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BIOLOGY

 

2000 Eastern College Science Conference
Wagner College, Staten Island, NY


College of Mount Saint Vincent and Manhattan College faculty and students attending the 2000 Easter College Science Conference on April 1, 2000.Top row: Evangelos Pefanis, Anthony Mastropietro, Dr. Brian Fee, Dr. Michael Judge, Dr. Annie Visviki, Dr. Lance Evans. Second row: Dmitriy Zamarin, Christina Doyle, Pascale Rabbah, Jessica Papile, , Dmitra Doupis.
Third row: Dr. Bill Tramontano, Lidia Prokopowicz. Fourth Row: Marigrace Lim, Elvira Liclican, Jackie Pancrudo.
Fifth Row:
Dr. James Haley, Vina Cruz, Diane Craft, Anne Paul, Karen Lagrazon.

Awardees:
Manhattan College 2000 students


Best Biology Paper - Elvira Liclican
Best Oral in Biology - Marigrace Lim
Best Oral in Molecular Biology -
Evangelos Pefanis


Click on the author(s) to see their abstracts. Then click on the Browser back button to return to this page.


Diane Craft and Michael Judge
Vina Cruz
Dimitra Doupis, Nada Assat-Anid, and Walter W. Faber Jr
Christina A. Doyle and Brian E. Fee
Elvira Liclican
Marigrace Lim
Jacklyn Pancrudo and Karen Lugrazon
Jessica Papile, James Murphy, and William A. Tramontano
Anne Paul
Pascale Rabbah and Ioanna Visviki
Dmitny Zamarin, Kai-i Anderson, Jason Paragas, and Peter Palese

THE EFFECT OF THE TUNICATE, MOGULA MANHATTANENSIS, ON MARINE MICROBIAL GROWTH.
Diane Craft and Michael Judge. Biology Department. Manhattan College/College of Mount St. Vincent. Riverdale. NY 10471
Many marine organisms, including tunicates. possess biologically active compounds that function against microbes and other organisms. These compounds may play a vital role in survival because they are metabolically expensive, structurally complex, and found in high concentrations. The effect of the temperate subtidal tunicate. Molgala manhattanesis, on co-occurring bacteria was determined by zones of inhibition on a disk assay experiment. Water samples and tunicate specimens were collected from a marina (Bronx. NY) during fall 1999. One bacterial species was isolated and identified using various growth media and the API 20E system. The tunicates were frozen, divided into visceral and tunic components, and homogenized separately via Waring blender. Utilizing five different treatments (homogenized viscera, homogenized tunic. 3.0 (um Filtered seawater. sterile seawater. and distilled water), the disk assay experiment revealed no differences among the zones of inhibition (all means = 0 mm). Results suggest that M. manhattanesis did not exhibit any pronounced anti-microbial activity towards this species. Thus, M. manhattanesis has not developed effective defenses against all naturally co-occurring bacteria.

SELECTIVE PARTITIONING OF NEURAL CELL ADHESION MOLECULES (N-CAM) ISOFORMS WITH TRITION X-100.
Vina Cruz. Department of Biology. Manhattan College, College of Mount Saint Vincent. Riverdale. NY 10471
Astrocytes play a critical role in Neurobiology in several aspects: in the balance of K.' to sustain the neuronal environment, in brain development, in neurocransmission. and in formation of the blood-brain barrier. Astrocytes contain a cell adhesion molecule. N-CAM. which participates in cell-cell recognition and interaction. N-CAM. a cell surface glycoprotein. participates, in part, in the aforementioned brain functions of astrocytes in the CNS. During brain injury and other neurological diseases, reactive astrocytes produce glial scars, which result in gliosis and these cells disrupt CNS neurons from regenerating. This astrocytic gliosis inhibits the axon from penetrating the glial scars. Recently, N-CAMs. in culture, have shown to inhibit the proliferation of astrocytes and perhaps this may have clinical ramifications.. In this research. N-CAM. from astrocyte cultures, selectively partitioned into a Triton insoluble, cytoskeletal. fraction (Tl) and into a Triton soluble fraction (TS) were first isolated via centriftigation. Next. examination of Western blots of two-dimensional polyacrylamide gels showed that the lower molecular weight isoform N-CAM (~l20Kd) partitioned into the Tl whereas two higher molecular weight isoform N- CAMSs (-160 and 200 Kd) into the TS. Therefore, inhibition of astrocyte proliferation by N-CA'M may be associated with the cvtoskeleton for anchorase.

