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New Jersey Academy of Science |
Senior Academy Abstracts 2004 |
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ORAL PRESENTATIONS BIOCHEMISTRY PURIFICATION OF ?-NITROBENZOATE REDUCTASE OF RALSTONIA PICKETTII Elyse Engelhardt (student) and Dr. Theodore Chase Jr., Biochemistry Microbiology Department, Rutgers, The State University of New Jersey, New Brunswick, NJ 08901. A gene encoding a ?-nitrobenzoate reductase of Ralstonia (then Pseudomonas) pickettii, YH105 was previously cloned (Yabannavar and Zylstra, 1995). The gene was transferred to an expression vector and the protein expressed E.Coli. The substrate specificity of the enzyme was surveyed in crude extract (Campor, Zylstra and Chase, NJAS Annual Meeting, 2001). We here report protein purification of the enzyme and the initial concentration of the enzyme. The E.Coli cells that expressed the enzyme were induced with IPTG. Crude extracts were precipitated with ammonium sulfate; the precipitated enzyme was purified by gel filtration (Sepharose CL-6B) ion exchange chromatography (DEAE-Sepharose), and gel filtration (BioGel P-100). The enzyme is less stable than other nitroaromatic reductases under study in this laboratory, particularly to ammonium sulfate precipitation. A revised purification procedure is in process.
CHANGES IN GENE EXPRESSION AT THE PRECURSOR à EMBRYONIC STEM CELL TRANSITION IN LEECH Kristi A. Hohenstein (student) and Daniel H. Shain. Biology Department, The glossiphoniid leech, Theromyzon rude, displays particularly large and accessible embryonic stem cells during early development. Among these are the bilaterally-paired mesodermal (M) and neuroectodermal (N) embryonic stem cells that give rise to the bulk of segmental tissue in an adult leech, and their respective progenitor cells, DM and NOPQ. In an effort to identify genes associated with stem cell formation, pure populations of M, N, DM and NOPQ, respectively, were dissected from developing embryos. Differential display analyses were then conducted simultaneously with cDNA from all cell types. To date, ~10,000 cDNA fragments have been displayed using >100 primer sets. Approximately 200 cDNA fragments were differentially-expressed and fell into the following categories: mesoderm-specific (DM & M; 55); neuroectoderm-specific (NOPQ & N; 49); precursor-specific (DM & NOPQ; 30); stem cell-specific (M & N; 26); M-specific (15); and N-specific (39). Based on sequence analyses and GenBank BLAST searches, ~50% of these cDNAs represent novel or uncharacterized genes. Transient abrogation of several stem-cell genes with antisense oligonucleotides disrupted the normal formation of embryonic stem cells and their progeny.
PEG DERIVATIVE USED FOR BIOCONJUGATION IN ONE- AND TWO-ENZYME SYSTEMS Yuegao Huang (student), Alberto Clavijo (student), Mihaela D. Leonida, Fairleigh Dickinson University, Teaneck, NJ Polyethylene glycol-succinimidyl succinate (PEG) was used in bioconjugation procedures targeting the low stability of enzymes. Glutamate dehydrogenase (GDH) and glutaminase (Gase) were covalently linked to PEG in one-enzyme and two-enzyme settings, respectively. The activities of the pegylated systems were assessed by spectrophotometric assays and in biosensors for glutamate and glutamine detection. Following the decrease in activity due to the conjugation, the modified enzymes maintained almost constant activity for an extended time. The influence of the ratio enzyme(s):PEG on the activity of the bioconjugated systems was investigated too. The pegylated enzymes were immobilized on carbon paste electrodes together with a mediator. Different mediators and different immobilization methods were studied. The nicotinamide cofactor required for GDH was added either to the paste or in solution and the results are discussed comparatively in the paper. All biosensors studied showed pronounced catalytic effect and linearity which recommends them for use in the analysis of glutamate and glutamine in biotechnology applications and clinical lab. REGULATION OF EXTRACELLULAR MATRIX PROTEINS AND REACTIVE OXYGEN/NITROGEN SPECIES BY ULTRAVIOLET RADIATION AND TRANSFORMING GROWTH FACTOR BETA Scott
Johnson (student),
Neena Philips, Biology and Chemistry/Biochemistry Departments, Ultraviolet radiation B (UVB) damages the extracellular matrix (ECM) through an increase in the expression of matrix metalloproteinases (MMP), MMP-1 (degrades structural collagen) and MMP-2 (removes damaged collagen). Transforming growth factor-beta (TGF-b) also has effects on the ECM, specifically that it facilitates ECM deposition through the inhibition of MMP-1 and by stimulating MMP-2. Both UVB and TGF-b have been reported to independently stimulate reactive oxygen species (ROS) production and inhibit nitric oxide (NO) synthesis. We investigated the potential combinatorial influence of these two factors on dermal fibroblasts. Specifically, the purpose of this study is to investigate potential mechanisms underlying antagonistic or synergistic effects of UV and TGF-b on the expression of MMPs (MMP-1 at protein and transcriptional levels; MMP-2 at protein levels), and ROS/ NO levels in dermal fibroblasts. With the exception of a significant inhibition of MMP-1 protein level by UVB radiation, UVB radiation and TGF-b, did not alter the expression of MMPs or the reactive oxygen, and nitrogen species. UVB radiation and TGF-b in combination had beneficial potentiating effects, improved membrane integrity, inhibition of MMP-1 promoter activity, and reduced extracellular hydrogen peroxide levels, contrary to expected and may be the result of in-vitro cell passaging.
