Biology Commons^™

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 …

Geographic And Nongeographic Variation In The Genus Microtus (Mammalia: Rodentia) In West Virginia, Tina M. Savage

Theses, Dissertations and Capstones

Geographic and nongeographic variation was assessed on 12 cranial and 4 external characters in approximately 400 specimens of Microtus (M. ochrogaster, M. chrotorrhinus, M. pinetorum, M. pennsylvanicus) in West Virginia. Univariate and multivariate statistical analyses were used to examine age-related variation, sexual dimorphism, elevation and geographic variation and litter size for each species of Microtus in the state. Cranial characters were more beneficial than external in detecting variation. Examination of data for each species of Microtus studied demonstrated that values and trends in variation is a specific rather than a generic trait. Age-related variation was present in some, …

A New Model Species For Cultured Teleost Retinal Cell Studies: Light And Scanning Electron Microscopy, Lori Ann Steinfeldt

Undergraduate Honors Capstone Projects

Purpose: To examine retinal horizontal and bipolar cells cultured from goldfish (Carassius auratus), black crappie (Pomoxis nigromaculatus) and bluegill (Lepomis macrochirus) on the light and electron microscope levels; and to compare them with cells from a more established model, the wild white bass (Roccus chrysops). Methods: Retinas from all species were dissociated by methods developed with wild white bass (Dowling et al., 1985). Cell type was identified within the first 3 hr and cells were cultured for up to 2 wk in L-15 medium. At various intervals, cells were fixed in glutaraldehyde, …

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 …

Xenopus Laevis As A Model Organism, David C. Cannatella, Rafael O. De Sá

Biology Faculty Publications

Model organisms are often assumed to be representative of some more inclusive taxon of which the species is a part. This assumption leads to mistaken generalizations about the evolutionary and comparative significance of the data gathered. This paper reviews com? parative and evolutionary studies of Xenopus laevis and its relatives. Phylogenetic analysis of data from DNA sequences and morphology indicate that Xenopus is monophyletic and that Silurana is its sister group. The most basal lineages of Pipidae diverged prior to the breakup of Gondwana. The bizarre morphology of Xenopus is in part due to changes in the mode of meta? …

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 Influence Of 17-B-Estradiol And Tamoxifen On Protein Profiles From In Vitro Cultured Preimplantation Mouse Embryos, Darrin D. Demoss

Morehead State Theses and Dissertations

A thesis presented to the faculty of the Department of Biological and Environmental Sciences at Morehead 'State University in partial fulfillment of the requirements for the Degree of Master of Science in Biology by Darrin Lee DeMoss on August 16, 1991.

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 …

Microsurgical Fertilization Of Mammalian Eggs: An Assessment Of Clinical Utilization, Susan E. Lanzendorf

Theses and Dissertations in Biomedical Sciences

Microsurgical fertilization is the technique in which a spermatozoon or sperm nucleus is injected into the cytoplasm of an egg. To establish a foundation for the use of microsurgical fertilization as a means of treating infertility, this study evaluated the procedure in hamster and human eggs. Hamster sperm nuclei were microinjected into hamster eggs to determine the rate of abnormal fertilization and to ultrastructurally assess cellular damage by transmission electron microscopy. Hamster eggs were also injected with human spermatozoa obtained from fertile and infertile men to evaluate the fertilizing potential of the sperm cells. In addition, human eggs donated by …

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 …

Spermatogenesis And Histology Of The Testes Of The Caecilian, Chthonerpeton Indistinctum, Rafael O. De Sá, Nibia Berois

Biology Faculty Publications

Macroscopically, the testes of Chthonerpeton indistinctum consist of a series of oval lobes. The number and size of lobes is variable in Chthonerpeton indistinctum. Histologically, the testes are divided into structural units, locules separated by septa of connective tissue. Inside of each locule spermatogenesis takes place in cysts of germinal cells that divide synchronically. Chthonerpeton indistinctum is divided chronologically into four stages of development based on the shape of the nuclei of the spermatids and their arrangement in the cyst: (1) round, (2) kidney shaped, (3) attenuate, and (4) bouquet arrangement.

