris gene family is a cluster of genes found to encode a novel family of proteins in insects. In this study we will look at Osiris gene conservation. We hope to discover the Osiris genes’ evolutionary origins and divergence patterns. In addition, we would like to find when their orthologous nature developed within this diverging process. In order to unearth the evolutionary beginnings of Osiris genes we created degenerate primers which will hopefully amplify transmembrane domains in a variety of insect species, beginning with different Drosophila species. There is also the possibility of amplifying these domains in individuals of other classes closely related to the class Insecta. When and if these degenerate PCR reactions yield products they will then be sent for sequencing, and in turn sequences will be analyzed for similarity to known genome sequences using BLAST. By determining which species contain Osiris genes we will be able to determine the molecular relationships between taxa and obtain a clearer picture of the phylogenetic history of Osiris genes. By investigating species closely related to those in the class Insecta we hope to determine whether or not Osiris genes are unique to insects. Faculty Supervisor: Douglas Dorer

Localization of the expression of PAB3 and PAB5 proteins in plants using the Green Fluorescent Protein. SHANE CHERICO, Department of Biology, Hartwick College, Oneonta, NY 13820. In eukaryotes, Poly(A) Binding Proteins (PABP) attach to and modify mRNA as it is transcribed and exported from the nucleus for translation (Chekanova, et al., 2001). For plants, the deletion of certain PABP leads to a lethal phenotype. Most eukaryotes have only one or a few PABP’s, but eight expressed PABP genes can be found in Arabidopsis thaliana. Various Arabidopsis PABP’s are differentially expressed, but two are known to be pollen specific: PABP3 and PABP5 (Belostotsky, 2003). To identify in which pollen cell these proteins are expressed PCR was used to amplify the PABP3 and PABP5 promoter regions, which were then ligated separately to the GFP coding region attached to a nuclear localizing signal. Gateway cloning was used to put the promoter/coding region construct into a binary plasmid. In the future this plasmid will be used in plant trangene experiments. Faculty Supervisor: Dr. Douglas Hamilton.

The effect of crowding on Amphibian deformities. MELISSA CHRYSOSTOME, Department of Biology, Hartwick College, Oneonta, NY 13820.The occurrence of deformities in natural populations of amphibians has emerged as one of the most important environmental issues of the last decade. Some deformities (especially extra limbs) are now known to be caused by parasites (trematodes). In other kinds of deformities (especially missing limbs), the causes have not yet been identified. The purpose of this research is to test the hypothesis that certain kind of deformities are caused by tadpole- tadpole predatory interaction under crowded conditions. If this hypothesis is true, then tadpoles raised under crowded conditions will generate deformities while those raised under less crowded conditions will not. Faculty Supervisor: Dr. S.K. Sessions.

Possibilities for the intentional introduction of nitrogen-fixing microorganisms into agricultural environments. WILLIAM DEVOE, Department of Biology, Hartwick College, NY 13820. Despite the continued increase in the use of synthetic fertilizers, utilizing biological nitrogen fixation (BNF) to increase available nitrogen to crop systems remains an important goal of modern agricultural research. This paper explores current and past research into the intentional introduction of nitrogen fixing organisms into crop systems and offers possibilities for the application of these discoveries to agricultural practice. Major taxa of nitrogen fixing organisms are outlined, with a detailed discussion of laboratory and field conditions necessary to effectively harness their full nitrogen fixing abilities. In particular, attention is focused on promoting non-nodulating intra/extracellular nitrogen fixation in non-legume crops. Faculty Supervisors: Dr. Douglas Hamilton, Dr. Mary Allen.

