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Undergraduate Research Symposium
The Auburn Montgomery School of Sciences Table of Contents
Schedule of Events . 5 Poster Session I . 6 Oral Session I . 7 Poster Session II. 8 Abstracts . 9-28 Phagocytic Activity in Bufo marinus . 10 Antibiogram of Coliform Bacteria Isolated from River Water . 11 Anuran Immunology and the Effect of Corticosterone on Basophil Proliferation . 12 Morphological and Behavioral Effects of an Androgenic Compound on the Eastern Mosquitofish, Gambusia holbrooki . 13 A Comparative Analysis of the Anuran, Bufo marinus, with Exposure to Atrazine . 14 Interactions of the Complement System of the Anuran, Bufo marinus, with Leukocytes . 15 The Effects of an Environmentally Induced Fever on Antibody Production in Bufo marinus . 16 The Effect of Vitamin E on the Complement System of Bufo marinus . 17 Genetic Transformation of the Bacterium Serratia marcescens . 18 The Effect of Glass Vacutainer Tubes on the Activated Partial Thromboplastin Time (APTT) . 19 The Effect of Interstate Highway Policy on Urban Neighborhoods: The Case of Montgmery, Alabama . 20 Knowledge of Heart Symptoms: Associations of History of Heart Attack, Sex and Education . 21 Corticosterone Suppression of Basophils During Inflammation in Bufo marinus . 22 Effect of Prednisone Treatment on LH Levels of Ovariectomized Mice . 23 Convexity and Quadrilateral in Euclidean Space . 24 The Effects of an Environmentally Induced Fever on Phagocytic Activity in Bufo marinus . 25 The Effectiveness of Activating Different Complement Pathways in Bufo marinus . 26 Isolation and identification of N-acyl-homoserine lactones (AHL) from pigment producing Serratia species . 27 The Auburn University at Montgomery
Schedule of Events
School of Sciences
9 a.m. - 9:15 a.m. Goodwyn Hall Room 109 9:15 a.m. – 10:30 a.m. Poster Session I The 2009 Undergraduate Research Symposium Goodwyn Hall Atrium – coffee and snacks It is our pleasure to welcome you to the School of Science's second annual 10:30 a.m. – Noon Undergraduate Research Symposium. We are excited to offer this professional venue Goodwyn Hall Room 109 for Auburn Montgomery's undergraduate researchers to present their research and to be recognized for their scientific investigations. Each presentation is a culmination of hard Noon – 12:30 p.m. work conducted by students and their faculty mentors. Goodwyn Hall Atrium - lunch provided On behalf of the School of Sciences, we trust you will find these research presentations 12:30 p.m. – 2 p.m. Poster Session II enlightening and inspiring.
Goodwyn Hall Atrium 2 p.m. – 2:30 p.m. Goodwyn Hall Atrium – snacks provided Awards Ceremony and Closing Remarks Dean of the School of Sciences Associate Dean of the School of Sciences Goodwyn Hall Room 109 Dr. Glen Ray
Dr. Matthew Ragland
Undergraduate Research CouncilDr. Bridgette Harper – PsychologyDr. Kalu Kalu – Public AdministrationDr. Jeanne Kuhler – Physical SciencesDr. Enoch Lee – MathematicsDr. Rich Martin – Justice and Public SafetyDr. Chelsea Ward – Biology Poster Session I
Oral Session I
1. Antibiogram of Coliform Bacteria Isolated from River Water 10:30 a.m. Phagocytic Activity in Bufo marinus Iesha L. Barnes Mentors: Benedict Okeke Matt Atchison, Kaori Knights, Ariel Halway Mentor: Chelsea Ward and LaTresha Roberson 2. Anuran Immunology and the Effect of Corticosterone on Basophil Proliferation 10:45 a.m. Interactions of the Complement System of the Anuran, Bufo marinus, Roselyn Basco, Ingibjorg Heiddal, Justin Hogan Mentor: Chelsea Ward and Vanessa House Haley Deal, Iral Patel and Jonathan Wolfe Mentor: Chelsea Ward 3. Morphological and Behavioral Effects of an Androgenic Compound on the Eastern Genetic Transformation of the Bacterium Serratia marcescens Mosquitofish, Gambusia holbrooki Mentor: Pryce L. Haddix Lena Bosheh, Amber Montemayor, Chris Grier Mentor: Robert Estes and Jeremy Browning 11:15 a.m. The Effect of Interstate Highway Policy on Urban Neighborhoods: The Case of Montgmery, Alabama 4. A Comparative Analysis of the Anuran, Bufo marinus, with Exposure to Atrazine Mentor: Pia Knigge Lindsey Brown, Jessica Thomas, Rod Weldon Mentor: Chelsea Ward and Tenesha Williams 11:30 a.m. Convexity and Quadrilateral in Euclidean Space Mentors: Robert Underwood 5. The Effects of an Environmentally Induced Fever on Antibody Production in Bufo marinus Luis Cueva-Parra Micah Dettmar, Krysten Durham, Tamara French Mentor: Chelsea Ward and Sherilyn Gitchel 11:45 a.m. Effect of Prednisone Treatment on LH levels of Ovariectomized Mice Mentor: Nathan Okia 6. Corticosterone Suppression of Basophils During Inflammation in Bufo marinus Ankit Patel, Shivam Patel, Shanticia Peaks Mentor: Chelsea Ward and Andrain Robinson Poster Session II
