Cell Biology Commons^™

Efficacy Of AllowashTm Solution In Solubilization Of Bone Marrow, Alyce Marie Linthurst

Biological Sciences Theses & Dissertations

LifeNet currently cleans allograft bone utilizing Allowash^TM Technology, which encompasses the use of a closed continuous circulation chamber to remove bone marrow by flushing it with Allowash^TM Solution for a prescribed time and temperature after which, it is rinsed with various solutions to remove residual detergent that could be toxic to the recipient. A portion of this process employs Allowash^TM Solution to improve the solubilization and removal of protein and cellular elements in the red bone marrow. This study examines the effectiveness of Allowash^TM Solution in comparison to its component detergents Nonidet P40, Nonoxynol 9 and …

Sec35p, A Novel Peripheral Membrane Protein, Is Required For Er To Golgi Vesicle Docking, Susan M. Vanrheenen, Xiaochun Cao, Vladimir V. Lupashin, Charles Barlowe, M. Gerard Waters

Dartmouth Scholarship

SEC35 was identified in a novel screen for temperature-sensitive mutants in the secretory pathway of the yeast Saccharomyces cerevisiae (Wuestehube et al., 1996. Genetics. 142:393–406). At the restrictive temperature, the sec35-1 strain exhibits a transport block between the ER and the Golgi apparatus and accumulates numerous vesicles. SEC35 encodes a novel cytosolic protein of 32 kD, peripherally associated with membranes. The temperature-sensitive phenotype of sec35-1 is efficiently suppressed by YPT1, which encodes the rab-like GTPase required early in the secretory pathway, or by SLY1-20, which encodes a dominant form of the ER to Golgi target …

Confocal Microscopy: A Powerful Tool For Biological Research, Amit Singh, K. P. Gopinathan

Biology Faculty Publications

Conventional light microscopy allows the observation of living as well as fixed cells and tissues to generate two-dimensional images. The out-of-focus information often obscures the ultrastructural details, especially in thick specimens with overlapping structures. The earliest available light microscopy visualized the objects in hydrated state in two-dimensions during their temporal development. The emergence of electron microscopy (EM) provided superb resolution of ultrastructural details, but it was applicable only for objects in the dehydrated state and thereby potentially introducing handling artifacts. The usefulness of optical methods, however, has been limited by the poor depth discrimination. Often, the fluorescence and reflectance images …

The Biology Of Xenopus By R. C. Tinsley And H. C. Kobel, Rafael O. De Sá

Biology Faculty Publications

The Biology of Xenopus presents a summary of current knowledge about a single genus resulting from a symposium held at the Zoological Society of London in September 1992. This approach to summarizing available information has also been taken for other taxa, such as Atelopus (Lotters, 1996). However, the task of compiling data for Xenopus is enormous relative to any other amphibian group, because Xenopus laevis has become a model system for molecular and development research (Cannatella and de Sa, 1993). Unfortunately, most of our knowledge of Xenopus is biased toward this single species. There are about 20 recognized species of …

Chondrocranial Morphology Of Leptodactylus Larvae (Leptodactylidae: Leptodactylinae): Its Utility In Phylogenetic Reconstruction, Peter M. Larson, Rafael O. De Sá

Biology Faculty Publications

Chondrocranial morphology of leptodactylid frogs is scarcely known and has not been completely described for any species of Leptodactylus. We describe the diversity of chondrocranial morphology in the genus Leptodactylus based on the analysis of 22 species, representing the four species groups: the fuscus Group, ocellatus Group, melanonotus Group, and pentadactylus Group. Furthermore, 26 characters are identified and used in a phylogenetic analysis. The phylogenetic analysis using Physalaemus, Crossodactylus, and Hylodes as outgroups suggests two monophyletic clades within Leptodactylus: the melanonotus-ocellatus clade and the pentadactylusfuscus clade. However, it does not support the monophyly of the species groups as currently recognized …

Coupled Er To Golgi Transport Reconstituted With Purified Cytosolic Proteins, Charles Barlowe

Dartmouth Scholarship

A cell-free vesicle fusion assay that reproduces a subreaction in transport of pro-alpha-factor from the ER to the Golgi complex has been used to fractionate yeast cytosol. Purified Sec18p, Uso1p, and LMA1 in the presence of ATP and GTP satisfies the requirement for cytosol in fusion of ER-derived vesicles with Golgi membranes. Although these purified factors are sufficient for vesicle docking and fusion, overall ER to Golgi transport in yeast semi-intact cells depends on COPII proteins (components of a membrane coat that drive vesicle budding from the ER). Thus, membrane fusion is coupled to vesicle formation in ER to Golgi …

