Life Sciences Commons^™

Mechanisms Of Calcium Wave Initiation And Propagation In Astrocytes, Charles Emmet Stout

Loma Linda University Electronic Theses, Dissertations & Projects

Glial cells display propagated waves of cytoplasmic calcium (Ca²⁺) mobilization that spread outward from an initiating cell to involve hundreds to thousands of surrounding cells. This intercellular communication system is thought to initiate and coordinate Ca²⁺ dependent processes associated with vascular control, gut motility, uterine contraction, and synaptic plasticity. Not surprisingly, aberrant Ca signaling is thought to play a featured role in the pathophysiologies associated with dysfunction of these processes. This suggests that elucidation of the mechanism of Ca²⁺ waves will lead to rational therapeutic modalities for a variety of clinically significant entities.

Early studies revealed …

Molecular Basis Of An Inherited Epilepsy, Christoph Lossin, T. H. Rhodes, C. Vanoye, D. Wang, Alfred L. George

Christoph Lossin, Ph.D.

Mutations in SCN1A, the gene encoding the brain voltage-gated sodium channel alpha1 subunit (NaV1.1), are associated with at least two forms of epilepsy, generalized epilepsy with febrile seizures plus (GEFS+) and severe myoclonic epilepsy of infancy (SMEI). We examined the functional properties of four GEFS+ alleles and one SMEI allele using whole-cell patch-clamp analysis of heterologously expressed recombinant human SCN1A. One previously reported GEFS+ mutation (I1656M) and an additional novel allele (R1657C), both affecting residues in a voltage-sensing S4 segment, exhibited a similar depolarizing shift in the voltage dependence of activation. Additionally, R1657C showed a 50% reduction in current density …

Long Chain Fatty Acids And Pc12 Cell Death, Joel E. Ulloth

Loma Linda University Electronic Theses, Dissertations & Projects

Hypoxic-ischemic and traumatic injury to the central nervous system (CNS) are major sources of morbidity and mortality. The broad goal of this dissertation research is to gain a better understanding of the cascade of injury events that follow hypoxic-ischemic and traumatic injury and how they may contribute to the cell death. To accomplish this goal, one of the major secondary events common to both traumatic and hypoxic-ischemic injury was investigated: the prolonged hydrolysis of membrane phospholipid with the release of massive quantities of free (non-esterified) fatty acid (FFA). The central hypothesis of this dissertation research is that elevated levels of …

Impaired Fast-Spiking, Suppressed Cortical Inhibition, And Increased Susceptibility To Seizures In Mice Lacking Kv3.2 K+ Channel Proteins, David Lau, Eleazar Vega-Saenz De Miera, Diego Contreras, Alan Chow, Richard Paylor, Christopher S. Leonard, Bernardo Rudy

NYMC Faculty Publications

Voltage-gated K(+) channels of the Kv3 subfamily have unusual electrophysiological properties, including activation at very depolarized voltages (positive to -10 mV) and very fast deactivation rates, suggesting special roles in neuronal excitability. In the brain, Kv3 channels are prominently expressed in select neuronal populations, which include fast-spiking (FS) GABAergic interneurons of the neocortex, hippocampus, and caudate, as well as other high-frequency firing neurons. Although evidence points to a key role in high-frequency firing, a definitive understanding of the function of these channels has been hampered by a lack of selective pharmacological tools. We therefore generated mouse lines in which one …

The Role Of Brn-3.2 In Retinal Ganglion Cell Differentiation, Jerin Marie Wright

Loma Linda University Electronic Theses, Dissertations & Projects

Within the retina, the POU domain of transcription factors brn-3.0, brn-3.1, and brn-3.2 are present only in retinal ganglion cells. These genes are believed to be involved in establishing neural cell lineages in mammals. In this study brn-3.2 was examined by comparing the number of ganglion cells present during postnatal development in normal mice (+/+), in mice homozygous (-/-) for the brn-3.2 gene, and in adult mice with a heterozygous gene deletion (+/-) for brn-3.2. Optic nerve cross sections were imaged by electron microscopy, and axon profiles counted systematically by hand. These counts were then related to the nerve cross …

Regulation Of Connexin43 By Phosphorylation, Maithili M. Shah

Loma Linda University Electronic Theses, Dissertations & Projects

Gap junctions play a crucial role in embryogenesis, especially of the heart where connexin43 is thought to be important for its formation as well as synchronized contraction. Several reports have shown that connexin43 protein is phosphorylated on serine residues in vitro and in vivo.