Mechanisms Regulating Elastin Expression by Ultraviolet Radiation in Keratinocytes Theresa A Lorenson (student), Neena Philips PhD., Department of Biology and Chemistry, Georgian Court University, Lakewood, NJ 08701-2697 Ultraviolet radiation has three components: UVA, UVB, UVC. UVA (315 – 400nm) and UVB (280-315nm) are found in natural sunlight. UVC (185 – 200nm) utilized in tissue culture hoods for their bactericidal affects. Excessive ultraviolet radiation exposure to unprotected skin results in upregulation of matrix-metalloproteinases’ (MMPs) transcription and tropoelastin mRNA expression, causing a decrease in collagen production along with augmenting the construction of disorganized, nonfunctional elastin. Elastin, produced by dermal fibroblasts and keratinocytes, is a fibrous component in the dermal connective tissue and is responsible for skin elasticity. In solar elastosis, elastin replaces abnormal collagen in the upper dermis. Physical symptoms of elastosis include wrinkling or sagging of the skin, poor wound healing, and leathery, thickening appearance to the skin. The goal of this endeavor is to study the affects of ultraviolet radiation on elastin manufactured by keratinocytes. Keratinocytes were transfected with elastin promoter-luciferase plasmid. Cells were either irradiated with UVA (0.06, 0.6, 1.8, or 18 J), UVB (0.25, 2.5, 7.5, or 75 mJ), or UVC (0.043, 0.43, 1.29, or 12.9 mJ) or non-irradiated. Cells and the media were harvested 8h post-UV exposure and tested for membrane integrity using LDH Assay as well as elastin production via indirect ELISA. Preliminary data indicates a compromise in membrane integrity along with indication of elastin expression by UVB and UVC in keratinocytes.
EFFECT OF SCA ON IN VITRO WOUNDING Yisel McCall (student), Samantha Vaccaro (student) Sheetal Patel (student), Lisa Karsten (student), Neena Philips, Biology and Chemistry/Biochemistry Departments, Georgian Court University, Lakewood, NJ Wound healing involves both reepithelialization and granulation tissue formation. Reepithelialization begins within hours of injury, while granulation occurs four days after incident. Both involve proliferation and migration of cells from the wound edge to fill the wound site. Once filled with the granulation tissue and covered with neoepidermis, wound fibroblast form myofibroblast. This process contracts the wound by reducing and strengthing it. Wound healing agents are continuously being identified to improve wound healing process. One such novel agent is SCA (Cantabria). Fibroblast cells were wounded using different sized pipette tips to determine which provided the best in vitro wounding for study. In addition, fibroblast cells were treated with various concentrations of SCA to assess the effect it has on the cell proliferation and membrane integrity. SCA significantly inhibited the expression of metrixmetalloprotienases and elastin, and may therby prevent improper wound healing. Experiments are being done to determine the combined effect of SCA and wounding on fibroblast cells.