Recovery Of Rna Polymerase Ii Synthesis Following Dna Damage In Mutants Of Saccharomyces Cerevisiae Defective In Nucleotide Excision Repair, Michael S. Reagan, Errol C. Friedberg

Biology Faculty Publications

We have measured the kinetics of the recovery of mRNA synthesis in the inducible GAL10 and RNR3 genes after exposure of yeast cells to ultraviolet (UV) radiation. Such recovery is abolished in mutant strains defective in nucleotide excision repair (NER) of DNA, including a rad23 mutant. Mutants defective in the RAD7 or RAD16 genes, which are required for the repair of the non-transcribed strand but not the transcribed strand of transcriptionally active genes, show slightly faster recovery of RNA synthesis than wild-type strains. A strain deleted of the RAD26 gene, which is known to be required for strand-specific NER in …

Endosome To Golgi Retrieval Of The Vacuolar Protein Sorting Receptor, Vps10p, Requires The Function Of The Vps29, Vps30, And Vps35 Gene Products, Matthew N. J. Seaman, Eric G. Marcusson, Joan Lin-Cereghino, Scott D. Emr

College of the Pacific Faculty Articles

Mutations in the S. cerevisiae VPS29 and VPS30 genes lead to a selective protein sorting defect in which the vacuolar protein carboxypeptidase Y (CPY) is missorted and secreted from the cell, while other soluble vacuolar hydrolases like proteinase A (PrA) are delivered to the vacuole. This phenotype is similar to that seen in cells with mutations in the previously characterized VPS10 and VPS35 genes. Vps10p is a late Golgi transmembrane protein that acts as the sorting receptor for soluble vacuolar hydrolases like CPY and PrA, while Vps35p is a peripheral membrane protein which cofractionates with membranes enriched in Vps10p. The …

Analysis Of Gene Expression During Embryonic Development In Mulberry Silkworm Bombyx Mori, Amit Singh, K. P. Gopinathan

Biology Faculty Publications

We have developed a method for dechorionation and devitellinization of the silkworm eggs without damage, to facilitate the analysis of gene expression during embryonic development of Bombyx mori. Making use of antibodies available from heterologous systems, the spatio-temporal expression patterns of peroxidase and proliferating cell nuclear antigen have been directly visualized in whole mount embryos at various stages of development without the need for generating transformed ·lines carrying specific reporter constructs. The B. mori system, previously unamenable for such studies, could thus serve as an attractive model for molecular analysis of insect development.

The attention lavished on Drosophila melanogaster as …

In Vivo Incorporation Of Tritium To Measure Lipogenesis In Red Skeletal Muscle: Significance Of A Nonlipogenic Diet In La/N-Fa(K)("Corpulent") And Zucker Fa Rat Strains, Karin Traci Mann

Theses, Dissertations and Capstones

Skeletal muscle lipogenesis in rodents may equal that of liver but its importance in the obese animals is not known. We evaluated the rate of fatty acid and triglyceride synthesis and storage content in red skeletal muscles, liver, inguinal fat, and retroperitoneal fat of the LA/N-fa^k (“corpulent”) and Zucker fa obese rat strains. Obese-lean pairs of both rat strains, weaned at 32 days, fed a nonlipogenic diet (54% starch for 21 days) were either dosed with 2 µCi 3H-H² O per gram of body weight and tested after 60 minutes for whole body 3H-incorporation into fatty acids and …

Reversal Of Cell Fate Determination In Caenorhabditis Elegans Vulval Development, Susan Euling, Victor Ambros

Dartmouth Scholarship

In Caenorhabditis elegans, the fates of the multipotent vulval precursor cells (VPCs) are specified by intercellular signals, The VPCs divide in the third larval stage (L3) of the wild type, producing progeny of determined cell types, In lin-28 mutants, vulva development is similar to wild-type vulva development except that it occurs precociously, in the second larval stage (L2), Consequently, when lin-28 hermaphrodites temporarily arrest development at the end of L2 in the dauer larva stage, these otherwise determined VPC progeny become reprogrammed back to the multipotent, signal- sensitive state of VPCs. Our results indicate that VPC fate determination by intercellular …

The Evaluation Of Succinylcholine Chloride For Harmful Effects Upon Early-Stage Mouse Embryos, Francis Warren Prescott