FALSE NEGATIVE AND FALSE POSITIVE RATES IN THE CONFIRMATION OF POSITIVE GROWTH IN THE TOTAL COLIFORM MEMBRANE FILTRATION ANALYSIS.
Dimitra Doupis, Nada Assat-Anid, and Walter W. Faber Jr. Departments of Biology and Environmental Engineering, Manhattan College. Riverdale. NY 10471
In a recent study, we used the total coliform membrane filtration analysis on treated wastewater samples, and confirmed 5% of the positive colony growth with lauryl tryptose broth and brilliant green bile broth, following standard methods. The results of that study indicated a lack of confirmation in upwards of 50% of the tested positive colonies. This led to further investigation of the potential for false negative and false positive results with the confirmation procedure. Pure cultures of coliform and non-coliform bacteria have been isolated and re-isolated four times to maintain axenic cultures. These cultures were diluted in laboratory water samples, enumerated, and run through the total coliform membrane filtration analysis. Positive growth was confirmed by the same confirmation procedures, and colonies were confirmed on a Vitek identification system to verify the species of bacteria. Preliminary results will be presented.

CHARACTERIZATION OF HUMAN EYA2, A HOIVIOLOG OF THE DROSOPHIL4 EYES ABSENT GENE.
Christina A. Doyle and Brian E. Fee. Department of Biology, Manhattan College/College of Mount Saint Vincent. Riverdale. NY 10471
The Drosophila eyes absent (eya) gene is required in the development of the compound eye: without it, progenitor cells in the eye imaginal disc undergo programmed cell death (apoptosis), resulting in eyeless flies. Homologs have been found in organisms from plants to vertebrates. In humans, a family of four homologs, EYA 1-4. has been isolated, if EYA has a functional homology to Drosophila eya, it may prevent cell death, and therefore, could lead to the uncontrolled cell growth seen in cancer. Our previous research of EYA2 in human neuroblastoma cells identified four mRNA transcripts differing only at the 5' end. one of which contained a start codon fifteen bases upstream of the accepted start codon. To determine the actual 5' end of the EYA2 mRNA from a nontumorigenic tissue, we isolated the mRNA from human eye. The 5' end of the EYA2 cDNA was sequenced and found to contain the upstream start codon stated above, indicating that the human EYA2 protein sequence is five amino acids longer than previously published.

HOW ARE ORGANISMS PUT TOGETHER? WHAT CAN WE LEARN FROM THE STRUCTURES OF PLANTS?
Elvira Liclican. Laboratory of Plant Morphogenesis, Manhattan College. Riverdale. N.Y. 10471
All higher organisms have a complex structure in which appendages are connected io the main body. What are the characteristics of stresses and tissues that resist these stresses". Cactaceae are a diverse group of plants with a wide variety of morphologies and reproductive strategies. Many species have segmented stems in which terminal cladodes may be separated from main stem cladodes with varying amounts of resistance. Terminal cladodes that are removed with little resistance may form adventitious roots easily to produce new plants asexually, while other species with high resistances to cladode removal may predominately reproduce sexually. The purpose of the present study was to (1) quantitaiively determine the various stresses at joints between stem segments of two species of Opunna (0. fulglda ["jumping cholla"] and 0. verslcoior). (2) to determine iflignified cells of the xylem cells in joints provide the ma)or source of resistance to these stresses and (3) to determine if these resistances are related to whether species reproduction is correlated with resistan ce stresses. Cladodes from four branches of each of the two cactus species were taken from plants in the desert near Tucson. AZ in June 1998. Data of the present study show the following: 1) The computer-aided design package coupled with a geometric evaluation of relative positions of cladodes provides adequate estimates of joint stress parameters among cladodes. 2) bending stresses at joints were more than 10 times greater than any other stresses. 3) Expressing joint stress as a function of area of lignified xylem cell is a valid way of expressing the mechanics of joint integrity, 4) Stresses at joints as a function of area of lignified xylem cells were about four times greater for 0. julglda than for 0. versicolor. 5) The relatively high bending stress values atjoints of 0. Julglda coincides with its ability to have terminal cladodes removed easily and its ability to reproduce asexually via rooted cladodes, 6) In contrast, lower bending stress values at joints of 0. verstcolor coincide with its greater ability to retain cladodes and its characteristic to reproduce predominately via seeds I and not asexually via rooted cladodes).