INHIBITION OF FIBROBLAST CELL GROWTH AND EXPRESSION OF MMPS BY CURCUMIN AND CURCUMIN ANOLOG Yisel McCall (student), Samantha Vaccaro (student), Katherine Pytko (student) and Jennifer Garbarine(student), Neena Philips, Biology and Chemistry/Biochemistry Deparments, Georgian Court University, Lakewood, NJ Curcumin is an extract from the plant Curcumin Longa. Curcumin exhibits anti-inflamimatory and anti-infectious activities with low toxicity. It has been reported that curumin inhibits cancer and skin aging. There is an increased production of matrix metalloproteinases (MMPs), that degrade the extra cellular matrix in skin aging and cancer. In skin aging, the fibroblast are the predominant cells that produce MMPs. In this experiment the fibroblast cells were seeded and exposed to Curcumin and a water soluble form of Curcumin (Curcumin Analog). Then MTS assay was used to account for cell viability and ELISA for MMP expression. The results showed that curcumin inhibited cell growth and expression of MMP-1 and MMP-2 to a greater extent than the curcumin. These experiments are on-going and will be extended to melanoma cells.
ENERGETIC DISTINCTIONS OF ORGANISMS INHABITING TEMPERATURE EXTREMES Michael J. Napolitano (student) and Daniel H. Shain, Biology Department, Rutgers University, Camden, NJ 08102 The glacier ice worm, Mesenchytraeus solifugus, is the only known annelid that completes its life cycle in hydrated glacier ice (i.e., constant 0°C). Ice worms freeze at –6°C and begin to autolyse at 5°C. Despite their atypical habitat, ice worm behavior (e.g., motor, neural) is comparable to that of temperate earthworms. Ice organisms (psychrophiles) co-existing with the ice worm (i.e., bacteria, algae and fungi species) also survive exclusively in glacier ice. We show here that energy levels (e.g., total adenylates and energy charge) in these cold-adapted species increase as temperatures fall (even below 0°C), in direct contrast with that observed in temperate and thermophilic species. These results suggest that elevated ATP levels drive biochemical reactions in psychrophilic organisms, thus off-setting the inherent lethargy associated with cold temperature physiology.
THE EFFECT OF GLUCOSE ON THE EXPRESSION OF MATRIXMETALLOPROTEINASES IN HUMAN DERMAL FIBROBLASTS Adrienne Parratt (student) Neena Philips PhD., Biology, Chemistry/Biochemistry Departments, Georgian Court University, Lakewood, NJ Diabetes is a disease state identified by the inability to properly store or utilize serum glucose. The main characteristic of diabetes is hyperglycemia. It has been demonstrated that hyperglycemia is a principle cause of oxidative stress and a pro-inflammatory response. Long term complications of uncontrolled hyperglycemia include, blindness, kidney failure, cardiovascular disease, nerve damage, and wound healing impairment resulting in limb amputation. The mechanisms through which these deleterious effects occur are yet to be elucidated. Matrixmetalloproteinases (MMP's) are proteolytic enzymes or endopeptidases responsible for the degradation and remodeling of the extra cellular matrix (ECM) and the basement membranes of connective tissue. Altered MMP expression is observed in the wounds and systemically in diabetic patients. The present study investigates human dermal fibroblasts incubated with elevated levels of serum glucose and its effect on the discordant expression of MMP-1 and MMP-2 with its molecular inhibitors Tissue Inhibitor Metalloproteinase (TIMP), TIMP-1 and TIMP-2. No alteration in cell viability and proliferation is observed. Expression of protein and active protein will be investigated upon direct and indirect ELISA, and transcription regulation upon promoter activity will be identified using a chloramphenicol acetyl transferase (CAT) promoter.
PYROTECHNIC HEAVY METAL: EXTRACELLULAR MATRIX REMODELING BY COPPER IN SKIN FIBROBLASTS AND KERATINOCYTES Brian C Surgent
(student), Neena Philips PhD., Department of Biology and Chemistry, Georgian Court
University, Lakewood, NJ The colored visual effects of fireworks/pyrotechnics are due to the combustion of their heavy metal components. One of the heavy metal pollutants identified is copper. Copper, a widespread contaminant and an EPA priority metal pollutant, has been implicated as an essential physiological trace element and a co-factor for angiogenesis and extracellular remodeling. Our goal was to test the effect of copper on the gene expression of MMP-1, TIMP-1 and IL-8 in an in vitro system of two representative skin cell types; keratinocytes and fibroblasts. Cells were treated with increasing concentrations of the CuII ion (as copper chloride, 0, 1, 30, 300 and 1,000uM) and examined for cell viability, membrane integrity and expression of MMPs and TIMPs, mRNA expression levels and promoter activity. A significant alteration in the expression of MMP-1, TIMP-1 and IL-8 was observed, but more experiments are needed.
