Research Methods in Life Sciences Commons^™

Hypoxia And Fetal Programming Of Cardiovascular Dysfunction, Qin Xue

Loma Linda University Electronic Theses, Dissertations & Projects

Human epidemiological studies have shown a clear association of adverse intrauterine environment and an increased risk of ischemic heart disease in later adult life. Of all the stresses to which the fetus is subjected, perhaps the most important and clinically relevant is that of hypoxia. The goal of this project is to test the hypothesis that chronic hypoxia during gestation adversely affects fetal cardiovascular development and impairs cardiac function in offspring. In the first part of project, we tested the hypothesis that chronic hypoxia adversely regulates contractility of fetal pulmonary arteries and veins in sheep residing at high altitude (3,801 …

Fetal Programming By Nicotine Increases Cardiac Susceptibility To Ischemic Injury, Jennifer Charlotte Alexie Lawrence

Loma Linda University Electronic Theses, Dissertations & Projects

Fetal programming is the area of study that focuses on the prenatal origins of adult onset disorders. Previous studies have associated an adverse prenatal environment with the onset of physiologic and metabolic diseases during adulthood. Fetal malnutrition, hypoxia, and exposure to drugs - such as cocaine and nicotine - have been associated with adult disease states. Cigarette smoking is the number one cause of preventable death in the developed world. Among the many dangerous chemicals found in tobacco products is nicotine, the compound responsible for the addictive nature of tobacco use. Nicotine use during pregnancy is a known cause of …

Fetal Cocaine Exposure Causes Epigenetic Changes In The Rat Heart, Kurt D. Meyer

Loma Linda University Electronic Theses, Dissertations & Projects

Cocaine abuse continues to be prevalent in the United States and other industrialized nations, in addition to the negative health effects that cocaine abuse has on the user, a mother who uses cocaine while pregnant also exposes the developing fetus to cocaine. Although there have been many studies of the effects of cocaine on the adult heart, studies of cocaine on the fetal heart and its potential delayed pathophysiological effects on cardiac function in adult offspring are extremely limited. The studies of the present project sought to enhance the understanding of the effect of cocaine exposure on the fetal heart …

Anti-Inflammatory Effects Of Ppar-Gamma In Surgical Brain Injury (Sbi), Amy Hyong

Loma Linda University Electronic Theses, Dissertations & Projects

Introduction: Brain injury can be caused by neurosurgical procedures themselves, due to direct trauma, retractor stretch, intraoperative hemorrhage and electrocautery damage. As a result of this surgical brain injury (SBI) postoperative complications such as inflammation, brain edema, and cell death can occur in the susceptible brain areas. Cerebral inflammation is a known contributor to the pathophysiology of brain injury. Following brain injury, the release of inflammatory mediators facilitates the development of BBB breakdown, cerebral edema, oxidative stress and neuronal death, resulting in further tissue damage in the brain and poor neurological outcomes. This study evaluates whether the use of a …

Neuronal Connections From The Cervix To The Spinal Cord Before Parturition, Mary Medina Groves

Loma Linda University Electronic Theses, Dissertations & Projects

By the day before birth, the density of nerve fibers in the cervix has increased compared to earlier in pregnancy or in nonpregnant mice. Similar findings are reported in parturient women. To test the hypothesis that connections between the cervix and spinal cord are enhanced by term, the cervix of nonpregnant and time-dated pregnant mice (day 15 or 18 post-breeding) was injected with pseudorabies virus (PRV), a trans-synaptic retrograde neural tract tracer. Mice were euthanized and perfused 5 days later. The spinal cord and cervix were removed and postfixed overnight, immersed in sucrose, and cryostat sections through the thoracic to …

Maturational Changes In Cerebrovascular Thick- And Thin-Filament Regulation, Renan Joel Sandoval

Loma Linda University Electronic Theses, Dissertations & Projects

The present study focused on the main hypothesis that age-related changes in myofilament Ca²⁺ sensitivity involve simultaneous changes in the relations between cytosolic Ca²⁺ transients, myosin phosphorylation, and force development. To define the effects of late fetal and early postnatal maturation, parallel experiments were carried out using cerebral arteries from both term lamb fetuses (140-141 days of gestation) and nonpregnant adult sheep. Fetal and adult cerebral arteries were stimulated by either mechanically, electromechanically, and/or pharmacomechanically-induced contractions, and the relations of cytosolic Ca²⁺, MLC phosphorylation, and contractile tensions were measured. These measurements allowed for stimulus and age-dependent …