BIOLOGICAL EFFECTS OF OZONE LAYER DEPLETION.
Marigrace Lim. Biology Department. Manhattan College/College of Mount Saint Vincent. Riverdale. N.Y. 10471
The thickness of the straiospheric ozone layer is sensitive to human pollutants. Data have shown that the thickness of this layer is smaller in the polar zones now than in frmer times. However, no generally accepted data indicate ihinnina in areas other (hail polar Mnes. We propose using long-lived columnar cacti as biological indicalors of zone layer depletion. The purpose of this study was to determine if surface iniuries and premature death of tall. long-lived columnar cacti have increased over the past decades and to determine if UV-B light exposures may be the causative agent for these surface injuries. The results of analyses of surface injuries from old, published photographs show that tall. long-lived, saguaro cacti have more surface injuries in the last few decades than in previous decade. These photographs show 51% increase in surface injuries over of period of fifty years. This time period coincides with the increase in the amount of UV-B light, the most deleterious component of sunlight that has been entering the earth's atmosphere. In addition, this study has also shown correlations between surface injuries and microscopic changes in the epidermis of cacti. As injuries progress from no visible injury to barking, numbers of visible stomata decreased and numbers of epidermal cell layers increased. For example, the number of crustaed epidermal layers increased from 0.0 in samples with no visible injuries to 1 1.5 layers in samples that exhibited barking the predominant visible injury symptom of cacti prior to premature death). Exposure of experimental saguaros to different types and levels of UV light demonstrated that UV-B light causes proliferation of epidermal layers and occlusion of stomata. Between 7 and 22% of stomata were occluted for UV-B exposed surfaces compared with 42 to 53% for control surfaces. There was a mean value of 2.6 epiderml cells for surfaces exposed to UV-B-while control surfaces had 1.7-1.9 cell layers on average. UV-A light has no effect on saguaro surfacesl. The tall, long-lived cacti of the Americas have been exposed to ambient sunlight for over a century. It is anticipated that continued exposure of our experimental saguaros to controlled UV-B would result in additional epidermal cell layers that will become crushed and eventually form a visible bark. The results of our experiment support our hypothesis that UV-B is the causative agent of barking and premature death of cacti, a condition that is prevalent throughout the Americas.

UNDERSTANDING CHARACTERISTICS OF STEM CELL POPULATIONS USING PLANT ROOT
MERISTEMS.
Jacklyn Pancrudo and Karen Lugrazon. Biology Depanment, .Manhattan College/College of' Mount Saint Vincent. Riverdale.N.Y. 10471
Cancer is caused by changes in cell cycle parameters in stem cell populations in both animals and plants. Plant root meristems have been used tor at least five decades as examples of cell cycle changes to stem cell populations in cancers. The present study used root meristems of several plant species to understand cell cycle parameters within stem cell populations themselves. Since (1) cells of plants do not slide relative to one another. (2) the length of cells in cell files is a reflection of the cell cycle duration (so that longer cells have longer cell cycle durations while shorter cells have shorter cell cycle times), then differences in cell lengths can be used to determine differences in lengths of ceil cycle durations within a stem cell population. Many botanists have assumed that all the cells in the terminal meristem are proliferacive and that the tissues expand exponentially. This research tested the hypothesis that changes in cell length do not occur within the first 40 cells of the root meristem for both cortex and stele cells tiles. Data of three plant species show that very marked differences in cell lengths can occuf within 5 to 15 cells from the original first five cells of files and: that stele cells show more cell enlargement than cortex cells. Each of the three plant species tested showed markedly different cell length characteristics. In this way. the data reject the above hypothesis and demonstrate that cell cycle durations change markedly for cells in the stem cell populations. In addition, the marked differences among the three species demonstrate that no generalized pattern occurs in plants. The implications ot 'these data to cancer cells in animals will be discussed.

HISTONE DEACETYLASE INHIBITORS: CONTROL OF CELL PROLIFERATION VIA POST-TRANSLATIONAL MODIFICATION RATHER THAN TRANSCRIPTIONAL REPRESSION.
Jessica Papile, James Murphy, and William A. Tramontano. Department of Biology, Manhattan College/College of Mount Saint Vincent. Riverdale. NY 10471
Butyric acid is a short-chained fatty acid that halts mitosis at concentrations of O.ImM when added to cultured root meristems of Pisum salivum. Butyrate can also act as a histone deacetylase (HDA) inhibitor. The HDA inhibitor trichosiatin A (TSA) also halts mitosis in cultured root meristems of Pisum sativum at O.OImM. Northern analysis of root meristems treated with the HDA inhibitors TSA and butyrate indicate that non-proliferating cells express notable amounts of transcripts of four, known cell proliferation associated genes. These results indicate that the HDA inhibitors butyrate and TSA halt mitosis without down-regulating the cell proliferation genes: cycA2:l, cdc2, histone Hzi-\, and MAP kinase. These genes ordinarily have low or non-existent expression levels in non-proliferating cells. Western analysis ofHDA- inhibitor treated cells reveals an increase in the level of nuclear protein acetylation at MW 13, 45, and 60. Therefore, these results suggest that the HDA inhibitors butyrate and TSA alter nuclear protein acetylation levels which may prevent the cells from dividing, without down-regulating genes typically associated with dividing cell populations.