7. The Effect of Glass Vacutainer Tubes on the Activated Partial Thromboplastin Time (APTT) Jonathan Hendricks Mentor: Virginia C. Hughes 8. Knowledge of Heart Symptoms: Associations of History of Heart Attack, Sex and Education Brittany N. Miller and Stephanie Holman Mentor: Steven LoBello 9. The Effect of Vitamin E on the Complement System of Bufo marinus Alisha Hardy, Jamie Harris, Chelsie Johnson Mentor: Chelsea Ward and Rushi Patel 10. The Effects of an Environmentally Induced Fever on Phagocytic Activity in Bufo marinus Stephanie Smith, Brittany Stockton, Jeleria Williams Mentor: Chelsea Ward and Brittany Wright 11. The Effectiveness of Activating Different Complement Pathways in Bufo marinus Aya Zaied, Dennis Lawerence, La Carlos Wright Mentor: Chelsea Ward 12. Isolation and identification of N-acyl-homoserine lactones (AHL) from pigment producing Serratia species Dominique N. Lewis and Jonathan T. McFarland Mentor: Duk K. Kim and Phagocytic Activity in Bufo marinus
Antibiogram of Coliform Bacteria Isolated from River Water
Matt Atchison, Kaori Knights, Ariel Halway and LaTresha Roberson Mentor: Chelsea Ward Mentors: Benedict Okeke and Sue Thomson Recently amphibian populations have been declining at an alarming rate, especially Antibiotic sensitivity pattern of coliforms can be used for source tracking. Coliforms in pristine tropical areas. Decreases in immune function have been hypothesized to be such as Escherichia coli, Enterobacter sp. and Klebsiella sp. are gram-negative rod- a cause of the amphibian die-offs. However, baseline immune functions have not been shaped, non-spore forming bacteria that metabolize lactose with the production of acid quantified. We have been working to describe the immune function of the marine toad, and gas when incubated at 35-37°C. The presence of coliforms in water resources is Bufo marinus. We evaluated the circulating levels of white blood cells and how those a strong indication of human or animal waste pollution. Detection of E. coli in water levels interact with phagocytic activity. Blood samples were collected from toads bi- invariably suggests potential presence of disease-causing microorganisms, including weekly for six months. Phagocytic activity was evaluated by mixing blood cells with a viruses and parasites. known quantity of E. coli and allowing the phagocytic cells to kill the bacteria. After a In this study, coliform bacteria were isolated from river water serving a water set incubation period, the surviving E. coli were allowed to grow on an agar plate and treatment plant. Colony color on Easygel was used to screen isolates from different counted. Differential field microscopy was used to determine white blood cell quantities seasons (spring, summer and fall) to identify E. coli. Nine bacterial isolates selected from for each blood cell. The data was then correlated in an attempt to determine overall Easygel plates were identified by 16S ribosomal RNA gene sequence. Five isolates were immune function. identified as E. coli, three as Enterobacter spp. and one isolate as Klebsiella sp. Analysis of sensitivity of the isolates to gentamicin, trimethoprim/sulfamethazole, ciprofloxacin, vancomycin, tetracycline, ampicillin, cefixime and nitrofurantoin revealed a similar profile for the E. coli isolates. All the nine isolates were resistant to vancomycin. The E. coli isolates were most sensitive to ciprofloxacin (40 – 42 mm zone of inhibition) and cefixime (24 – 30 mm zone of inhibition). The antibiogram of the E. coli isolates suggests they may originate from a common source.