Studies performed in our laboratory have demonstrated that cell-to-cell communication in cells expressing connexin43 can be controlled rapidly and reversibly by microinjection of active protein kinases or phosphatases that target serine or threonine residues. Phosphorylation by cAMP-dependent protein kinase (PKA) seems to favor channel opening, whereas the effect of protein kinase C (PKC) on channel gating …

The Acquisition And Quantitative Analysis Of Digitally Generated Fourier Transforms Resulting From The Physiognomy Of Various Tree And Shrub Species, Alicia Eldridge-Spears

Theses, Dissertations and Capstones

An optical Fourier transform represents the interference of wavefronts produced as light passes through multiple slits. Theoretically, any image containing periodic structure causes diffraction. The cellular arrangement of a plant leaf and the regularity in venation are ideal conditions for diffracting light. As light passes through columns of cells, its path is altered according to the distance and orientation present.

A photograph or slide of a leaf or the leaf itself may be used to produce a diffraction pattern by either optical or digital means. Optically, a laser beam directed through a slide is refracted by a converging lens and …

Archaeal Aminoacyl-Trna Synthesis: Unique Determinants Of A Universal Genetic Code?, Michael Ibba, A. W. Curnow, J. Bono, P. A. Rosa, C. R. Woese, D. Söll

Biology, Chemistry, and Environmental Sciences Faculty Articles and Research

No abstract provided.

Cellular Responses In Escherichia Coli To Lethal And Sublethal Doses Of Ozone, Indira Ruth Komanapalli

Loma Linda University Electronic Theses, Dissertations & Projects

Ozone is a major component of photochemical smog. High levels of this pollutant, sufficient to affect human health are found in many urban areas worldwide. Though limited studies in humans are supported by extensive findings from animal experiments, a difficulty in interpreting the results of these experiments has lead to an ambiguity on the biochemical mechanism of ozone toxicity. To elucidate the mechanism by which ozone causes cell damage and eventual cell death we conducted a comprehensive study using Escherichia coli K-12 as a model.

Studies on the comparative inactivation of bacteriophage lambda (λ), Escherichia coli, and Candida albicans …

The Role Of Fas And Fas Ligand In Apoptosis During Regression Of The Corpus Luteum, Sharon Roughton

Theses : Honours

Apoptosis, a form of physiological cell death, has been found to occur during regression of the corpus luteum (Juengel etal, 1993; Dharmarajan etal, 1994). The pathways involved in this process, however, have yet to be specified. One possible mediator of corpus luteum regression is the Fas (or AP0-1 or CD95) receptor, a transmembrane protein which induces apoptosis in the cell when ligated. In order to further confirm this hypolhesis, the present study establishes and quantitates the presence and regulation of Fas receptor and Fas ligand (Fasl) in the rat corpus luteum during pregnancy and post-partum. The animals used were sexually …

Ornithine Decarboxylase And Polyamines In Perinatal Hypoxia, Satyaseelan Packianathan

Loma Linda University Electronic Theses, Dissertations & Projects

Hypoxic and hypoxic/ischemic episodes contribute to various neurological aberrations, learning disabilities, et cetera in surviving infants. Despite major advances in understanding the pathophysiology of hypoxia, the cellular basis for hypoxic-mediated injury remains poorly understood. In addition, the role of developmentally essential enzymes vis á vis their contributions, or lack thereof, to hypoxic-mediated injury has not been adequately investigated. In the series of studies comprising this dissertation, we tested the hypothesis that acute and/or chronic hypoxia affect the activity of omithine decarboxylase (ODC; E.C. 4.1.1.17), the developmentally essential enzyme in polyamine biosynthesis, in the fetal and newborn rat brain. Fetal and …

Periodontal Repair In Dogs : Evaluation Of Rhbmp-2 Carriers, Lauralee Nygaard

Loma Linda University Electronic Theses, Dissertations & Projects

This study evaluated candidate carriers for recombinant human bone morphogenic protein-2 (rhBMP-2) in periodontal reconstructive surgery.