Conserved Chromosomal Positions Of Dual Domains Of The Ets Protooncogene In Cats, Mice, And Humans, Dennis K. Watson, Mary J. Mcwilliams-Smith, Christine Kozak, Roger Reeves, John Gearheart, Michael F. Nunn, William Nash, John R. Fowle Iii, Peter Duesberg, Takis S. Papas, Stephen J. O'Brien

Biology Faculty Articles

The mammalian protooncogene homologue of the avian v-ets sequence from the E26 retrovirus consists of two sequentially distinct domains located on different chromosomes. Using somatic cell hybrid panels, we have mapped the mammalian homologue of the 5' v-ets-domain to chromosome 11 (ETS1) in man, to chromosome 9 (Ets-1) in mouse, and to chromosome D1 (ETS1) in the domestic cat. The mammalian homologue of the 3' v-ets domain was similarly mapped to human chromosome 21 (ETS2), to mouse chromosome 16 (Ets-2), and to feline chromosome C2 (ETS2). …

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. …

The Ets Sequence From The Transforming Gene Of Avian Erythroblastosis Virus, E26, Has Unique Domains On Human Chromosomes 11 And 21: Both Loci Are Transcriptionally Active, Dennis K. Watson, Mary J. Mcwilliams-Smith, M. F. Nunn, Peter Duesberg, Stephen J. O'Brien, Takis S. Papas

Biology Faculty Articles

Human DNA segments homologous to the ets region from the transforming gene of avian erythroblastosis virus, E26, were molecularly cloned and shown to be closely related to the viral equivalent by hybridization and partial sequence analysis. The transforming gene of E26 has a tripartite origin with the structure ∆gag [1.2 kilobases (kb) from the viral gag gene]-myb(0.9 kb from the chicken myb gene)-ets (1.6 kb from the chicken ets gene). Human ets DNA is located on two distinct human chromosomes. The human ets-1 locus on chromosome 11 encodes a single mRNA of 6.8 kb; the second locus, …

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 …

Determination Of The Relative Response Of Mammalian Cells To Simulated Sunlight & 254 Nm Radiation, Randall Gill

Masters Theses & Specialist Projects

A comparison between germicidal (254 nm) UV and simulated sunlight (above 300 nm) radiation emitted from a solar simulator has been made using five mammalian cell lines. The cell lines used included normal and photosensitive human skin fibroblasts and one African green monkey kidney cell line. The ratio of 10% survival by exposure to 254 nm radiation divided by 10% survival by simulated sunlight exposure derived from the above comparison was used as a criterion for showing that the above cell lines are statistically significantly different in their relative response to simulated sunlight.

In Vivo Study Of The Effects Of Danazol On Cytoplasmic Receptors In The Female Rat, Deborah L. Spencer

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 Deborah L. Spencer on August 26, 1982.

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.

Partial Reconstruction Of The Microvillus Core Bundle: Characterization Of Villin As A Ca(++)-Dependent, Actin-Bundling/Depolymerizing Protein, Paul T. Matsudaira, David Burgess

Dartmouth Scholarship

The brush border, isolated from chicken intestine epithelial cells, contains the 95,000 relative molecular mass (M(r)) polypeptide, villin. This report describes the purification and characterization of villin as a Ca(++)-dependent, actin bundling/depolymerizing protein. Then 100,000 g supernatant from a Ca(++) extract of isolated brush borders is composed of three polypeptides of 95,000 (villin), 68,000 (fimbrin), and 42,000 M(r) (actin). Villin, following purification from this extract by differential ammonium sulfate precipitation and ion-exchange chromatography, was mixed with skeletal muscle F-actin. Electron microscopy of negatively stained preparations of these villin-actin mixtures showed that filament bundles were present. This viscosity, sedimentability, and ultrastructural …

Interpopulation Variation In Calcareous And Proteinaceous Shell Components In The Stream Limpet, Ferrissia Rivularis, Carl M. Way, Daniel J. Hornbach, Albert J. Burky

Biology Faculty Publications

Natural populations of the North American stream limpet, F. rivularis, were studied in upstate New York [USA], in a set of localities whose waters have a 15-fold range of dissolved Ca (4.6-67.6 mg/liter) and also range from oligotrophy to eutrophy. Shell component analyses (CaCO3, total organic C and total N) are reported as component mass-fractions (mg/g or .mu.g/g dry weight) and as values for a standard limpet shell of 35 mm aperture length (AL). More than 2-fold differences occur between populations in all 3 components, with relatively little variation occurring within each population. Expressed per standard limpet, CaCO3 values for …