Anuran Immunology and the Effect of
Morphological and Behavioral Effects of an Androgenic
Corticosterone on Basophil Proliferation
Compound on the Eastern Mosquitofish, Gambusia holbrooki
Roselyne Basco, Ingibjorg Heiddal, Justin Hogan and Vanessa House Lena Bosheh, Amber Montemayor, Chris Grier and Jeremy Browning Mentor: Chelsea Ward Mentor: Robert Estes This experiment explores the effect of corticosterone on the proliferation of Contamination of the environment with endocrine disruptors, chemicals that mimic basophils. Basophils are the smallest and least common leukocytes. Although the hormones, has become a widespread problem. Studies have linked contaminants in function of basophils is unclear, recent research has shown that basophils might play a sewage wastewater and pulp mill effluent to morphological alterations in some organisms role in the response to a parasitic infection. When activated, basophils are believed to living downstream of these facilities. The mosquitofish, Gambusia sp., has been shown to discharge the contents of their granules to release chemicals like histamine, serotonin and undergo specific transformations when exposed to endocrine disruptors. In the presence cytokines, which contribute to an inflammatory response in the body. Lipopolysaccharide of androgenic contaminants, the anal fin of the female mosquitofish is masculinized and (LPS) is known to mimic inflammation involved as a result of bacterial infections in the resembles a gonopodium, an elongated anal fin normally present only in males. species Bufo marinus. Although physical alterations have been well established in this species, This experiment will use LPS to induce an inflammatory response in Bufo marinus. behavioral changes have not been sufficiently addressed. We seek to determine effects Corticosterone will then be administered to the Bufo marinus. An increase in stress of androgens on social and reproductive behaviors of female mosquitofish. We exposed hormones has been shown to have a negative effect on the immune system. As a result, wild-caught fish to the androgenic compound adrenosterone and will subsequently an increase in corticosterone level is expected to decrease the basophilic response to the determine the effects on specific behavioral patterns of female mosquitofish. Fish were LPS induced inflammation.
housed in 1-liter beakers and treated with adrenosterone for five weeks to induce anal fin transformation. Transformed fish will be subjected to observational studies to determine effects of transformation on behavioral patterns of these fish when combined with either wild type or other transformed fish. These studies will shed light on the effects of endocrine disruptors in a vertebrate model and may provide insight into potential effects on higher organisms, including humans. A Comparative Analysis of the Anuran, Bufo marinus,
Interactions of the Complement System of the Anuran,
with Exposure to Atrazine
Bufo marinus, with Leukocytes
Lindsey Brown, Jessica Thomas, Rod Weldon and Tenesha Williams Haley Deal, Iral Patel and Jonathan Wolfe Mentor: Chelsea Ward Mentor: Chelsea Ward Eosinophils are leukocytes that fight against the invasion of parasites and in many Little is known about the interactions of leukocytes with the complement system cases are prevalent during allergic responses. Considering this white blood cell as the of frogs. An increase in either the quantity of leukocytes or complement proteins can basis, the decision to take an immunosuppressant approach for experimentation was indicate an infection. This experiment compares the percentage of leukocytes in the blood determined. The question our group poses is, "What will lower the amount of leukocytes stream with the lytic activity of the complement system of the anuran, Bufo marinus. The in the blood and specifically the presence of the eosinophils?" Atrazine is the most widely lytic activity of the complement system is determined through the lysis of Escherichia coli used form of herbicide in the United States and would be very accessibly reached by resulting from its mixture with complement proteins from frog plasma. The percentage such amphibians as the experimental toads (Bufo marinus). of leukocytes in the blood is obtained through a manual complete blood count. The After an initial white blood cell count, we will present increasing, yet non-lethal, complement protein production can be increased by antibodies binding to their targets. amounts of atrazine to the experimental toad. This will be done over time through dilution Leukocyte production can be increased by complement proteins binding to pathogens. In in the drinking water just as they may encounter in the wild. Each week a blood sample addition, both systems can increase their production by themselves. The close interactions will be taken from both toads for a blood smear. The Giemsa-Wright staining technique between the complement system and leukocytes show that the percentage of leukocytes in will be used for easy identification of the leukocytes during a differential white blood cell the blood will change with the lytic activity of the complement system.
count. We hypothesize to see a decrease in eosinophils, as well as a lower overall white blood cell count in the Atrazine-exposed toad as compared to the control toad.