An Evaluation of the Cardioprotective Effects of Treatment with the Antioxidant Resveratrol Following Transient Global Ischemia in Sprague-Dawley Rats. CHADWICK GARNER, Department of Biology, Hartwick College, Oneonta, NY 13820. Cardiac ischemia–reperfusion injury is known to result in oxidative stress and poor physiological recovery (Dernek 2004). Previous studies have demonstrated that the antioxidant resveratrol can enhance the recovery of myocardial cells and isolated perfused hearts if introduced before and during global ischemia (Siew S. et. al. 2007). However, the effects of resveratrol have not been studied in hearts reperfused with resveratrol directly following an ischemic insult, a time which is known to be more clinically feasible for treatment. This research addresses the hypothesis that reperfusion with resveratrol under these condition will enhance cardiac performance, as measured by left ventricular pressure, the rate of left ventricular pressure, heart rate and coronary flow. Four hearts were perfused with modified Krebs Henseleit buffer solution under constant pressure using the Langandorff apparatus, which simulated normal oxygen, pH, temperature and metabolic physiological condition. Heart performance before and after ischemia was measured using a pressure transducer balloon calibrated with Labscribe software program on a personal computer. The means of cardiac performance measures with and without resveratrol were compared using ANOVA with repeated measures and tentative results suggest that the presence of resveratrol following an ischemic event can increase the development of cardiac performance. This research is significant because heart disease is currently the number one cause of mortality within the United States and this experimental treatment could eventually be incorporated into clinical guidelines to help ameliorate myocardial injury resulting from ischemia. Faculty supervisor: Dr. Laura Malloy.

What's in the Milk? The Effects of Insulin-like Growth Factor I on Mouse Development. MEGHAN GEORGE, Department of Biology, Hartwick College, Oneonta, NY. Seeking to increase dairy milk production, some dairy farmers have employed a synthetic growth hormone known as bovine growth hormone (bGH) or bovine somatotropin (bST). Studies conducted by The U.S. Food and Drug Administration with the recombinant form of bST (rbST) have shown that the amount of bST in milk from injected cows is not statistically different from the amount of bST in milk from natural, non-injected cows. Thus the FDA has endorsed the use of this Monsanto product. However, the same studies have shown that the concentration of insulin-like growth factor-I (IGF-I) in milk from rbST cows is significantly increased compared to controls. IGF-I is a natural protein hormone that stimulates cell growth and tissue differentiation, while also binding to insulin receptors that regulate glucose and amino acid uptake into the cell. Even though proteins break down in the digestive tract, it is possible that they have an effect on the cells of the digestive tract before breakdown or that their breakdown products cause biological effects. Therefore I tested the hypothesis that increased levels of IGF-I in the diet could induce insulin resistance in mice. A group of juvenile mice were randomly split into two treatment groups and a control group. The two treatment groups were orally fed either 0.1ng/ul x 26ul or 1ng/ul x 26ul for 19 days. Pre and post treatment glucose tolerance tests, and percent body fat were compared to each other and to the control group. Preliminary data analysis using ANOVA suggests that the mice in the high dose treatment group showed developing signs of insulin resistance. However there was no major change in the body fat percentages across the test groups. I conclude that increased levels of IGF-I may cause metabolic changes in the body which could potentially lead to serious diseases such as Type 2 diabetes mellitus. Faculty Supervisor: Dr. Laura Malloy.
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Modeling Mildew Maturation and Detailing Dogwood Diseases. ANEES GHARZITA. Department of Math, Hartwick College, Oneonta, NY. I completed the first step in developing a mathematical model that can be applied on campus. I studied a model that predicts the timing of infection of grapevines in Italy by a mildew in hopes that I could alter it to model the life cycle of a fungus that is killing a population of Flowering Dogwood trees behind the Johnstone Science Center. The infection cycle was broken down into several stages; each stage has a “switch” that predicts the continuation of the cycle to the next stage depending on environmental conditions such as humidity and temperature. Over the course of this project, I hoped to do three things: learn about the construction of different types of math models with biological applications, understand how to develop one myself and apply it to an area of biology I wanted to learn more about. This project was undertaken as a part of a directed study called Math Modeling in Biology with Dr. Brzenk.