CHARACTERIZATION OF AN ANTIBODY DIRECTED AGAINST NEURAL CELL ADHESION MOLECULES (N-CAMs).
Anne Paul. Department of Biology, Manhattan College/College of Mount Saint Vincent, Riverdale. N.Y. 10471
N-CAMs are cell surface glycoproteins with three distinct isoforms of 180, 140 and 120 Kdal. These proteins are members of the super immunoglobulin family that are involved in cell-to-cell recognition and interactions within the nervous system. In this particular study, N-CAM expression was examined in a variety of membranes including red blood cells, whole cerebellum extracts, axolemma and myelin. The proteins from these membranes were resolved utilizing two-dimensional gel electrophoresis, and transferred to nitrocellulose paper, and probed with a commercially available polyclonal anti-rat antibody (Chemicon #AB1505). Analysis of the Western Blots revealed the three isoforms of N-CAMs in cerebellum and axolemma. Association of N-CAMs with axolemma suggested probable interaction of the neuron with adjoining cells that are largely astrocyte. In contrast, the red blood cell membranes served as the negative control dnd illustrated the absence of N-CAMs. Myelin blots detected a protein with a molecular weight of 100 Kdal suggesting that the protein is a related cell adhesion molecule, specifically the myelin-associated glycoprotein (MAG). This indicated that the antibody cross-reacts with both N- CAMs and MAG.

THE UNUSUAL CHONDRIOME OF CHLAMYDOMONAS ACIDOPHILA
Pascale Rabbah and Ioanna Visviki. Department of Biology, Manhattan College/College of Mount Saint Vincent. Riverdale. N.Y. 10471
Chlamydomonas acidophila is a Linicellular. flagellated green alga of the Order Volvocales, Family Chlamydomonadaceae. It has been isolated from lakes and acidic bogs with pH as low as 2. The ultrastructure of this chlorophyte is unique, due to the large size and position of its mitochondria. The mitochondrial volumes of other Chlamydomonas species vary between 1-3% and the mitochondria are located directly below the cell membrane. The mitochondrial volume of C. acidophilia varies between 4- 6% and the mitochondria are located below the chloroplast. Examination of the chondriome, quantified by morphometric analysis, indicates that it is a dynamic cellular component, changing ontinuously during the light cycle via fragmentation, fusion and autolysis. At the onset of the light cycle (L0-L4) small mitochondria predominate. At L6-L8 the average number of mitochondrial segments decreases significantly and giant mitochondria appear via fusion, bearing clear foci and disorganized cristae. During L9-L12); the average number of mitochondrial segments increases significantly and small mitochondria are prevalent. The significance of these results is discussed and comparisons are made with other Chlamydomonas congeners.

RECOVERY OF RECOMBINANT INFLUENZA B VIRUSES FROM cDNA
Dmitny Zamarin, Kai-i Anderson. Jason Paragas and Peter Palese. Department of Microbiology, Mount Sinai School of Medicine. New York. NY 10029 and Department of Biology. Manhattan College/College of Mount Saint Vincent. Riverdale.NY10471
Recent advances in virology and understanding of structure and replication of the negative-strand RNA viruses have led to development of new methods of manipulation of viral genomes, such as rescue of synthetic genes into influenza viruses. Currently, we are working on the development of a system that would allow the rescue of infectious influenza B viruses entirely from cloned cDNA. The complete genome ofB/Yamagata/73 influenza virus was cloned into plasmids between the human RNA polymerase I promoter and hepatitis delta virus ribozyme sequence. Transfection of these plasmids into human embryonic kidney cells (293T) along with vectors expressing viral polymerase proteins results in generation in vivo of viral ribonucleoprotein (RNP) complexes, which upon expression form viral particles. While no functional viruses were detected after tranfection. we were able to show that the generated viral particles were capable of packaging and delivering a foreign reporter gene (CAT) to other cells. This plasmid-based reverse genetics technique will facilitate the study of viral structure and replication, and could lead to development of new vaccines and gene therapy vectors.