Levels Of Intravenous Enterococcus Faecalis That Cause Heart Colonization, Louis Zane Stromberg

Loma Linda University Electronic Theses, Dissertations & Projects

While previous studies have shown that presence of bacteria in systemic circulation can cause infective endocarditis, there is no information on the specific amount of bacteria necessary to cause this condition. The purpose of this study was to establish the minimum level of circulating bacteria that will cause colonization of damaged heart valves in rats. Fifty-two Sprague-Dawley rats were anesthetized and their heart valves were damaged using an established protocol. A clinical isolate of Enterococcus faecalis was transformed with a plasmid bearing the luxF and Chloramphenicol cassettes to label the bacteria with the light-emitting protein. After four weeks, the rats …

Radiation Effects On Dopamine-Mediated Prepulse Inhibition In The C57bl/6 Mouse, Cara Nicole Zuccarelli Miller

Loma Linda University Electronic Theses, Dissertations & Projects

This study examined the effects of proton and gamma radiation on a dopamine-dependent sensory gating mechanism: prepulse inhibition of the startle reflex. The purpose of this research was to describe potential hazards of proton radiation to the central nervous system in long-term space fight and for proton therapy patients. Results from previous research indicate that startle behavior is modified by proton and iron radiation. Forty-eight 10-½ week old, male C57BL/6 mice were used to characterize the optimal parameters for the model (prepulse duration, probe duration, and stimulus-onset-asynchrony). Three hundred and fifty two mice were used to characterize the time course …

Effects Of A Serine 364 To Proline Mutation Of The Connexin43 Protein In Transgenic Mice, Chia-Ling Nhan-Chang

Loma Linda University Electronic Theses, Dissertations & Projects

[Abstract Not Included]

The Effect Of Chronic Hypoxia On Cardiac Function In The Fetal Ovine Heart, Rodney David Rodrigo

Loma Linda University Electronic Theses, Dissertations & Projects

It has been shown that long-term high altitude hypoxia during pregnancy was associated with a decrease in cardiac contractility in fetal sheep. Given that hypoxia caused apoptotic cell death in fetal and neonatal rat cardiomyocytes, the present study tested the hypothesis that chronic hypoxia during pregnancy increases apoptosis in the fetal sheep heart. Pregnant (d 30) sheep were divided between normoxic control and chronically hypoxic (maintained at high altitude, 3,820 m, PaO₂: 60 Torr for 110 days) groups. Hearts were isolated from near-term (d 140) fetal sheep, and samples from all four chambers, right and left atria, and …

Age Related Changes In Contractile Sensitivity To Cyclic Gmp And Intracellular Calcium Compartmentalization In Ovine Arteries, Surya Margono Nauli

Loma Linda University Electronic Theses, Dissertations & Projects

The study examines the hypothesis that maturation changes the ability of cGMP to produce cerebral vasodilatation and alters the relative intracellular calcium pool sizes in ovine arteries. In ovine basilars taken from near-term fetal and young adult sheep, 8- pCPT-cGMP relaxed potassium- and serotonin-induced tone with pD2 values of 4.4±0.1 and 4.9±0.1 (fetal basilar) and 4.0±0.1 and 4.7±0.1 (adult basilar), respectively. In fura-2 loaded arteries, 8-pCPT-cGMP significantly lowered increases in cytosolic calcium concentration induced by serotonin but not potassium regardless of age. When calcium sensitivity was calculated in the same arteries as the slope of the relation between calcium concentration …

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 …

Localization Of Sodium, Potassium-Adenosine Triphosphatase In The Cerebral Cortex Of Developing Rats, Maki Ujiie

Loma Linda University Electronic Theses, Dissertations & Projects

Active transport via Na⁺, K⁺-ATPase plays an important role in establishing and maintaining proper ionic gradient across all mammalian cell membranes. A strict regulation of Na⁺, and K⁺ ion levels is especially important in excitable cells such as neurons and myocytes. Inhibition of the enzyme is associated with several pathological conditions and has serious detrimental effects on cerebral cortical cells. Yet the enzyme appears to be limited in amount and distribution in newborn rats. In this study, Na_⁺, K⁺- ATPase expression and distribution in early postnatal rats were studied …