Canine demineralized bone matrix (DBM), bovine inorganic crystallene bone matrix (Bio-Oss), bovine derived microfibrillar collagen matrix (Helistat), poly (D,L-lactide-coglycolide) micropaiticles (PLGA), and polylactic acid granules (Drilac) were used with rhBMP-2 (20 (µg/100 µl implant volume) in routine critical size canine supraalveolar periodontal defects. Contralateral defects in six beagle dogs were randomly assigned to receive: DBM/rhBMP-2, DBM-control, Bio-Oss/rhBMP-2, Helistat/rhBMP-2, PLGA/rhBMP-2, or Drilac/rhBMP-2, all with autologous blood except for Helistat and PLGA. Animals were sacrificed eight weeks post-surgery and block sections of the defects were processed …

Short And Long Term Effects Of Proton Microbeam Irradiation : In The Lateral Geniculate Nucleus Of Felis Domesticus, Chad Sherwood Reder

Loma Linda University Electronic Theses, Dissertations & Projects

Little is known of the effects of proton irradiation on neural tissue. A 1.0 mm microbeam was used on the cat lateral geniculate nucleus (LGN) to quantify the short term (< 3 months) and long term (< 9 months) effects of radiation damage. The LGN is a large structure with well defined afferent, efferent, and receptive field properties.

Electrophysiological and histological techniques were used to examine the effects of the microbeam at 60, 40, and 16 Gy, administered as a single bolus. Recordings of light evoked responses in the LGN were obtained using microelectrodes in nine animals within 12 weeks of irradiation, and 6 animals within 36 weeks of irradiation. Receptive fields were mapped onto a tangent screen using standard receptive field techniques. Histological measures included afferent …

Thymic Peptide Modulates Glutathione Redox Cycle And Antioxidant Enzymes In Macrophages, Choon Sil Park

Loma Linda University Electronic Theses, Dissertations & Projects

The effect of a 6-kDa thymic peptide (TP) on the oxidative burst of a murine macrophage cell line J774 was determined. TP (12.5-200 μg/ml) was incubated with 5 x 10⁵ J774 cells at 37° C and 5% C0₂ for 18 h and oxidative burst was triggered by zymosan; chemiluminescence was amplified by luminol and measured in an automated luminometer. TP exhibited a concentration-dependent inhibition of oxidative burst. To study the mechanisms involved in TP’s inhibition of oxidative burst, its effect on the glutathione (GSH) redox cycle, GSH biosynthesis, and antioxidant enzymes was investigated. TP was shown to increase …

Transforming Growth Factor-Beta 1 Expression In Developing Ovine Teeth As Visualized By In Situ Hybridization, Søren Jepsen

Loma Linda University Electronic Theses, Dissertations & Projects

Recently we demonstrated that human dentin contained relatively high amounts of transforming growth factor beta (TGF-ß) and proposed that dentin TGF-ß originated from odontoblasts. Because of the suggested role of TGF-ß in mediating cellular interactions in dental cell development by paracrine mechanisms, we studied the expression of TGF-ß message in developing teeth by in situ hybridization. 5 µm serial sections of neonatal sheep mandibular third molars that had been fixed in glutaraldehyde and paraffin-embedded were used for in situ analysis. An [³⁵S]-labeled cRNA probe, complementary to TGF-ß1 mRNA, was constructed from human TGF-ß1 cDNA. Northern analysis of total …

Action Of Heptaminol Hydrochloride On Contractile Properties In Frog Isolated Twitch Muscle Fibre, Bruno Allard, Vincent Jacquemont, Fouad Lemtiri-Chlieh, Bernard Pourrias, Oger Rougier

fouad Lemtiri-Chlieh

1 Heptaminol stopped or delayed the progressive decline in tension which characterizes the phenomenon of fatigue in frog isolated twitch muscle fibre. 2 Heptaminol had no action on the sodium, potassium and calcium voltage-dependent ionic conductances. 3 The hypothesis of an action via an internal alkalinization was tested by comparison with the action of NH4Cl. Both substances increased the tension. 4 The action of hepataminol was suppressed in sodium-free (TRIS) solution or in the presence of amiloride while the action of NH4Cl was always observed. 5 These results could be explained by a stimulation of the Na/H antiport by heptaminol.

