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.
Manhattan College 2000 students
Best Biology Paper - Elvira Liclican
Best Oral in Biology - Marigrace Lim
Best Oral in Molecular Biology -
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Diane Craft and Michael Judge
Dimitra Doupis, Nada Assat-Anid, and Walter W. Faber Jr
Christina A. Doyle and Brian E. Fee
Jacklyn Pancrudo and Karen Lugrazon
Jessica Papile, James Murphy, and William A. Tramontano
Pascale Rabbah and Ioanna Visviki
Dmitny Zamarin, Kai-i Anderson, Jason Paragas, and Peter
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
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.
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.
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
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.
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).
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.
CHARACTERISTICS OF STEM CELL POPULATIONS USING PLANT ROOT
Jacklyn Pancrudo and Karen Lugrazon.
Biology Depanment, .Manhattan College/College of' Mount Saint Vincent. Riverdale.N.Y.
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.
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.
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
UNUSUAL CHONDRIOME OF CHLAMYDOMONAS ACIDOPHILA
Pascale Rabbah and Ioanna Visviki.
Department of Biology, Manhattan College/College of Mount Saint Vincent. Riverdale. N.Y.
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
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
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.