TUBULIN ASSEMBLY IN COLD-ADAPTED ORGANISMS Lawrence J. Tartaglia (student) and Daniel H. Shain, Biology Department, Rutgers University, Camden, NJ 08102 The ice worm, Mesenchytraeus solifugus, is the only known annelid that survives solely in glacier ice and snow. The molecular mechanism(s) of this adaptation remain unclear. Nevertheless, it seems likely that ice worms have modified their microtubules structurally, allowing them to remain functional at low physiological temperatures. Microtubules are involved in several basic cellular processes including the segregation of genetic material, intracellular transport and maintenance of cell shape. The main constituent of microtubules is tubulin, a globular protein comprising alpha- and beta- subunits that is known to depolymerize at temperatures below 10°C in most animals. In an effort to identify tubulin modifications in ice worms, both tubulin subunits were isolated via degenerate PCR (polymerase chain reaction) from an ice worm cDNA library. Following DNA sequencing, each subunit displayed several modifications in its predicted amino acid sequence, respectively. Studies of tubulin in the Antarctic fish, Notothenia corriceps suggest that there is an increased reliance on hydrophobic interactions that underlies the energetics of microtubule formation at low temperatures. An amino acid comparison between ice worm and N. corriceps tubulin subunits suggest that these two disparate animals have resolved the problem of tubulin polymerization by different strategies.
TRANSCRIPTIONAL ANALYSIS OF ELASTIN AND IL-8 IN KERATINOCYTES IN RESPONSE TO IN-VITRO WOUNDING SCA [A NOVEL WOUND HEALING DRUG], CURCUMIN AND CURCUMIN ANALOG Samantha Vaccaro (student), Yisele McCall (student), Neena Philips, Biology and Chemistry/Biochemistry Departments, Georgian Court University, Lakewood, NJ Keratinocytes form the upper layer of the skin. These cells play a critical role in wound healing and cancer via the alterations in the extracellular matrix proteins, such as Elastin and cytokines, such as IL-8. The purpose of this research was to examine the hypothesis that in-vitro wounding, SCA, Curcumin and a Curcumin analog will alter the expression of Elastin and IL-8 in Keratinocytes. Keratinocytes were seeded and transfection was done to introduce the promoter to the cells. One group of cells received the Elastin promoter plasmid while the other group received the IL-8 promoter plasmid. The cells were exposed to either SCA, wound with SCA, Curcumin or Curcumin complex, so that each promoter was exposed to a treatment. These additives did not alter cell proliferation or membrane integrity, but altered the promoter activity of Elastin and IL-8. Experiments are in progress to identify the involved mechanism.
MICROWAVE-ENHANCED SYNTHESES FOR THE UNDERGRADUATE ORGANIC CHEMISTRY LAB Danielle Falcone
(student), Dr. Marsha R. Baar, Chemistry Department, Microwave energy can greatly increase the rate of chemical reactions in which the solvent and/or a reagent has a dipole moment. The use of microwave energy as a substitute for conventional reflux heating has been investigated for a Diels-Alder cycloaddition and a Williamson ether synthesis. The Diels-Alder reaction of N-phenylmaleimide and 1,3-cyclohexadiene to form endo-N-phenyl-bicyclo[2.2.2]-oct-5-ene-2R,3S-dicarboximide requires 2.5 hours of reflux, but only 10 minutes of microwave heating to achieve a quantitative yield. The SN2 promoted Williamson ether synthesis which utilizes potassium 2-naphtholate and ethyl iodide to form ethyl 2-naphtholate (nerolin) gave comparable results with 10 minutes of microwave heating as compared to 1.5 hours of reflux. The significant rate enhancement afforded by microwave heating permits the completion of these two experiments, through purification and analysis, within one lab period.
THE ALDOL REACTION AS AN EDUCATIONAL TOOL: KNOEVENAGEL-DOEBNER REACTION REVISITED Arun Mandadi, George Barasch Bioorganic Research Laboratory, Department of Chemistry and Chemical Biology, Stevens Institute of Technology, Hoboken NJ 07030 The reaction of aromatic aldehydes with malonic esters in the presence of amines to form unsaturated esters is the century-old Knoevenagel reaction. The condensation of aldehydes with malonic acid in the presence of pyridine to form unsaturated acids is the Doebner modification of this reaction. We have found that under microwave irradiation these reactions can be completed in a few minutes. The use of the non-hazardous tertiary base TDA-1(Tris [2-(2-methoxy-ethoxy) ethyl] amine) in place of pyridine allows the reaction to be conducted in two steps. Thus, the reaction of furfuraldehyde with malonic acid in the presence of TDA-1 using 30% power level resulted in the formation of an unsubstituted dicarboxylic acid (Knoevenagel reaction step). When the reaction was performed using a high power level of microwaves, decarboxylation occurred simultaneously and an unsaturated monocarboxylic acid was obtained in good yield (as in the Doebner reaction). The synthesis of a number of mono and dicarboxylic acids using malonic acid or cyanoacetic acid and different aldehydes will be described.