Genetic variation of a small isolated population of flowering dogwood (Cornus florida L.) in Otsego County, NY. ANEES GHARZITA, Department of Biology, Hartwick College, Oneonta, NY 13820. The population of flowering dogwood on the Hartwick College campus is both small (=100 individuals) and isolated. Small populations are likely to have low genetic diversity as a result of inbreeding and genetic drift. I am comparing compare the genetic diversity of a small isolated population of flowering dogwood (Cornus florida) on the Hartwick College campus in Oneonta, NY to that of a random sample of individuals from a larger nearby population. To this end, I have been extracting genomic DNA from at least 30 individuals in each population, amplifying with PCR microsatellite loci with known polymorphisms and run agarose electrophoresis gels. I will measure genetic variation as the proportion of loci that are polymorphic, the number of alleles per locus and the heterozygosity of each population. If I know how genetically at risk this population is, I may be able to suggest ways of improving or maintaining its genetic viability. Faculty supervisors: Dr. Peter Fauth, Dr. Douglas Dorer

The Correlation between Vitamin A Deficiency, Protein, and Zinc Deficiency in Malnourished Children Ages 0-5 in the Hill Tribe Villages of Northern Thailand. JOSEPH HALL, Department of Biology, Hartwick College, Oneonta, NY 13820. The hill tribes of northern Thailand have shifted from a swidden to subsistence agriculture, resulting in limited land and poor soil. As a result, this has been detrimental on the nutrition and health of the individuals, and especially the children. Malnourishment is defined as inadequate intake of macro- and micronutrients and often results in a condition known as growth stunting in children ages 0-5. This study measured serum retinol binding protein, albumin, and zinc levels to determine if the availability of these nutrients correlated with growth stunting as determined through height-age Z-scores. Anthropometric data were obtained to determine height/age Z-scores (ANTHRO program from W.H.O.). Blood samples were taken through finger prick and analyzed to determine serum nutrient levels. The correlation calculations indicated that there were significant correlations between protein and zinc (p<.002), zinc and height/age Z-scores (p<.000), protein and the height/age Z-scores (p<.049), and zinc and albumin levels (p<.000). These data indicate that both zinc and protein deficiencies play a role in growth stunting. As zinc plays a role in growth of both the plant and the consumer, analysis of zinc availability in the soil is an important factor to consider in future studies. Faculty Supervisor: Dr. Linda A. Swift

Sub-strains of Mycobacterium avium subspecies paratuberculosis strain C: Incidence and Frequency in a Central New York Holstein herd. JANETTE MARSHFIELD, Department of Biology, Hartwick College, Oneonta, NY 13820. Mycobacterium avium subspecies paratuberculosis (MAP) is an obligate parasite which causes granulomatous enteritis in ruminants. This resilient, highly contagious bacterium infects animals through the fecal oral route. With no cures for infection, this terminal disease can debilitate production in animal agriculture. In this study strain C, the strain of MAP which infects cattle, was evaluated in four Holstein cows that tested positive for MAP when tested by an independent lab: Cornell University’s Animal Health Diagnostic Center. Primers specific for the subspecies paratuberculosis were used to screen for the presence of MAP in DNA extracted from fecal samples of these cattle. Sub-strains of strain C MAP have been identified in other studies and using MAP DNA from the animals in this study the incidence of sub-strains was also investigated. Single sub-strains may be herd specific or multiple sub-strains may circulate within a herd and it could be possible for an individual animal to be infected with more than one sub-strain. Faculty supervisor: Dr. Mary Allen

