Cell and Developmental Biology Commons^™

Micrornas: Tiny Regulators With Great Potential, Victor Ambros

Victor R. Ambros

Animal genomes contain an abundance of small genes that produce regulatory RNAs of about 22 nucleotides in length. These microRNAs are diverse in sequence and expression patterns, and are evolutionarily widespread, suggesting that they may participate in a wide range of genetic regulatory pathways.

Manipulation Of Prenatal Hormones And Dietary Phytoestrogens During Adulthood Alter The Sexually Dimorphic Expression Of Visual Spatial Memory, Edwin D. Lephart, Trent D. Lund

Faculty Publications

In learning and memory tasks, requiring visual spatial memory (VSM), males exhibit higher performance levels compared to females (a difference attributed to sex steroid hormonal influences). Based upon the results from our companion investigation, this study examined the influence of prenatal sex steroid hormone manipulations on VSM in adulthood, as assessed in the radial arm maze. Additionally, the influence of dietary soy phytoestrogens (i.e., the presence of high or low estrogen-like compounds present in the animal's diet) on VSM was examined in combination with the prenatal hormonal manipulations. Results: Radial arm maze performance on a phytoestrogen-rich diet: 1) females treated …

Visual Spatial Memory Is Enhanced In Female Rats (But Inhibited In Males) By Dietary Soy Phytoestrogens, Edwin D. Lephart, Trent D. Lund, Timothy W. West, Lilyan Y. Tian, Lihong H. Bu, Daniel L. Simmons, Kenneth Setchell, Herman Adlercreutz

Faculty Publications

In learning and memory tasks, requiring visual spatial memory (VSM), males exhibit superior performance to females (a difference attributed to the hormonal influence of estrogen). This study examined the influence of phytoestrogens (estrogen-like plant compounds) on VSM, utilizing radial arm-maze methods to examine varying aspects of memory. Additionally, brain phytoestrogen, calbindin (CALB), and cyclooxygenase-2 (COX-2) levels were determined. Results: Female rats receiving lifelong exposure to a high-phytoestrogen containing diet (Phyto-600) acquired the maze faster than females fed a phytoestrogen-free diet (Phyto-free); in males the opposite diet effect was identified. In a separate experiment, at 80 days-of-age, animals fed the Phyto-600 …

An Extensive Class Of Small Rnas In Caenorhabditis Elegans, Rosalind Lee, Victor Ambros

Victor R. Ambros

The lin-4 and let-7 antisense RNAs are temporal regulators that control the timing of developmental events in Caenorhabditis elegans by inhibiting translation of target mRNAs. let-7 RNA is conserved among bilaterian animals, suggesting that this class of small RNAs [microRNAs (miRNAs)] is evolutionarily ancient. Using bioinformatics and cDNA cloning, we found 15 new miRNA genes in C. elegans. Several of these genes express small transcripts that vary in abundance during C. elegans larval development, and three of them have apparent homologs in mammals and/or insects. Small noncoding RNAs of the miRNA class appear to be numerous and diverse.

Mutations In Sid2, A Novel Gene In Saccharomyces Cerevisiae, Cause Synthetic Lethality With Sic1 Deletion And May Cause A Defect During S Phase, Matthew D. Jacobson , '97, Claudia C. Muñoz , '99, Kirstin S. Knox , '99, Beth E. Williams , '01, Lenette L. Lu , '02, Frederick Cross , '78, Elizabeth Ann Vallen

Biology Faculty Works

SIC1 encodes a nonessential B-type cyclin/CDK inhibitor that functions at the G1/S transition and the exit from mitosis. To understand more completely the regulation of these transitions, Mutations causing synthetic lethality with sic1 Delta were isolated. In this screen, we identified a novel gene, SID2, which encodes an essential protein that appear., to be required for DNA replication or repair. sid2-1 sic1 Delta strains and sid2-21 temperature-sensitive strains arrest preanaphase as large-budded cells with a single nucleus, a short spindle, and an similar to 2C DNA content. RAD9, which is necessary for the DNA damage checkpoint, is required for the …

Development. Dicing Up Rnas, Victor Ambros

Victor R. Ambros

Comment on: A cellular function for the RNA-interference enzyme Dicer in the maturation of the let-7 small temporal RNA. [Science. 2001]