LARGE SCALE GRINDSTONE CHEMISTRY FOR PROCESS DEVELOPMENT Vasudha Nekkanti (student), George
Barasch Bioorganic Research Laboratory, Department of Chemistry and Chemical Biology,
Stevens Institute of Technology, Small scale reaction between solid reactants by grinding in a mortar and pestle has been described by Prof. F. Toda and his co-workers. We were interested in a large-scale synthesis of a Schiff base that involved nitrobenzaldehyde (solid) and aniline (liquid). After some initial testing we conducted this reaction on a 50 gm scale. An electric mixture used in the kitchen was found to be a convenient equipment for large scale grinding operation. The voltage was controlled to ensure efficient mixing at a convenient rate. In less than 10 min., the reaction mixture first became a reddish liquid that solidified in a few minutes. On recrystallization from isopropanol, the target compound (Schiff base) was obtained in 94% yield.
Novel Grindstone Chemistry Mala Shah (student), George Barasch Bioorganic Research Laboratory, Department of Chemistry and Chemical Biology, Stevens Institute of Technology, Hoboken, NJ 07030 Prof. Toda of
Microwave assisted rapid synthesis of heterocycles via nitrenes Deepu Varughese (student), George Barasch Bioorganic Research Laboratory, Department of Chemistry and Chemical Biology, Stevens Institute of Technology, Hoboken, NJ 07030 The Cadogan reaction for aryl indazoles has been described in Organic Syntheses as a long process that involves a nitrene produced by triethyl phosphite at 1500C under a nitrogen atmosphere. We have used microwave enhanced chemistry to simplify this valuable method for the preparation of nitrogen heterocyles of interest to medicinal chemists. A two-step and one-pot reaction that we have developed allows all three reactants, "nitrobenzaldehyde, aniline, and excess triethyl phosphite" to be placed in an open vessel and subjected to controlled microwave irradiation. After 8 min of irradiation the target compound 2-arylindazole was obtained in 60% yield. The microwave method was used for the synthesis of different substituted indazoles as well as substituted carbazoles and indoles. These compounds were prepared on a multigram scale with high yield and purity in a few minutes of microwave irradiation. All reactions were carried out without using any solvent. We have developed an environmentally friendly method for the synthesis of aryl indazoles using water-soluble phosphines and mild microwave irradiation. Tetrakiss (hydroxymethyl) phosphonium chloride was used as deoxygenation regent instead of triethyl phosphite.
GREEN CHEMISTRY: IMPROVED SYNTHESIS OF AN ANTI-LEPROSY DRUG Olga Ziouzina (student), George Barasch
Bioorganic Research Laboratory, Department of Chemistry and Chemical Biology, Stevens
Institute of Technology, Leprosy (Hansen’s disease) is a chronic infectious disease, which usually affects the skin and peripheral nerves but has a wide range of possible clinical manifestations. We wish to report an improved synthesis of dapsone, which has been used for almost 50 years in the treatment of several infectious diseases including leprosy and malaria and recently in the treatment of infections in AIDS patients. The first step (as described in Organic Syntheses) is the reaction between 1,4-chloro-nitrobenzene and potassium xanthate in 95% ethanol and requires heating under reflux for 48 hours. Using microwave irradiation in an open system this step was complete in about 2 min; in our experiments 1,3-propanediol was used as a solvent to yield 60-65% of 4,4-dinitrophenyl sulfide. This sulfide was oxidized to the sulfone using potassium permanganate as an oxidizing agent and iron (III) chloride as a catalyst. The last step in the dapsone synthesis is the reduction of 4,4-dinitrodiphenyl sulfone to 4,4-diaminodiphenyl sulfone. Stannous chloride dehydrate/hydrochloric acid reduction of the dinitro compound under microwave irradiation resulted in the formation of dapsone in 5 min. Since tin compounds are toxic, we have developed an eco-friendly reduction step (from nitro to amino) using sodium sulfide nonahydrade under microwave irradiation for 1 min. MARINE AND COASTAL SCIENCES
INFERENCE OF TEMPORAL AND SPATIAL OCCURRENCE OF SPAWNING IN GOOSEFISH, Lophius americanus Ursula A. Howson, Department of Biology, Monmouth University, West Long Branch, NJ 07764 and R. Christopher Chambers, Howard Marine Sciences Laboratory, National Marine Fisheries Service, Sandy Hook, NJ 07732 Goosefish, Lophius americanus, is unique among the commercially valued teleosts of the Northeast USA in that females release a buoyant mucogelatinous matrix or ‘veil’ within which are numerous fertilized eggs (0.3 to 2.8 million per female). Developing embryos and newly hatched larvae are retained within this egg mass and are advected as a group from the location of spawning until the larvae hatch from the veil. Temporal and spatial occurrence of spawning is undocumented, as spawning has never been observed and egg veils are not amenable to traditional plankton sampling. Less than 30 goosefish eggs were collected in the 9000+ tows of MARMAP icthyoplankton sampling. Moreover, few studies have implemented the temporal and spatial resolution needed to determine the timing and location of spawning. Information on the timing and location of spawning is crucial for the full analysis of stock structure, understanding the role (and risk) of fish aggregations during the spawning season, and the identification of essential fish habitat for goosefish reproduction. This work sheds light on all of these features of goosefish reproductive ecology and early life history.