Alternative Temporal Control Systems For Hypodermal Cell Differentiation In Caenorhabditis Elegans, Zhongchi Liu, Victor Ambros

Victor R. Ambros

Beginning of article: In certain multicellular organisms, genetic regulatory systems that specify the timing of cell division, differentiation and morpho-genesis must accommodate environmental and physiological contingencies that perturb or arrest development. For example, Caenorhabditis elegans can either develop continuously through four larval stages (L1–L4) or arrest indefinitely as a 'dauer larva' at the second larval (L2) moult, and later resume L3 and L4 development. At the larva-to-adult (L4) moult of both con-tinuous and 'post-dauer' development, hypodermal cells switch (the 'L/A switch') from a proliferating state to the terminally differentiated state. Four temporal regulators, lin-4, lin-14, lin-28 and lin-29, have been …

Topographic Development Of The Corticocollicular Projection In The Neonatal Rat, F. Allen Richburg Ii

Loma Linda University Electronic Theses, Dissertations & Projects

The density and distribution of labeled cortical cells was examined following discrete deposits of retrograde tracers into the superior colliculus (SC) of neonatal and adult rats to study the topographic order of SC afferents from the cortex during the initial phase of axon ingrowth. Small deposits of red and green latex microspheres (0.02-0.05ul) were placed into the SC of rats ranging in age from birth to adulthood using glass micropipettes (tip diameter 25-35um). The animals, following a 48hr survival period, were deeply anesthetized and perfused with normal saline followed by a 10% formal-saline solution. The brains were sectioned coronally at …

Laminin Potentiates Differentiation Of Pcc4uva Embryonal Carcinoma Into Neurons, T. M. Sweeney, Roy C. Ogle, C. D. Little

Medical Diagnostics & Translational Sciences Faculty Publications

The embryonal carcinoma PCC4uva differentiates into neurons in response to treatment with retinoic acid and dbcAMP. We used this in vitro model system to study the effects of laminin on early neural differentiation. Laminin substrata markedly potentiate neural differentiation of retinoic acid and dbcAMP-treated cultures. Only laminin induced more rapid neural cell body clustering, neurite growth and neurite fasciculation as compared to type IV collagen, type I collagen, and fibronectin substrata. Exogenous laminin substrata promoted greater cell attachment, cellular spreading and growth to confluence than type IV collagen, type I collagen, fibronectin and glass substrata. Laminin-induced effects were inhibited by …

Characterization Of Hsd::Mudx128 Operon Fusion Mutant Of Escherichia Coli K-12, Marjorie Ann T. Reyno

Loma Linda University Electronic Theses, Dissertations & Projects

The study of the regulation of gene expression in complex genes can be facilitated by the use of operon fusions which place a well characterized lacZ gene under the control of a promoter of interest. An hsd::MudX128 operon fusion mutant of E. coli K-12 isolated by Prakash (1986) was observed to have a high β-galactosidase activity (1000 units), ten to twenty times higher than other hsd::MudX mutants. This β-galactosidase activity was suppressed to a very low level (25 units) by the introduction of an F’ plasmid, F'101, which carries the 98 min to 2 min region of the …

Presence Of A Cs-Resistant Transient Outward Current Which Distorts Ica Measurements In Guinea-Pig Single Ventricular Cells, Georges Christé, Brigitte Delachapelle, Fouad Lemtiri-Chlieh, Carlos Ojeda

fouad Lemtiri-Chlieh

To correctlyr analyse the physical properties of ion channels and the actions of drugs upon them, it is important that the membrane conducts only the ionic species believed permeant. In guinea-pig ventricular cells it has been assumed that only ICaL flows in the potential range -50 to +60 mV in the presence of intra- and extracellular Cs (> 20 mM). We report here that this is not the case due to the presence of a Cs-insensitive, transient outward current.