Biological Sciences Theses & Dissertations

Succinylcholine (SCh) chloride is a depolarizing neuromuscular blocking agent used to paralyze patients for surgical procedures. Over 1,800 clinical observations of newborn infants have demonstrated that succinylcholine does not cross the placenta in quantities detrimental to fetuses (Kolstad, et al., 1957, Bakhoum, et al., 1957, Hodges, et al., 1959, Duffield, et al., 1958, Dennis, et al., 1954, 1956, McNab, 1955, Coleman, et al., 1956, Lund, 1953). Later investigations which involved comparison of blood levels of SCh in the mother to those in fetuses bolstered the finding that SCh fails to cross the placenta in significant amounts (Moya and Kvisselgaard, 1961, …

Mutations In Rad27 Define A Potential Link Between G₁ Cyclins And Dna Replication, Elizabeth Ann Vallen, Frederick R. Cross , '78

Biology Faculty Works

The yeast Saccharomyces cerevisiae has three G1 cyclin (CLN) genes with overlapping functions. To analyze the functions of the various CLN genes, we examined mutations that result in lethality in conjunction with loss of cln1 and cln2. We have isolated alleles of RAD27/ERC11/YKL510, the yeast homolog of the gene encoding flap endonuclease 1, FEN-1.cln1 cln2 rad27/erc11 cells arrest in S phase; this cell cycle arrest is suppressed by the expression of CLN1 or CLN2 but not by that of CLN3 or the hyperactive CLN3-2. rad27/erc11 mutants are also defective in DNA damage repair, as determined by their increased sensitivity to …

Enhancer Trap Technique: A Novel Tool For Identification And Developmental Characterization Of Genes Of Drosophila, Amit Singh

Biology Faculty Publications

The classical technique of mutational screen for identification of genes controlling early development has now approached saturation. A new era in genetic identification and developmental characterization of genes in Drosophila has commenced with the advent of the enhancer trap technique. This technique involves mobilization of a P-lacZ vector to diverse chromosomal locations in the fruit fly genome to bring it under the regulation of developmentally expressed genes or their enhancer elements. The technique offers a strikingly elegant method of gaining entry into fruit fly genes.

An Improved Method For Chemical Devitellinization Of X-Gal Stained Drosophila Embryos, Amit Singh, Madhuri Kango-Singh, P. Sinha

Biology Faculty Publications

In Drosophila developmental biological studies, X-gal staining is commonly employed to study the spatio-temporal expression of the lacZ reporter gene in the transformed flies or their embryos. Study of the lacZ pattern in embryos often suffers from the lack of an efficient and high yieldirrg technique for devitellinization of X-gal stained embryos. Devitellinization techniques employed during antibody staining, in situ hybridization or embryonic cuticular preparations generally do not give satisfactory results when used for similar purpose in X-gal stained embryos. This results in the flaky appearance of the blue stain. We present here an improved chemical devitellinization technique which gives …

Development And Application Of A Thermistor Current Meter, Carl M. Way, Albert J. Burky, Christine Miller-Way

Biology Faculty Publications

This report provides details for the construction of a hot-bead thermistor current meter which is capable of measuring water velocities on a millimeter spatial scale and for the construction of a compact and accurate calibration system. Hot-bead thermistor current meters can be built with response times of 200 ms capable of measuring velocities between 0.1 and 80 cm s-1. The construction of a sturdy probe for application in lotic systems such as high gradient Hawaiian streams was achieved by the use of heavy-duty acrylic tubing, small stainless steel gas-chromatography tubing, and flexible Tygon spaghetti tubing. An acrylic handle anchors the …

Ribosomal Dna And The Phylogeny Of Frogs, David M. Hills, Loren K. Ammerman, Michael T. Dixon, Rafael O. De Sá

Biology Faculty Publications

Phylogenetic analysis of 1656 aligned sites in the 28S ribosomal RNA gene of frogs supports some of the recently recognized higher groups of anurans but provides counter-support for others. The 28S rDNA data support the monophyly of the recently recognized Pipanura (me- sobatrachians plus neobatrachians), which in turn indicates paraphyly of archaeobatrachians. Me- sobatrachians (pelobatoids plus pipoids), which are either considered paraphyletic or weakly sup- ported as monophyletic in morphological analyses, also receive support as a monophyletic group from the 28S rDNA data. Hyloidea (=Bufonoidea), which is widely recognized but lacks morphological support, receives some molecular support as being monophyletic. …

Retention Of Cellular Viability And Growth Potential In Cryopreserved Bovine Arterial Tissue: Implications For Clinical Use Of Cryopreserved Vascular Allografts, Kip G. Gardner