A Genetic And Molecular Analysis Of Spermiogenesis Initiation In Caenorhabditis Elegans, Paul J. Muhlrad

Paul J Muhlrad

How do cells acquire and regulate their specialized forms and functions? This is the fundamental question underlying the experiments described in this dissertation. To examine this question, I have peered deeply into an exquisitely specialized cell from a genetically tractable organism: the spermatozoon of the nematode, Caenorhabditis elegans. C. elegans sperm orchestrate a fantastic morphogenetic transformation under severe constraints of time and cellular resources. In response to an external signal, spermatids reorganize their membranes and cytoskeleton to form crawling spermatozoa.This maturation, termed spermiogenesis, ensues without any new gene expression. To better understand this signaling pathway, I isolated suppressors of a …

A Novel Strategy Utilizing Co-Expression Of Murine Il-12 And Antisense Tgf-Pl Against H238 Tumor Formation, Craig A. Seheult

Loma Linda University Electronic Theses, Dissertations & Projects

Cytokines have a profound effect on immune modulation, thus playing a significant role in cancer gene therapy. A variety of human carcinomas secrete transforming growth factor-beta (TGF-β), a cytokine with potent immunosuppressive properties. Suppressing TGF-β could be a key to successfully combating malignancies, such as gliomas, mammary, and colon cancers, that over-express TGF-β. Conversely, interleukin-12 (IL-12) is a potent immunostimulatory cytokine capable of regulating T and NK cell-mediated cytotoxic responses (Trinchieri, 1994) during an anti-tumor response. Activation of the immune system with such immunostimulatory cytokines renders it inert to the suppressive effects of TGF-β. To better understand these processes our …

Effects Of Liposome-Encapsulated Tnf-Alpha And Radiation Against Colon Cancer, Dong Won Kim

Loma Linda University Electronic Theses, Dissertations & Projects

Previous studies have shown that tumor necrosis factor-α (TNF-α) can augment the antitumor effects of radiation, inhibiting tumor progression to a greater extent than either modality alone. However, nonspecific toxicities associated with TNF-α have limited its use as an adjuvant. This limitation is potentially resolved with the encapsulation of TNF-α in sterically stabilized, PEGylated liposomes (STEALTH®). Thus, the overall hypothesis of the research was that the combination of radiation with STEALTH® liposome encapsulated TNF-α (SL-TNF-α) will have greater efficacy and fewer dose-limiting side effects in the LS174T human colorectal tumor xenograft model compared to the combination of free TNF-α and …

Identification Of Functional Amino Acids In The G Protein Alpha-Subunit, Robert E. Gundersen

University of Maine Office of Research Administration: Grant Reports

Guanine nucleotide-binding proteins (G proteins) are vital to a wide variety of physiological processes such as vision, hormone response, olfaction, and development. G protein-mediated signal transduction involves a complex network of receptors and second messenger pathways, and proper regulation of the network is essential for proper response of the system. A detailed analysis of the functional domains of the G protein alpha-subunit has been initiated by screening a library of random mutations in the subunit, G alpha2, of the slime mold, Dictyostelium. The function of G alpha2 function is essential for the developmental life cycle of this organism. It regulates …

Positive Regulation Of Pka On Human Gonadotropin-Releasing Hormone (Hgnrh) Gene Expression In Human Placental Jeg-3 Cells, Zhaoyang Wen

Chemistry & Biochemistry Theses & Dissertations

Using the human placental choriocarcinoma JEG-3 cell line as an in vitro human placental model, we studied the mechanisms of the PKA positive regulation of the hGnRH gene expression in the human placenta. Studies in JEG3 cells showed that through the PKA catalytic subunit a, human GnRH upstream promoter activity was stimulated by PKA signaling pathway in a cAMP dependent mechanism. The sequence between —202 (Afl II) and —554 (BamH I) base pair in the human GnRH upstream promoter region appeared to be responsible for the PKA positive regulation of the gene expression. Furthermore, Western blot analysis demonstrated the involvement …

Paterson's Curse Management Handbook, Amelia Mclarty, Errol Kruger, Peter Stubbs, John Peirce, Chris Hawkins, Paul Wilson, Simon Merewether