The effect of dysfunctional mitochondria on the pathogenesis of Type 2 diabetes mellitus. JAMES MARTIN, Department of Biology, Hartwick College, Oneonta, NY 13820. Today diabetes affects 230 million people worldwide, 90% of them suffering from type 2 diabetes. The exact cause of type 2 diabetes (T2DM) is not fully understood, but the development of the disease relies on a genetic predisposition coupled with environmental factors. In this review the role of dysfunctional mitochondria in T2DM will be evaluated. The mitochondrial oxidative phosphorylation process in the pancreatic β-cells is integral to insulin production and secretions. Furthermore, several point mutations in the mitochondrial genome are directly associated with T2DM as well as with other accompanying diseases. Insulin resistance, one of the main characteristics of T2DM has also been associated with low mtDNA content. Mitochondria are inherited solely through the mother and a number of studies have shown that patients with T2DM have a higher frequency of mothers with this disease. This means that there is a possibility that there may be an excess maternal transmission of T2DM because of maternal inheritance of mitochondria. After reviewing all of these mechanisms current and future treatments for T2DM and mitochondrial dysfunction are evaluated in order to take another step forward into the prevention and treatment of type 2 diabetes mellitus. Faculty Supervisor: Dr. Laura Malloy

Female mate choice in domesticated guppies, Poecilia reticulata. KELLY MCGOVERN, Department of Biology, Hartwick College, Oneonta, NY 13820. Guppies have been used as a model organism for sexual selection for many years. Female guppies use phenotypic cues to choose their mates and many studies show that the female will choose the male with the highest orange coloration. These studies have focused on native guppy populations. However, the purpose of this study is to determine whether or not domesticated females follow the same mate choice trends as native guppies. A Chi-squared goodness-of-fit test will be used to evaluate the results. If there is a significant difference then mate choice may be working with artificial selection to contribute to the large variation in domesticated guppies. Faculty Supervisor: Dr. Peter Fauth

Analysis of the Correlation Between Age and Body Size in Red-Spotted Newts, Notophthalmus viridescens. APRIL NESS, Department of Biology, Hartwick College, Oneonta, NY. The purpose of this study is to examine the relationship between bone growth and body size in juvenile Red spotted newts (red efts, Notophthalmus viridescens). Newts were collected from the forest surrounding Pine Lake. Each newt was anesthetized so that the snout vent length could be measured and a single finger could be taken for sampling. The longest finger on either front limb was chosen. The fingers were then sectioned and stained using standard histological techniques. The age of each newt was estimated by counting growth rings in the cross-sectioned finger bones, and the correlation with body size was analyzed using linear regression. Faculty Supervisor: Stanley K. Sessions

Localization of the expression of Zm13 in pollen grains of Arabidopsis thaliana using the Green Fluorescent Protein. BLESSING OKOH, Department of Biology, Hartwick College. Zm13 is one of the best-studied pollen specific genes, which is expressed in the late stages of microspore development in maize, (Zea mays). It is homologous to genes that are expressed in other plant pollens that are often identified as pollen allergens. The Green Florescent Protein (GFP), which serves as a “reporter gene” will be used to tag the Zm13 protein expression. In this part of the project the Zm13 promoter region and Zm13 coding region has been linked with the Green Florescent Protein coding region, by “in frame” ligation. Polymerase Chain Reaction (PCR) primers were used to destroy the normally occurring Zm13 STOP codon which allowed the GFP to be cloned in frame, hence producing a chimeric protein. In the future this construct will be used to see if the Zm13 protein does in fact localize to the pollen wall, as do most pollen allergens. Faculty Supervisor: Dr. Hamilton.

Indirect effects between smallmouth bass, Micropterus dolemieui, rusty crayfish, Orconectes rusticus, and mottled sculpin, Cottus bairdi. RYAN OLIVER. Department of Biology, Hartwick College, Oneonta, NY 13820. Indirect effects occur when one species alters the effect that another species has on a third and are considered to be an important part of the structure of ecological communities. This experiment examines the indirect effects between rusty crayfish and mottled sculpin, which use a common refuge and are both preyed on by smallmouth bass. Trial pools with limited shelter areas were set up to observe the behavior and measure mortality rates of sculpin and crayfish with and with both the competitor and a potential predator, the smallmouth bass. The rusty crayfish is currently invading streams and lakes of northeastern North America; indirect effects such as this could play a significant role in the impact of the invasion on native communities. This study showed that sculpin do not seem to be effected in this way. No statistically significant changes in mortality or behavior can be seen in crayfish or sculpin, regardless if in the presence of bass. Faculty supervisor: Dr. Mark Kuhlmann