Biological Sciences Theses & Dissertations

Segments of bovine calf aortas and pulmonary arteries were cryopreserved at -196° C for two weeks to 18 months and subsequently thawed. Cellular morphology, viability, and growth potential in these segments were then compared with the same attributes in segments from fresh (control) arteries. Scanning electron microscopy revealed no disruption of intimal surfaces attributable to cryopreservation. Endothelial cells were successfully cultured from all cryopreserved arterial segments. Endothelial monolayers grown from cryopreserved arteries had a cobblestone appearance and expressed Factor VIII antigen but not smooth muscle alpha-actin. Plating efficiency in primary cultures of endothelial cells was lower for cryopreserved arteries than …

Phase I Trial Of The Murine Monoclonal Anti-Gd2 Antibody 14g2a In Metastatic Melanoma, Mansoor Saleh, M B. Khazaeli, Richard Wheeler, Edward Dropcho, Tiepu Liu, Marshall Urist, Donald Miller, Sharon Lawson, Pam Dixon, Charles Russell

Haematology and Oncology, East Africa

In a phase I trial, 12 patients with G_D2 antigen-positive metastatic melanoma received the murine anti-G_D2 monoclonal antibody 14G2a. The monoclonal antibody was administered in four doses over an 8-day period with total dose ranging from 10 to 120 mg. All patients receiving >10 mg of 14G2a experienced transient abdominal/pelvic pain during the antibody infusion. Five patients had a delayed extremity pain syndrome following the third and fourth antibody infusion. Four of the five patients developed neurological toxicity, including two patients with significant although reversible motor neuropathy. Two of the patients developed hyponatremia secondary to a syndrome of …

Osteoinductivity Of Demineralized Bone: A Quantitative In Vitro Assessment, Yihong Zheng

Biological Sciences Theses & Dissertations

Acid demineralized bone (DMB) implanted in extraosseous sites induces bone formation. In vivo studies have shown that DMB stimulates the differentiation of mesenchymal cells into osteogenic cells. In this study an in vitro system was developed to demonstrate the osteoinductivity of DMB quantitatively. This in vitro working system was used to assess the osteoinductivity of variously processed DMB products as a means to evaluate their processing protocols. A human dermal fibroblastic (HDF) cell line was initiated from tissue explants as a working model, along with a human periosteal (HPO) cell line as a control. Ground DMB processed by LifeNet Transplant …

The Submembrane Machinery For Nicotinic Acetylcholine Receptor Clustering, S. C. Froehner

Dartmouth Scholarship

No abstract provided.

Fertilization And Mouse Embryo Development In The Presence Of Midazolam, Maria Leavitt

Biological Sciences Theses & Dissertations

The development of mouse embryos in vitro has provided a unique opportunity for the study of cellular differentiation and the influence of various agents on differentiation. Midazolam is administered for conscious sedation in the recovery of human ova. In the Norfolk IVF Program, midazolam is found in varying concentrations in follicular fluid. Mouse embryo formation and development has been used to test midazolam's effect on fertilization and early development as a model of human in vitro fertilization and development. Midazolam' s effects were studied using varying doses of midazolam on (1) in vivo fertilization and subsequent cell division, and (2) …

The Effects Of Glutathione On Human Keratinocyte, Manisha N. Trivedi

Theses and Dissertations in Biomedical Sciences

Cultured human keratinocytes were grown in vitro to assess the effects of glutathione, a thiol, on human keratinocyte proliferation. Single keratinocytes were harvested from newborn (greater than 5 days old) foreskin and were grown in culture flasks such that experiments could be performed in approximately 2 to 3 weeks. In order to evaluate the effects of glutathione (GSH) on keratinocyte growth, experiments were performed to determine the growth curve, plating efficiency, and incorporation of radiolabelled thymidine. Growth studies, performed with different concentrations of glutathione, revealed that 0.065 μM was the most effective concentration when compared to control cell cultures, and …

Electrophysiology, Cell Calcium, And Mechanisms Of Hepatocyte Volume Regulation, Walid E. Khalbuss

Electronic Theses and Dissertations

The electrophysiologic technique (Reuss, L., Proc. Natl. Acad. Sci. USA 82:6014, 1985) was modified to measure changes in steady-state hepatocyte volume during osmotic stress. Hepatocytes in mouse liver slices were loaded with tetramethylammonium ion (TMA$\sp{+}$) during transient exposure of cells to nystatin. Intracellular TMA$\sp{+}$ activity (a$\sp{\rm i}\sb{\rm TMA}$) was measured with TMA$\sp{+}$-sensitive, double-barreled microelectrodes. Loading hepatocytes with TMA$\sp{+}$ did not change their membrane potential (V$\sb{\rm m}$), and under steady-state conditions a$\sp{\rm i}\sb{\rm TMA}$ remained constant over 4 min in single impalements. Hyperosmotic solutions (50, 100, & 150 mM sucrose added to media) and hyposmotic solutions (sucrose in media reduced by …