Bulletins 4000 -

Paterson’s curse (Echium plantagineum) is one of the most damaging weeds to the Australian meat and wool industries. It infests an estimated 33 million hectares in southern Australia, at an annual cost to the sheep industry of $250 million in lost pasture productivity, control costs and wool contamination. In WA, it is found on about 5000 agricultural properties infesting some 500,000 ha. It is an extremely invasive weed, that reduces pasture productivity and stock carrying capacity by competing with and excluding more beneficial pasture species. The weed is also toxic to stock. It is important to implement control measures on …

Association Of Mesenchymal Cells And Immunoglobulins With Differentiating Epithelial Cells, Antonin Bukovsky, Michael R. Caudle, Jeffrey A. Keenan, Nirmala B. Upadhyaya, Stuart E. Van Meter, Jay Wimalsena, Robert F. Elder

Faculty Publications and Other Works -- General Biology

Background

Mesenchymal-epithelial interactions play an important role in the physiology and pathology of epithelial tissues. Mesenchymal cells either associate with epithelium basement membrane [pericytes and perivascular monocyte-derived cells (MDC)] or reside within epithelium (MDC and T cells). Although intraepithelial mesenchymal cells were suggested to contribute to the epithelium physiology, their association with particular steps in differentiation of epithelial cells, interactions among themselves, and their fate remain unclear. We studied epitopes of mesenchymal cells and their products (immunoglobulins) in stratified epithelium of uterine ectocervix, which is one of the prototypes of complete cellular differentiation from stem into the aged cells.

Results …

Salivary Acinar Cells From Aquaporin 5-Deficient Mice Have Decreased Membrane Water Permeability And Altered Cell Volume Regulation, Carissa M. Krane, James E. Melvin, Ha-Van Nguyen, Linda Richardson, Jennifer E. Towne, Thomas Doetschman, Anil G. Menon

Biology Faculty Publications

Aquaporins (AQPs) are channel proteins that regulate the movement of water through the plasma membrane of secretory and absorptive cells in response to osmotic gradients. In the salivary gland, AQP5 is the major aquaporin expressed on the apical membrane of acinar cells. Previous studies have shown that the volume of saliva secreted by AQP5-deficient mice is decreased, indicating a role for AQP5 in saliva secretion; however, the mechanism by which AQP5 regulates water transport in salivary acinar cells remains to be determined. Here we show that the decreased salivary flow rate and increased tonicity of the saliva secreted byAqp5 …

The Wings Of Bombyx Mori Develop From Larval Discs Exhibiting An Early Differentiated State: A Preliminary Report, Madhuri Kango-Singh, Amit Singh, K. P. Gopinathan

Biology Faculty Publications

Lepidopteran insects present a complex organization of appendages which develop by various mechanisms. In the mulberry silkworm, Bombyx mori, a pair of meso- and meta-thoracic discs located on either side in the larvae gives rise to the corresponding fore- and hind-wings of the adult. These discs do not experience massive cell rearrangements during metamorphosis and display the adult wing vein pattern. We have analysed wing development in B. mori by two approaches, viz., expression of patterning genes in larval wing discs, and regulatory capacities of larval discs following explantation or perturbation. Expression of Nubbin is seen all over the presumptive …

Quantification Of Benzo[A]Pyrene-Guanine Adducts In In Vitro And In Vivo Tissue Samples By Lc Tandem Mass Spectrometry, Po-Chang Chiang

Dissertations

No abstract provided.

Morphine Enhances The Permeability Across Vascular Endothelial Cell Barriers, Jenine Anday

Seton Hall University Dissertations and Theses (ETDs)

Using three types of in vitro endothelial cell barrier models, we investigated the direct effects and mechanisms of morphine on vascular permeability. Our current studies illustrate LPS induces permeability across these VEC barriers, and is significantly enhanced when co-treated with morphine, displayed using the [ 14 CJ-inulin paracellular marker. Incubation with morphine alone induces permeability in a concentration­ depeodent manner, and is not blocked by the addition of naloxone. Morphine enhances the detrimental effects of LPS on cell viability and alone also decreases endothelial cell viability, concentration-dependently, which is also not affected by naloxone, as demonstrated by the trypan blue …

Reversible Calcium-Regulated Stopcocks In Legume Sieve Tubes, Michael Knoblauch, Winfried Peters, Katrin Ehlers, Aart Van Bel

Winfried S. Peters

Evidence For Two Mechanisms Of Amino Acid Osmolyte Release From Hippocampal Slices, R. Franco, M. E. Torres-Marquez, H. Pasantes-Morales