Use of Randomly Amplified Polymorphic DNA (RAPD) analysis as a tool for determining the genetic diversity within populations of the invasive Asiatic sand sedge, Carex kobomugi. Hendry, K. (student),
Vestyck, L, Richmyer, M and Wootton, L. Approximately 100 million
acres of land in the
IMPACT OF Carex kobomugi, ASIATIC SAND SEDGE ON NATIVE SPECIES DIVERSITY AND SPECIES RICHNESS IN COASTAL DUNES AT THE SANDY HOOK UNIT OF GATEWAY NATIONAL RECREATION AREA NEW JERSEY Myatt SL¹ (student), VanWagner C¹, Burkitt JW¹², Wootton LS¹, 1.Georgian Court University, Lakewood, NJ 2.Brookdale Community College, Lincroft, NJ In the early part of the 20th
century Carex kobomugi, Asiatic sedge, was introduced to
Impact of Carex kobomugi on plant species diversity
and species richness in VanWagner
C1 (student), Myatt S1,
Burkitt JW12, Wootton LS1. 1. Carex kobomugi
(Asiatic sand sedge) is an invasive dune species that was accidentally introduced to the
ECOLOGY EFFECT OF URBANIZATION ON WATER QUALITY OF MOLLY
ANN BROOK, Charnsmorn Hwang
(student), Michael J. Sebetich, Biology Department. Molly
Ann Brook is a small tributary to the
LIMNOLOGY OF LAC DE
LA GRISE, Edward S. Kubersky,
Department of Natural Sciences, Lac de la Grise, in
Caenorhabditis elegans: A MODEL VECTOR ORGANISM FOR ENVIRONMENTAL GENE TRANSFER STUDIES AMONG MICROBES. Luland-Richards, J.B.1(student), Adamo J.A. 1,2, Gealt M.A. 3, 1Georgian Court University, 2Ocean County College, 3Purdue University Nematodes, microscopic roundworms, occupy nearly every environment on earth. They have even been isolated in and on many invertebrates as well as vertebrates, including man. Our recent studies focused on Caenorhabditis elegans, a bacteriophagous nematode. The entire genome of C. elegans has been sequenced and its complete cellular development from zygote to adult has been elucidated. In our current studies, C. elegans was investigated as a model organism for vectoring bacteria and viruses as well as hosting horizontal gene transfers among bacteria. C. elegans was propagated in petri plates feeding on various pure cultures of bacteria. Our experimental data indicate that C. elegans is very resistant to chlorine bleach treatment, surviving exposure to 2% bleach for well over 2 minutes. This same treatment effectively controlled bacterial growth as well as viral activity. During our studies, C. elegans was found to vector viable bacteria and bacterial viruses through this treatment. This was demonstrated by surface sterilizing the nematodes and then observing bacterial growth or viral activity in selective media. C. elegans also demonstrated an ability to host the formation of transconjugants when fed alternate pure cultures of different strains of Escherichia coli, separated by surface sterilization. Bacterial strains were differentiated phenotypically by their resistance to 50 micrograms/milliliter of Kanomycin and/or 100 micrograms/milliliter of Ampicillin. USING STREAM INSECTS ON WOODY DEBRIS TO ASSESS WATER
QUALITY IN THE NEW Lynn K. Maun (student), William J. Cromartie & Julie Akers, Division of Natural Sciences and Mathematics, The Richard Stockton College of New Jersey, PO BOX 195, Pomona, NJ 08240. To resolve inconsistencies
between biomonitoring protocols and other indicators of water quality in
POSTER ABSTRACTS BIOCHEMISTRY