Evidence Of A Neurogenic Component During Ige-Mediated Inflammation In Murine Skin, Victoria Jean Cavanaugh

Biological Sciences Theses & Dissertations

The direct stimulation of sensory nerves or the intradermal injection of neuropeptides triggers a cutaneous inflammatory response that involves the degranulation of mast cells. These neurogenic inflammatory reactions are physiologically similar to IgE-mediated hypersensitivity responses, which also depend on mast cells. Although non-antigenic stimuli distinguish neurogenic from IgE-mediated inflammation, the similarity of their effector stages suggested that peripheral nerves may also participate in lgE-mediated responses. To examine this hypothesis, IgE responses were elicited in denervated skin. A murine model was developed where the footpad was denervated by surgically removing a 2 mm segment of the sciatic nerve, proximal to the …

The Effects Of Adjacent Cell Fusion And Ultraviolet Radiation Exposure On Viral Plaque Formation With Herpes Simplex Virus Type I, Rebecca Conner

Masters Theses & Specialist Projects

In mammalian cell-virus systems, it has been observed that damage caused by exposure of the cell to ultraviolet radiation (UV) will result in an increase in viral plaque development rate. This phenomenon is termed the Large Plaque Effect (LPE). Apparently, viral plaque development increases at a faster rate for Herpes Simplex Virus (HSV) when it is assayed on certain UV-irradiated mammalian cells. The consequence of this increase in plaque development rate is that viral plaques appear larger on irradiated monolayers of cells when compared to plaques that developed on unirradiated cellular monolayers.

The cause of the LPE is not yet …

Atp-Dependent Formation And Motility Of Aster-Like Structures With Isolated Calf Brain Microtubule Proteins., Richard C. Weisenberg, Robert D. Allen, Shinya Inoue

Dartmouth Scholarship

Microtubule proteins isolated from calf brain will undergo gelation-contraction in the presence of ATP. We have now examined this process by video-enhanced contrast microscopy. After ATP addition to steady-state microtubules, slow (1-5 micron/min), linear movements of particles and microtubules toward aggregation centers occur. The resulting structures resemble mitotic spindle asters. During the time when gel contraction occurs, asters move (at 1-5 micron/min) toward other nearby asters. This is accompanied by the apparent shortening of the microtubules running between the asters. This is the first example of isolated microtubules undergoing a process that has similarities to half-spindle shortening during anaphase A. …

Acetylcholinesterase In The Neonatal Rat Brain: The Effects Of Prenatal Ethanol Exposure, Linda M. Boland

Biological Sciences Theses & Dissertations

Acetylcholinesterase (AChE) activity in the neonatal rat brain was determined by histochemical localization and quantitative biochemical analysis. AChE-positive perikarya were concentrated in the neostriatum, globus pallidus, medial habenular nucleus, amygdaloid complex, various hypothalamic nuclei, gigantocellular and paragigantocellular reticular nuclei, raphe nuclei, locus coeruleus, and motor nuclei of cranial nerves III-VII and IX-XII. Some areas stained differently or not at all when compared with the adult rat brain. The effects of prenatal ethanol exposure on regional AChE activity were assessed by comparing enzyme levels in offspring exposed via maternal consumption of a liquid diet containing 35 percent ethanol derived calories. This …

Seasonal Responses Of Filtration Rates To Temperature, Oxygen Availability, And Particle Concentration Of The Freshwater Clam, Musculium Partumeium (Say), Albert J. Burky, Richard B. Benjamin, Denis G. Conover, Jennifer Detrick

Biology Faculty Publications

Seasonal filtration rates of a pond population of the freshwater pisidiid clam, Musculium par­tumeium (Say), were assessed by measuring the clearance of 2.02 µ.m latex beads from suspension and expressed in terms of FR = a(AFDW}b where FR = filtration rate (ml H20 · hr- 1 ) , AFDW = mg ash-free dry weight of a whole clam and "a" and "b" are constants. The a-values ( = FR of a 1-mg AFDW clam} vary seasonally with the highest rates corresponding to periods of maximum growth and reproduction in the spring and fall. Changes in "b" reflect the influence of …

A New Column Perfusion Technique Using Isolated Rat Adrenal Cells To Study Acth And Danazol, Diane H. Johnson

Morehead State Theses and Dissertations

A thesis presented to the faculty of the School of Sciences and Mathematics at Morehead State University in partial fulfillment of the requirements for the Degree of Master of Science in Biology by Diane H. Johnson on August 15, 1982.