School of Veterinary and Biomedical Sciences: Faculty Publications

A 30% decrease in osmolarity stimulated ³H-taurine, ³H-GABA and glutamate (followed as ³H-D-aspartate) efflux from rat hippocampal slices. ³H-taurine efflux was activated rapidly but inactivated slowly. It was decreased markedly by 100 μM 5-nitro-(3-phenylpropylamino) benzoic acid (NPPB) and 600 μM niflumic acid and inhibited strongly by tyrphostins AG18, AG879 and AG112 (25–100 μM), suggesting a tyrosine kinase-mediated mechanism. Hyposmolarity activated the mitogen-activated protein kinases (MAPK) extracellular-signal-related kinase-1/2 (ERK1/ERK2) and p38, but blockade of this reaction did not affect ³H-taurine efflux. Hyposmosis also activated phosphatidyl-inositide 3-kinase (PI3K) and its prevention by wortmannin (100 nM) essentially abolished …

Does Growth Correlate With Turgor-Induced Elastic Strain In Stems? A Re-Evaluation Of De Vries' Classical Experiments, Winfried Peters, Maggie Farm, A. Kopf

Winfried S. Peters

Nuclear Export Of 60s Ribosomal Subunits Depends On Xpo1p And Requires A Nuclear Export Sequence-Containing Factor, Nmd3p, That Associates With The Large Subunit Protein Rpl10p, Olivier Gadal, Daniela Strau, Jacques Kessl, Bernard Trumpower

Dartmouth Scholarship

Nuclear export of ribosomes requires a subset of nucleoporins and the Ran system, but specific transport factors have not been identified. Using a large subunit reporter (Rpl25p-eGFP), we have isolated several temperature-sensitive ribosomal export (rix) mutants. One of these corresponds to the ribosomal protein Rpl10p, which interacts directly with Nmd3p, a conserved and essential protein associated with 60S subunits. We find that thermosensitive nmd3 mutants are impaired in large subunit export. Strikingly, Nmd3p shuttles between the nucleus and cytoplasm and is exported by the nuclear export receptor Xpo1p. Moreover, we show that export of 60S subunits is Xpo1p dependent. We …

Chromosome Movement In Mitosis Requires Microtubule Anchorage At Spindle Poles, Michael B. Gordon, Louisa Howard, Duane A. Compton

Dartmouth Scholarship

Anchorage of microtubule minus ends at spindle poles has been proposed to bear the load of poleward forces exerted by kinetochore-associated motors so that chromosomes move toward the poles rather than the poles toward the chromosomes. To test this hypothesis, we monitored chromosome movement during mitosis after perturbation of nuclear mitotic apparatus protein (NuMA) and the human homologue of the KIN C motor family (HSET), two noncentrosomal proteins involved in spindle pole organization in animal cells. Perturbation of NuMA alone disrupts spindle pole organization and delays anaphase onset, but does not alter the velocity of oscillatory chromosome movement in prometaphase. …

Erv41p And Erv46p New Components Of Copii Vesicles Involved In Transport Between The Er And Golgi Complex, Stefan Otte, William J. Belden, Matthew Heidtman, Jay Liu, Ole N. Jensen, Charles Barlowe

Dartmouth Scholarship

Proteins contained on purified COPII vesicles were analyzed by matrix-assisted laser desorption ionization mass spectrometry combined with database searching. We identified four known vesicle proteins (Erv14p, Bet1p, Emp24p, and Erv25p) and an additional nine species (Yip3p, Rer1p, Erp1p, Erp2p, Erv29p, Yif1p, Erv41p, Erv46p, and Emp47p) that had not been localized to ER vesicles. Using antibodies, we demonstrate that these proteins are selectively and efficiently packaged into COPII vesicles. Three of the newly identified vesicle proteins (Erv29p, Erv41p, and Erv46p) represent uncharacterized integral membrane proteins that are conserved across species. Erv41p and Erv46p were further characterized. These proteins co- localized to …

Protein Targeting And Translocation, Barry D. Bruce, D.A. Phoenix

Faculty Publications and Other Works -- Biochemistry, Cellular and Molecular Biology

No abstract provided.