Localization of Osi 7 Expression By In-Situ Hybridization in Drosophila melanogaster. JON PACZKOWSKI, Department of Biology, Hartwick College, Oneonta, NY 13820. The objective of this study is to determine the expression of the Osiris 7 (Osi 7) gene in the fruit fly, Drosophila melanogaster. Osi 7 is located within the most dosage-sensitive region in the Drosophila genome, a uniquely haplo- and triplolethal locus called Triplolethal (Tpl). Dorer et al. (2003) found a family of twenty clustered genes located at Tpl which they named the Osiris family. The Osi 7 gene has been shown by RNA interference to be an essential gene in Drosophila. Osi 7 has also been found in more insect species than any of its homologs, making it the focus of this project. Using labeled DNA, fruit fly embryos will be probed at various stages of development in order to understand Osi 7’s role in the development of fruit flies. Results pending. Faculty supervisor: Dr. Douglas Dorer.
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Hypothermic Temperature Aids in the Recovery of Post-Ischemic Myocardial Tissue in Sprague-Dawley Rats. TRISTAN PETRIE, Department of Biology, Hartwick College, Oneonta, NY 13820. Coronary Heart Disease is the leading cause of mortality within the United States. Recent evidence indicates that hypothermic temperatures may assist in the recovery of the myocardium after an ischemic event, such as a heart attack. This research evaluates the effect of hypothermic temperatures (32* C) as a treatment following a brief ischemic period compared to post-ischemic hearts maintained at body temperature (37* C). Using the Langendorff apparatus, excised Sprague-Dawley rat hearts were perfused using a Krebs-Henseliet Buffer (KHB) solution that simulates physiological conditions with respect to glucose, electrolytes, pH and oxygen. Myocardial performance (heart rate, left ventricular developed pressure, rate of left ventricular developed pressure and coronary flow) were measured using a balloon pressure transducer inserted into the left ventricle, and recorded using a laptop computer. I anticipate that statistical analysis using ANOVA with repeated measures will demonstrate that decreasing temperature to 32* C for 45 minutes following ischemia results in a decline of initial myocardial function but a subsequent increase to normothermic perfusion at 37* C, will restore cold treated hearts to heart rates and cardiac performance superior to that of the control group in the same time frame. This research is significant because hypothermia could be used in the clinical setting to attenuate myocardial damage after an ischemic period. Faculty Supervisor: Dr. Laura Malloy

Molecular Cytogenetics of the Y chromosome of the North American Salamander Necturus maculosus, RYAN QUARLES, Biology Department, Hartwick College. The North American salamander genus Necturus has a unique XY sex determination system not found in closely related taxa. In this study, microdissection was used to try to isolate and amplify Y-chromosome-specific sequences in the species N. maculosus, and fluorescent in situ hybridization (FISH) was performed on meiotic and mitotic preps. A telomere probe was also used to investigate an unusual meiotic association of the N. maculosus X and Y chromosomes at their telomeres. The FISH protocol was eventually successful for N. maculosus mitotic spreads but not for meiotic spreads. It was not possible to isolate a Y-chromosome specific probe, but molecular cytogenetic studies of salamander chromosomes is very rare, and this study represents the first successful attempt to use molecular cytogenetics to study the chromosomes of this species. Faculty Supervisor: Dr. Stanley Sessions.