Measurements Of Cytosolic Free Ca In Cultured Rat Neonate And Adult Guinea-Pig Ventricular Cardiac Myocytes, Jean-Claude Bernengo, Fouad Lemtiri-Chlieh, Carlos Ojeda, Nadine Pltonoff

fouad Lemtiri-Chlieh

Cytosolic free Ca was measured with Indo-1 (Grynkiewicz et al. 1985) as free acid (in guinea-pig cells) or as the ester (in rat neonate cells). Using image analysis we obtained concentration maps at 40 ms intervals at two wavelengths (Cannell et al. 1986). In rat neonate cells, at rest and during contraction, calcium was apparently non-uniformly distributed, in contrast to guinea-pig adult cells. Using a dual PM spectroscopic system adapted to an inverted microscope we found: (1) the conversion of Indo-AM to Indo- 1 in the cytosol is not complete (< 40% contribution to the total light); (2) rat neonate cells buffer Ca (at rest 130-170 nM) only for [Ca]. between 1 and 3 mM, whereas in adult guinea-pig cells the range extended to 10 mM; (3) during contraction [Ca]i rose from 200 nm to no more than 1 microM. Digitoxin increases basal and peak [Ca]i; (4) during spontaneous contractions, in guinea-pig cells, [Ca]i rises from 70 to 200 nm when [Ca]o = 0, and from 170 nm to about 1 microM when [Ca]o = 2 mM; (5) in voltage-clamp conditions the rise in [Ca]i does not exceed 2 microM and oscillations occur in the presence of forskolin. When loading with Indo-1, cells frequently cease to contract (Powell et al. 1988) and the variations in [Ca]i are then very small (about 200 nm at peak ICa). In both cases these values are small compared to the total charge crossing the membrane.

Embryonic Chicken Fibroblast Collagen Binding Proteins: Distribution, Role In Substratum Adhesion, And Relationship To Integrins, Roy C. Ogle, A. Jeannette Potts, Marchall Yacoe, Charles D. Little

Medical Diagnostics & Translational Sciences Faculty Publications

Collagen binding proteins (CBP) are hydrophobic, cell surface polypeptides, isolated by collagen affinity chromatography. Antibodies to CBPs inhibit the attachment of embryonic chicken heart fibroblasts to native type I collagen fibrils in a dose-dependent manner. The CBP antibodies also induce rounding and detachment of cells adherent to a planar substratum. This process of antibody-mediated substratum detachment resulted in a clustering of CBP and cell-associated extracellular matrix at the cell surface, and the rearrangement of filamentous actin. Other functional studies showed that cells grown within a three-dimensional gel of type I collagen cannot be immunostained at the cell surface with CBP …

Enhancement Of Intratumor Antibody Distribution With A Bifunctional Antibody Delivery System And With Hyperthermia, Karen Lynn King

Loma Linda University Electronic Theses, Dissertations & Projects

The intratumor distribution of a new monoclonal antibody-hapten-based delivery system was examined. From two marine antibodies, Hybritech, Inc. has developed a bifunctional antibody (BPA) with specificities against CEA (ZCE-025 or GEM 231) and against one of two derivatives of [¹¹¹In]-benzyl-EDTA (CHA 255), EOTUBE or BLEDTA IV, to form ZCE/CHA or CEM/CHA. BFA accumulates in CEA-expressing tissue and clears from non-antigenic tissues prior to administration of an [¹¹¹In]-hapten.

Using five groups of nude mice bearing human colon tumor xenografts (T380), we evaluated different carriers of [¹¹¹In]-EOTUBE to prelocalized ZCE/CHA. We administered all antibodies and haptens …

Age-Related Changes In Meissner Corpuscles Of Diabetic Mice As Observed By Electron Microscopy, Vicky R. Ras

Loma Linda University Electronic Theses, Dissertations & Projects

Light touch and low-frequency vibration sense are often impaired in diabetic patients. Meissner corpuscles are distally located mechanoreceptors that respond to these stimuli. Peripheral sensory neuropathy in diabetes progresses from distal to proximal, therefore, Meissner corpuscles are ideally suited to study diabetic neuropathy in the C57BL/Ks (db/db) mouse.

Corpuscles from the forepaw digital pads of 9 diabetic and 9 nondiabetic littermates were studied by electron microscopy. These were divided into 3 age groups; 2.5 months, 10 months, and 17 months old.