A Role For Actin, Cdc1p, And Myo2p In The Inheritance Of Late Golgi Elements In Saccharomyces Cerevisiae, Olivia W. Rossanese, Catherine A. Reinke, Brooke J. Bevis, Adam T. Hammond, Irina B. Sears, James O'Connor, Benjamin S. Glick

Faculty Publications

In Saccharomyces cerevisiae, Golgi elements are present in the bud very early in the cell cycle. We have analyzed this Golgi inheritance process using fluorescence microscopy and genetics. In rapidly growing cells, late Golgi elements show an actin-dependent concentration at sites of polarized growth. Late Golgi elements are apparently transported into the bud along actin cables and are also retained in the bud by a mechanism that may involve actin. A visual screen for mutants defective in the inheritance of late Golgi elements yielded multiple alleles of CDC1. Mutations in CDC1 severely depolarize the actin cytoskeleton, and these …

Abnormal Expression Of P27kip1 Protein In Levator Ani Muscle Of Aging Women With Pelvic Floor Disorders – A Relationship To The Cellular Differentiation And Degeneration, Antonin Bukovsky, Pleas Copas, Michael R. Caudle, Maria Cekanova Ms, Rndr, Phd, Tamara Dassanayake, Bridgett Asbury, Stuart E. Van Meter, Robert F. Elder, Jeffrey B. Brown, Stephanie B. Cross

Maria Cekanova MS, RNDr, PhD

Background Pelvic floor disorders affect almost 50% of aging women. An important role in the pelvic floor support belongs to the levator ani muscle. The p27/kip1 (p27) protein, multifunctional cyclin-dependent kinase inhibitor, shows changing expression in differentiating skeletal muscle cells during development, and relatively high levels of p27 RNA were detected in the normal human skeletal muscles.

Methods Biopsy samples of levator ani muscle were obtained from 22 symptomatic patients with stress urinary incontinence, pelvic organ prolapse, and overlaps (age range 38–74), and nine asymptomatic women (age 31–49). Cryostat sections were investigated for p27 protein expression and type I (slow …

Protein Targeting And Translocation, Barry D. Bruce, D.A. Phoenix

Barry D. Bruce

No abstract provided.

Analysis Of E. Coli Dna From Selected Source(S) Using Pulsed-Field Gel Electrophoresis, Sheema Doshi

Theses, Dissertations and Capstones

The presence of fecal coliforms in surface waters is a public health issue throughout the United States and Canada. The presence of Escherichia coli (E. coli) indicates recent fecal contamination from either animal or human sources. To solve this growing problem one must identify the source(s) of such contamination by creating a database of DNA profiles of£. coli isolates. Hence, tracking the source(s) of E. coli can help link water contamination to animal source(s) of contamination. In this study, pulsed field gel electrophoresis (PFGE) was used to analyze DNA from E. coli isolates from known animals. For PFGE, the bacterial …

Genomic Diversity Of Pseudomonas Aeruginosa In Cystic Fibrosis, Nathaniel Edwards Head

Theses, Dissertations and Capstones

Pseudomonas aeruginosa is a common environmental microorganism. However, it has the genetic capacity to cause diseases in patients with compromised host defense systems. Cystic fibrosis (CF) is one of the major hereditary diseases among Caucasian populations. CF patients are born with a defective chloride channel that is responsible for maintaining fluid and electrolyte balance across the lumen of the lung. This imbalance leads to the production of an abnormal dehydrated viscous mucous, prohibiting the patient from normally clearing the respiratory airway. Chronic pulmonary infections with P. aeruginosa are the major causes of high morbidity and mortality in CF. Two of …

Factors Influencing Spatial Variability In Soil Nitrogen (N) Dynamics In N-Treated And Untreated Watersheds Of The Fernow Experimental Forest, West Virginia, Nikki Lenore Lyttle

Theses, Dissertations and Capstones

The central Appalachian region of the United States receives some of the highest inputs of nitrogen (N) due to acidic deposition in the nation. It is believed that these high could levels contribute to a decline in forest soils within the next 50 to 70 yrs. This study examines factors that influence spatial variability in N-treated and untreated watersheds of the Fernow Experimental Forest, Parsons, West Virginia. Within each of the two watersheds [WS4 untreated control, > 100 yr.; WS3 N-treated, acidified, clear cut, ~ 31 yr.], two 0.04 ha plots, one high N and one low N, were selected for …