The Effects of an Environmentally Induced Fever on
The Effect of Vitamin E on the Complement
Antibody Production in Bufo marinus
System of Bufo marinus
Micah Dettmar, Krysten Durham, Tamara French and Sherilyn Gitchel Alisha Hardy, Jamie Harris, Chelsie Johnson and Rushi Patel Mentor: Chelsea Ward Mentor: Chelsea Ward When an endotherm immune system is responding to an immune response, it lets Previous studies have shown that vitamins, E in particular, enhance the ability off heat or a fever to aid in fighting an infection. It is not known whether an increase in of the immune system to protect the host from lethal bacterial infections. Vitamin E temperature would likewise aid an ecotherm in fighting an infection. To test this we are has been shown to enhance complement activity in chickens infected with Escherichia using the marine toad, Bufo marinus. We injected two toads with lipopolysaccharides to coli infections. We investigated the effects of vitamin E on the complement system in simulate a bacterial infection. Sheep red blood cells (a novel antigen) were also injected the anuran, Bufo marinus. One frog will be fed supplemental vitamin E at the level of into the frogs to induce antibody production. One toad was kept at 20 °C and the other 300mg/kg of body weight. We will then perform a bacteria killing assay in order to at 18°C . We will then test antibody production weekly in both toads by exposing toad determine the effectiveness of the vitamin E to enhance complement activity.
plasma to sheep red blood cells and measuring the amount of hemagglutination. We believe that the higher temperature will serve as an environmentally induced fever and aide the toad in fighting infection by increasing antibody production. Genetic Transformation of the Bacterium Serratia marcescens
The Effect of Glass Vacutainer Tubes on the
Activated Partial Thromboplastin Time (APTT)
James A. HauseMentor: Pryce L. Haddix We have developed a method for transformation of the bacterium Serratia Mentor: Virginia C. Hughes marcescens. Transformation, the addition of material to the cellular genome, requires uptake of a gene which confers a survival advantage on bacteria receiving experimenter- The Activated Partial Thromboplastin Time (APTT) is performed in the clinical added DNA. Growth of S. marcescens was inhibited with ampicillin at 500 µg/mL; this laboratory to determine if a patient has a clotting factor deficiency, circulating antibiotic concentration was used to isolate bacteria transformed with DNA encoding anticoagulant, or to monitor heparin therapy. The reagents used in the APTT include a ampicillin resistance. Our transformation procedure utilized high-voltage electrical fields contact activator, phospholipids and calcium chloride. The contact activator is typically to disrupt the bacterial cell wall and promote DNA uptake. The optimum voltage was a suspension of negatively charged particles such as kaolin or ellagic acid. Blood is found to be 2.5 kV. S. marcescens cells grown at 37°C exhibited high endogenous collected into either plastic or glass vacutainer tubes containing sodium citrate as an ampicillin resistance, which made them unsuitable for transformation with DNA encoding anticoagulant at a ratio of 9 to 1. resistance to ampicillin. However, successful transformation to ampicillin resistance was This study sought to investigate whether the negatively charged surface associated achieved, albeit at low frequency, for cells grown at 30°C.
with glass tubes significantly affected the APTT result in a paired design (n = 10) using glass and plastic tubes. Using abnormal controls, 0.5 mL citrated plasma was aliquoted into either 12 x 75 mm glass or plastic tubes with a unique identifier and refrigerated overnight. The samples were analyzed for an APTT using the Electra 750 photo-optical plasma coagulation timing instrument. The test involved adding 0.1 mL of test plasma to 0.1 mL of APTT activator (ellagic acid) and incubated at 37C for three minutes. Following incubation, 0.1 mL of calcium chloride was added and photo-optical determination of a fibrin clot was measured in seconds. Using a paired t-test, significance was found between the glass and plastic samples. (P = 0.009) This knowledge is useful in the event glass vacutainer tubes are used in the APTT test and analysis is delayed.