The Effects of Low-Fat Chocolate Milk on Exercise Performance. CHRISTOPHER SARDON, Department of Biology, Hartwick College, Oneonta, NY 13820. Previous research has shown that low-fat chocolate milk is an effective post-exercise recovery drink. It is believed that this drink has the ideal ratio of carbohydrates and protein to replenish muscles with glycogen. However, the chocolate may have other physiological effects. It can trigger the release of serotonin which acts to relieve feelings of pain and stress. The purpose of this study is to examine the mechanisms by which low-fat chocolate milk can affect exercise performance. I hypothesize that chocolate can increase time to exhaustion and total work as a result of its serotonin mediated increase in pain tolerance. 7 male participants rode a bike until exhaustion. Immediately after and 2 hours after exercise they drank 20 fluid ounces of a recovery drink (water, milk, milk and sugar, milk and chocolate, or milk with sugar and chocolate). After 4 to 5 hours of recovery, they rode again until exhaustion. Time to exhaustion, distance biked, biking speed, and perceived exhaustion, thirst and pain were compared across drinks and between the two bike rides using ANOVA. Of these measures only perceived exhaustion was significantly different between treatments, suggesting that central nervous system effects may be significant in the overall effects of sports drinks. Faculty supervisor: Dr. Laura Malloy

Behavioral Syndromes in Crayfish: Agonistic Behavior and Mating. OLIVIA VERAS, Department of Biology, Hartwick College, Oneonta, NY 13820. Behavioral Syndromes are behavioral patterns displayed consistently by individual organisms within given behavioral contexts or across several different contexts. Such behavioral patterns can lend themselves to the identification of discrete ‘personalities’ or behavioral types. As an application of the behavioral syndrome model, this study focuses on finding a correlation between increased aggression in same-sex territorial interactions and the use of agonistic mating strategies in male-female interactions. I observed 24 pairs of crayfish interact within a tank containing an artificial shelter, taking note of all aggressive behaviors displayed. 12 pairs of male and female crayfish were then observed under similar conditions. Aggression was variable within gender-specific groups and was significantly lower in female-pair interactions, but aggression levels did not significantly affect whether male crayfish employed agonistic mating strategies. This suggests that use of the agonistic mating strategy is not the extension of an aggressive behavioral pattern but derives from alternate factors; it may be a selected response to female aggression or the limited success of non-agonistic mating attempts. Faculty Supervisor: Mark Kuhlmann.
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Analysis of Extreme Heart Regeneration in the Red-Spotted Newt, Notophthalmus viridescens. SARAH WALLACE, Department of Biology, Hartwick College, Oneonta, NY 13820. A major goal of biomedicine is to understand how to induce repair and regeneration of cardiac tissue. The Red Spotted Newt, Notophtalmus viridescens, is known to be able to regenerate it limbs and to undergo repair of moderate heart damage. In this study, cardiac regeneration was analyzed by surgically removing large portions of the heart from adult newts. In most cases, newts recovered from the surgery within 24 hours. Histological analysis reveals that these newts are able to completely regenerate their hearts after removal of at least 50% of the ventricle within five weeks of amputation. These results indicate that these newts possess the ability for true, epimorphic regeneration of their ventricles. Faculty supervisor: Dr. Stanley Sessions

Effects of Short-Term Thought Triggers on Improving Student Performance in High School Biology Classrooms. WILLIAM F. WRIGHT-MCDONALD, Department of Biology, Hartwick College, Oneonta NY 13820. Educators constantly seek practices that improve learning experiences of their students. One practice, “thought-triggers”, is the use of short exercises that encourage students’ thinking prior to a class. I determined if “thought-triggers” significantly improved short-term and long-term student performance in a high school classroom. In the short-term study, I determined if a 15-minute review of concepts improved performance after a 4-week ecology unit. In the long-term study, I investigated if daily riddles used as a warm-up activity during a 1-week unit on cells improved performance. I found no significant improvement in performance during the short-term study; a 15-minute review just before taking a post-test did not improves the scores on the exam. The results of the long-term study involving riddles will also be discussed. Using a survey, I evaluated the students’ perceptions of these “thought-trigger” exercises. A review of primary literature will help to examine what other possible alternatives are available for biology educators. Faculty Supervisor: Dr. Peter Fauth.