Ultrastructural changes observed within various receptor components were myelin figures, multivesicular bodies, vacuoles, basal lamina duplication, abnormal and enlarged …

An Analysis Of Protein-Ligand Interactions For Enzyme Ii-Mannitol From Escherichia Coli, Brent Lee Wood

Loma Linda University Electronic Theses, Dissertations & Projects

Enzyme II^mtl of Escherichia coli was purified from mtlA-overexpressing E. coli strain LJ1112 extraction of isolated membranes with deoxycholate, followed by hexyl agarose and ω-amino hexyl agarose chromatography in Lubrol PX. A 10-fold increase in total pure protein and 2.5-fold increase in specific activity over previously reported procedures was obtained. Tryptic fragments of Enzyme II^mtl were also purified and identified as the membrane and cytoplasmic domains by protein sequencing and SDS-PAGE. The fluorescence spectrum of Enzyme II^mtl suggested that two populations of tryptophan residues existed with emission maxima at 322 and 344 nm, corresponding to buried …

Age-Related Changes Of Neurites In Meissner Corpuscles Of Diabetic, Vicky R. Ras

Loma Linda University Electronic Theses, Dissertations & Projects

This thesis is based on a qualitative and quantitative study of the neural component of Meissner corpuscles of diabetic and non-diabetic mice. Age-related changes were also documented in these mechanoreceptors. Corpuscles from forty-six diabetic mice and forty-six non-diabetic mice ranging from 2.5 to 17 months old were studied. Forepaw index digital pads were formalin-fixed and frozen-sectioned parallel to each digit and perpendicular to the skin. Serial sections were then silver-impregnated to allow light microscopic examination of the neurites in the corpuscles.

Neurites became more coarse, tortuous, branched, varicose, and thus more complex until middle age. Corpuscle size appeared greatest at …

A Light And Electron Microscopic Study Of Visual Neurons In The Medulla Externa Of The Crab Hemigrapsus Nudus, Yu Cheng Liao

Loma Linda University Electronic Theses, Dissertations & Projects

The decapod crustacean with a relatively well-defined visual system have contributed significantly to our understanding of visual neuron morphology and physiology. The general morphology of the optic lobes of several decapods were initially described by Parker (1897) and Hanstrom (1924). In the past few years, several light and electron microscope studies have been made on the lamina ganglionaris: the first optic ganglion, of the lobster (Hamori and Horridge, 1966a); the crayfish (Hafner, 1974; Nassel, 1976a), and the crab (Stowe et al., 1977; Rafuse et al., in preparation). As yet, no adequate description has been made of the visual neurons in …

The Effect Of Ergosterol On The Desaturation Of 14c-Labeled Oleic Acid In Tetrahymena, Maynard E. Neville Jr.

Masters Theses

The ciliated protozoan Tetrahymena pyriformis-Strain W desaturated ¹⁴C-labeled fatty acid substrate in both sterol supplemented (ergosterol) and nonsupplemented cultures. Cis-octadecenoic acid (¹⁴C-9-18:1), when added to the medium during the log phase of growth, was incorporated into the cells and also desaturated at the Δ⁶ and Δ¹² positions. The ¹⁴C labeled substrate and products, linoleate (9,12-18:2) or linolenate (6,9,12-18:3) were recovered and separated by silver-nitrate Unisil column chromatography. Initially, recovery of ¹⁴C compounds varied widely when expressed as the percent of total counts per minute recovered from column chromotography, and a variety of …

Cytotypes And Morphometrics Of Two Phyllostomatid Bats, Micronycteris Hirsuta And Vampyressa Pusilla, Robert J. Baker, Hugh H. Genoways, William J. Bleier, Jerry W. Warner

University of Nebraska State Museum: Mammalogy Papers

As pointed out by Jackson (1971) and Patton (1972), the key to understanding mechanisms of chromosomal evolution lies in studies of chromosomal variation within populations that can hybridize. Hybrids between cytotypes contain the answer to problems of meiotic pairing and reduced fertility, as well as information concerning the nature of zones of contact. However, before detailed studies can be initiated, chromosomal races and hybrid zones must be located. Few chromosomal races have been described for bats (Baker, 1970a). Within the family Vespertilionidae, five species are known to have such races (Baker, 1970a) and races have been reported in two species …