The Effect of Interstate Highway Policy on Urban
Knowledge of Heart Symptoms:
Neighborhoods: The Case of Montgmery, Alabama
Associations of History of Heart Attack, Sex and Education
Brittany N. Miller and Stephanie Holman Mentor: Pia Knigge Mentor: Steven LoBello The trans-American Interstate Highway system drastically altered the urban American According to the Centers for Disease Control, heart disease is the primary cause of landscape, setting the stage for the era of the populace suburb surrounding the desolate death in the United States; therefore, it is vital to know the warning signs. This study is an urban center, the diffusion of economic resources over a greater land area, and the era of analysis of the differences in knowledge of heart attack symptoms for different education white flight and modern segregation. However, the placement of the highway system, as levels and genders among individuals who have or have not experienced a heart directed by state and local governments, served as a targeted weapon to clear slums and attack. Data are from the 2007 Behavioral Risk Factor Surveillance Survey produced redevelop urban cores. In many jurisdictions this weapon was used to destroy African- by the Centers for Disease Control. The purpose of this study is to determine if previous American middle class neighborhoods. Montgomery, Alabama, presents a perfect case heart attack is associated with an increase in knowledge of symptoms. Furthermore, study on the targeted impact of the Interstate Highway System to destroy the African- because women experience different heart attack symptoms than men, we also looked American middle class and the lasting effects of such a decision. A longitudinal study of at differences in knowledge of symptoms between women who have and have not socioeconomic factors such as property value and demographic statistics where available had heart attacks. The findings indicate that women who have suffered a heart attack of several census tracts targeted by the construction versus the city as a whole will show are more aware of symptoms such as jaw pain, chest pain, arm pain and shortness of the disparate impact of this policy on the African-American population of this Southern breath, while men are more aware of weakness. The findings also indicate that high town. By studying these statistics over the decades beginning in 1960, the adverse school dropouts who have suffered a heart attack are less likely to recognize heart attack effects, both intentional and unintentional, of the placement of the Interstates.
symptoms than individuals who have suffered a heart attack but did graduate from high school, attend or graduate from college.
Corticosterone Suppression of Basophils
Effect of Prednisone Treatment on
During Inflammation in Bufo marinus
LH Levels of Ovariectomized Mice
Ankit Patel, Shivam Patel, Shanticia Peaks and Andrain Robinson Mentor: Chelsea Ward Mentor: Nathan Okia Basophils have been known to have a substantial contribution on the inflammatory Prednisone is a synthetic glucocorticoid drug. Previous works have suggested that response. We are attempting to determine the effect of corticosterone, a stress hormone, both excess and stress glucocorticoids are effective endocrine disruptors, responsible in the presence of ovalbumin on basophil production. Ovalbumin can be used as an for several reproductive or developmental complexities. Several studies have also antigen to cause an inflammatory response that causes an increase in basophils. This demonstrated that prednisone treatment inhibits luteinizing hormone (LH) secretion by happens due to basophils' key characteristic of being the major cells produced during the acting at the hypothalamic and/or pituitary level in humans and dogs. In view of this, inflammatory response. Corticosterone is a chemical that has been known to act as an we examined the relationship of prednisone treatment to the levels of LH using an ELISA anti-inflammatory reagent. Corticosterone decreases the number of basophils that can be in ICR mice. Since it is known that gonadal secretions act to inhibit GnRH secretions produced by lowering the amount of mediators the inflammation response releases such at hypothalamic and/or pituitary level via negative feedback, in the present study, we as histamine. In our experiment, two Bufo marinus are injected with ovalbumin twice will employ ovariectomized mice for enhanced response. Mice will be subcutaneously weekly for a six-week time period. During these six weeks, only one of the frogs will be injected with 0.1 mg/kg/day (n=5), 0.5 mg/kg/day (n=5) and 1.0 mg/kg/day (n=5) injected with corticosterone once a week to determine if stress hormones have an effect of prednisone for 14 days. Mice in the control group (n=5) will receive only corn oil. on basophil production even in the presence of an antigen. Blood smears will be done Following the 14-day regiment, LH levels will be measured using ELISA.
to observe the effect of the different dosages on basophil production. We propose that Bufo marinus not injected with corticosterone will produce a significantly higher number of basophils over the six-week period.