DEVELOPMENT OF AN INEXPENSIVE NUCLEIC ACID KINETICS EXPERIMENT FOR USE IN AN UNDERGRADUATE BIOCHEMISTRY LABORATORY Jamie Burns (student) Montclair Kimberley Academy and Amber Flynn Charlebois, Chemistry and Physics Department, William Paterson University of New Jersey, Wayne, NJ 07470 This laboratory exercise is intended to provide an opportunity for undergraduate students to be exposed to modern concepts in RNA research while using standard biochemistry and molecular biology techniques. This experiment involves studying the kinetics of a catalytic DNA molecule, a deoxyribozyme. It seems that most often nucleic acid experiments of this type involve sophisticated and/or expensive techniques that are not practical for undergraduate laboratories, including electron microscopy, and the use of radioactive or fluorescent labeling. We have developed an undergraduate laboratory that allows the student to measure the kinetics of a DNA enzyme, specifically the 10-23 DNA enzyme that was isolated by G. F. Joyce in 1998. The 10-23 is a single stranded DNA molecule that cleaves a strand of RNA site specifically. In the exercise the standard assay is completed and the products are separated using denaturing polyacrylamide gel electrophoresis and are visualized by the use of a Toluidine blue O staining process. It can be completed in a three-hour laboratory period and it exposes the student to a novel approach to enzyme kinetics.
CHEMISTRY THE ALDOL REACTION AS AN EDUCATIONAL TOOL Angelina A. Scimone, Dorina Alliu, John F. Boczany, Erica Schneider, Science Department, Caldwell College, Caldwell, NJ 07006 Although students are exposed to the concept of electron donation and electron withdrawal effects of substituents on reaction yields there are relatively few laboratory based experiments that make use of comparisons of this type. Three senior chemists performed a series of aldol reactions of methyl, methoxy, and chloro substituted acetophenones with anisaldehyde. In general, those derivatives with electron withdrawers gave higher yields that those with electron donors. In fact, these yields could be loosely linearly correlated with pKa values of the corresponding benzoic acid derivatives. Electron withdrawers should stabilize the initial enolate formed from the acetophenone moiety. Stabilization of this anion should tend increase yields especially since the anion is formed in the rate determining step. UV and IR analysis of the products confirmed conjugation. Analysis of the melting points suggested that para derivatives have a higher melting point than their ortho and meta counterparts.
MICROBIOLOGY ANTIBIOTIC
RESISTANCE IN THERMOTOLERANT FECAL INDICATOR BACTERIA ISOLATED FROM MIGRATORY Shane
Huch (Student)
and June Middleton, Biology Department, Thermotolerant
fecal indicator organisms carried by migratory waterfowl may serve as reservoirs of
antibiotic resistance. As part of an ongoing survey, we examined the fecal indicator
organism pattern of antibiotic resistance in Enterococcus
spp. and Escherichia coli isolated from the
feces of migratory
NEUROBIOLOGY Effects of Myelin Basic Protein
Mutation on Neurological Functions: A Study of Christopher
T. Suznovich (student) and Rongsun Pu, Department of Biological Sciences, Previous
studies have shown that the calcium/calmodulin-dependant protein kinase type II (
MARINE SCIENCE ASSESSMENT
OF SEABED MORPHOLOGY IN Michael J. Kennish, Scott M. Haag, Gregg P. Sakowicz, and Richard A. Tidd, Institute of Marine and Coastal Sciences, Rutgers University, New Brunswick, NJ 08901. Acoustic
records collected over a swath of estuarine floor in
ASSESSMENT OF EELGRASS (Zostera marina) RESTORATION IN Beth Wolek1, Paul Bologna1, Michael Sinnema2.