Convexity and Quadrilateral in Euclidean Space
The Effects of an Environmentally Induced Fever
on Phagocytic Activity in Bufo marinus
Pratik P. Patel
Mathematics
Stephanie Smith, Brittany Stockton, Jeleria Williams and Brittany Wright Mentors: Robert Underwood and Luis Cueva-Parra Biology
Mentor: Chelsea Ward
A quadrilateral is said to be convex if each of its sides lies in one of the half planes determined by the opposite side. The converse is trivial. Thus, ABCD is a convex Phagocytes are part of the innate immune system, ingesting microbes and other quadrilateral if and only if all four of the following conditions hold: 1.) A and B are on non-self material in the body. The purpose of the experiment will be to determine the same side of line CD; 2.) B and C are on the same side of line DA; 3.) C and D are whether or not temperature will have an effect on the optimal function of the immune on the same side of line AB; 4.) D and A are on the same side of line BC. Conjecture: If system. Bufo marinus were given a pseudo-infection by injecting them with a piece of quadrilateral ABCD fails to be a convex quadrilateral, then two of the four conditions fail a bacteria cell wall, lipopolysaccharide. Toads were then housed at two temperatures, and the other two conditions hold.
one at 20 °C and the other at 18°C. The production and activity of the phagocytes will be measured by luminol-enhanced chemilunescence stimulated by zymosam (a yeast product). As phagocytic activity increases, luminal is activated and light is given off. The amount of light produced is measured and quantified. If phagocytic activity is related to temperature, then the toad blood should have an increased phagocytic activity when given a pseudo-bacterial infection and placed in a warmer environment. The Effectiveness of Activating Different
Isolation and identification of N-acyl-homoserine lactones (AHL)
Complement Pathways in Bufo marinus
from pigment producing Serratia species
Aya Zaied, Dennis Lawerence, La Carlos Wright and Lena Bosheh Dominique N. Lewis and Jonathan T. McFarland Physical Sciences and Biology
Mentor: Chelsea Ward Mentors: Duk K. Kim and Pryce L. Haddix Compliment is composed of three distinct pathways: the classical, the alternative Serratia marcescens is an opportunistic bacterial pathogen found in water, soil, and the mannose binding lectin pathways. Although all three are activated by different milk, foods, and insects. It can cause infections of the endocardium, blood, wounds, mechanisms, all culminate with the creation of the membrane attack complex which urinary tract, and respiratory tract. Prodigiosin pigment is a three-ringed hydrophobic functions to destroy antigens. Because all three converge and conclude at the same molecule associated with the bacterial inner membrane. Many functions have been point, the question can be raised as to whether any one of the three is more effective proposed for prodigiosin, including a role as an antibiotic. Many gram-neragive bacteria or efficient than the other two. Because the classical pathway is reliant on elements of regulate the expression of specialized gene sets in response to their population density the adaptive immune system, specifically antibodies bound to the surface of an antigen, by a mechanism called autoinduction. These bacteria measure their cell densities by a conclusion might be drawn that this pathway is less efficient because activation sensing the level of a chemical signals, called the autoinducer (AI). All AIs identified cannot begin until the antigen is "tagged" by an antibody. Because the alternative up to now are N-acylated derivatives of L-homoserine lactones (L-AHLs). Our research and lectin pathways don't rely on antibody activation, it is more difficult to determine work focuses on the isolation of pure AHLs from the bacterial cultures for further study to which pathway is more effective and efficient. By up-regulating both the alternative control of bacterial growth with the purified AHLs. Preparative scale reversed phase thin and lectin pathways experimentally, it is possible to determine if either pathway is more layer chromatography (TLC) was applied for the primary isolation of AHLs. AHL spots effective at killing antigens. Using GDP-mannose to activate the lectin pathway and will be visualized with biosensor strain. An HPLC technique will be used to purify each lipopolysaccharide to activate the alternative, we can test whether one pathway is more ALH. A variety of analytical techniques of Infrared (IR) spectroscopy, Nuclear Magnetic effective at creating the membrane attack complex than the other. Resonance (NMR) spectrometry, and Mass spectrometry will be applied to identify chemical structure of the isolated AHL. A presentation will be given to explain how our works have been done and how we have analyzed the analytical data obtained from the proposed work shown above.

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