1. Department of Biology and Molecular Biology, Previous investigations into
the spatial coverage of seagrasses in
SCIENCE EDUCATION BLOOD CHARACTERISTICS OF CHICKENS (Gallus gallus) Dorfman, D. Biology
Department, A study of plasma and
hemoglobin (hgb) of adult (hen) and embryonic chicken blood was made by examination of
their electrophoretic patterns. Blood was collected from the subclavian artery in the
adult bird wing, and from the embryonic chicken at 16 through 18 days of incubation, by
piping the shell. Blood in both instances was collected in heparinized tubes, centrifuged,
and the red cells (subsequently frozen for hemolysis) and plasma separated. Agarose gel
electrophoresis was performed (Titan High Resolution Protein, Helena Laboratories,
BLOOD CHARACTERISTICS OF TWO POECILIIDAE (fishes) Dorfman, D., Plasma and hemoglobin (hgb)
patterns were determined and compared from blood, obtained from the caudal artery, of
mosquitofish, Gambusia affinis, and from guppies, Poecilia reticulata. Blood from female fishes was
obtained in heparinized microcapillary tubes, then centrifuged, and the plasma and red
blood cells separated. These were frozen until electrophoresed. Agarose gel (Titan High
Resolution Protein, Helena Laboratories,
BLOOD PATTERNS OF TWO MENIDIA SPECIES Dorfman, D. Biology
Department, Blood was collected from the
GROWTH OF GEOCHELONE SULCATA Dorfman, D., Biology
Department, The African spurred tortoise
(G. sulcata) is the largest mainland tortoise. It reaches weights of 200 pounds. It
occurs in the southern
Dorfman, D. This study examined the response of fishes, banded killifish (Fundulus diaphanus), and guppies (Poecilia reticulata) to waters supersaturated with oxygen. Fishes more typically face the problem of reduced dissolved oxygen, and respond by moving upwards in the water column, towards the surface of the water. On some occasions fishes may be exposed to supersaturated waters, for example below dams, and in aquatic plant blooms. To provide supersaturated waters a reservoir gravity fed water into a fractionating column. Oxygen was pumped into the column and the supersaturated waters entered into a one gallon tank containing the test fish, and a stir bar to distribute the water evenly through the tank. Within a two hour period the oxygen levels were raised from 5.0 mg O2/l to 31 mg O2/l. The responses of both species was initial agitation for both, and a movement to the bottom of the tank. Guppies rested quietly on the tank bottom, whereas the killifish were more agitated. Downward movement began at approximately 140% saturation. Saturation levels reached 375% at test temperatures of 25 C. No mortalities occurred. Oxygen was determined by the Winkler method. THE
OISTINS FISH Dorfman, D. The fish market at
BLOOD CHARACTERISTICS OF THREE SPECIES OF XENOPUS (frogs) Dorfman, D., and K.
E. Briley, Jr. Blood bands for three species of adult Xenopus, X. laevis, X. borealis, and X. tropicalis were determined and compared. Blood was collected from the iliac artery in heparinized capillary tubes. The blood was centrifuged and the plasma and red blood cells frozen. Plasma, and hemoglobin (hgb), obtained from the lysed red blood cells, were electrophoresed, for 24 minutes, using an agarose gel technique (Titan High Resolution Protein, Helena Laboratories, Beaumont, Texas). For plasma, X. laevis contained seven bands (ranging from six to 25% of the total proteins), X. borealis contained eight bands (range, six to 23%), and X. tropicalis contained six bands, with one of these cathodic (11% of the total proteins) and five anodic bands (range, 11 to 23% of the remaining total proteins). For hgb X.laevis and X. tropicalis had single bands, whereas X. borealis had two bands. Band I (closest to the point of application) contained 41% of the hgb, and band II the remainder. All hgb bands were anodic. Analysis of the blood of tadpoles of the three species is planned. YELLOW PERCH LENGTH COMPARISONS Dorfman, D., and T. Smith Monmouth University, The purpose of this study was to develop a program to relate the different methods of measuring the length of yellow perch (Perca flavescens). Length measurements have been reported in any one of several ways by researchers. For example, fish lengths may be reported as standard length, body length (tip of the snout to the origin of the tail), fork length, total length, and total length tail compressed. For this study the data consisted of the five measurements (mm) mentioned for each of 35 fish. No attempt was made to identify anomalous data. A least square linear equation was determined for each ordered pair of measurements. These equations were then used as the predictor equations for the Excel spreadsheet. To compare data for yellow perch fish length with other studies of the same species, the researcher inputs his data measurement into the program and has the potential for four other length measurements. The equations are available from the senior author for this species, and, from similar studies, for white perch (Morone Americana), bluegill (Lepomis macrochirus), and pumpkinseed (L. gibbosus).
HOLOCENE,
PLEISTOCENE, AND CRETACEOUS FOSSILS FROM BIG BROOK, NEW Harbour, G., and D. Dorfman One of the major drainage
zones in Marlboro, NJ, |