A Cytochemical Evaluation Of Blood-Brain Barrier Sodium, Potassium- And Calcium-Adenosine Triphosphatase Polarity, Panya Steve Manoonkitiwongsa

Loma Linda University Electronic Theses, Dissertations & Projects

The blood-brain barrier (BBB) is formed by cerebral vascular endothelial cells. Brain ion and fluid homeotasis essential for proper neural functioning is due to the BBB. Sodium-potassium and calcium-activated adenosine triphosphatase (Na⁺, K⁺-ATPase and Ca²⁺-ATPase) serve as one of the main mechanisms controlling brain Na⁺, K⁺, and Ca²⁺ concentrations. The present accepted concept is that both of these enzymes are localized to the abluminal plasma membrane of endothelial cells although there have been contrary results from some studies. Because of these discrepancies, further work was needed. Various cytochemical procedures …

Evaluation Of Nitric Oxide In Respiratory Distress Syndrome, Kimberly S. Myruski

Loma Linda University Electronic Theses, Dissertations & Projects

Little is known of the effects of low concentrations of inhaled nitric oxide (NO) on pulmonary mechanics and lung volumes in premature newborn infants with lung disease. Cardiopulmonary function tests including blood gas analyses were obtained from eleven premature lamb models with respiratory distress syndrome to determine if compliance, airways resistance, tidal volume, functional residual capacity, and gas exchange were affected. The effect of inhaled NO on the direction of shunting through the ductus arteriosus was determined using echocardiography.

Twin lambs were cannulated with arterial and venous catheters and intubated with an endotracheal tube before delivery on gestational day 126-130 …

Stab Wounds To Rat Brains : Demyelination, Remyelination And The Cellular Response, Particularly Of Oligodendrocytes, Xie Dangci

Loma Linda University Electronic Theses, Dissertations & Projects

The present body of knowledge concerning the results of central nervous system injury is incomplete and controversial. The role of oligodendrocytes, the cells involved in phagocytosis, the stages of demyelination, and the evidence for remyelination are of interest. Following stab wounds to rat brain cortex, corpus callosum and hippocampus, the animals were examined at 4,7,11,18,23,29,45,60 and 70 days postoperation (dpo). To help in the identification of cells immunocytochemical markers for astrocytes and immature oligodendrocytes were used. These were labels for glial fibrillary acidic protein (GFAP) and myelin/oligodendrocyte specific protein (MOSP) respectively. Light and electron microscopy were correlated using conventionally prepared …

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 …

Plasticity Of The Gnrh Neuronal System During Sexual Maturation Of The Male Djungarian Hamster, Kevin L. Buchanan

Loma Linda University Electronic Theses, Dissertations & Projects

Onset of puberty in male Djungarian hamsters is characterized by increased pituitary gonadotropin secretion and testes maturation, and is controlled by neurons within the brain which secrete gonadotropin-releasing hormone (GnRH). Puberty is associated with increased numbers of morphologically unipolar, but not bipolar, GnRH neurons in the medial preoptic area (MPOA) and diagonal band of Broca (DBB). To test the hypothesis that delayed sexual maturation arrests this increase, males were exposed to short days or administered melatonin. Males with delayed puberty had significantly fewer unipolar GnRH neuron numbers in the MPOA and DBB comparable to pubertal controls in long days. Unipolar …

Effect Of Age On The Structure Of Meissner Corpuscles In Forepad Digital Pads Of Mice, Roger C. Mathewson

Loma Linda University Electronic Theses, Dissertations & Projects

Meissner corpuscles in forepaw digital pads of albino mice were examined by light and electron microscopy to determine structural age-changes. After 1.5 months of age, no intraepidermal nerve fibers were seen extending from corpuscles. From young (1.5-6 months) to middle age (9-15 months), corpuscles became larger and more complex, gaining more horizontally arranged terminals, associated lamellae and connective tissue. At old age (18-26 months), corpuscles became small and lobulated, appearing disorganized. There was no loosening of the corpuscle-epidermal interface. An increase in collagenous connective tissue and basal lamina duplication occurred with advancing age. Ultrastructural age-changes consisted of disorganization of the …

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 …

Effect Of Photoperiod On Developmental Morphology And Enolase Isoenzyme Immunohistochemistry In Rat And Djungarian Hamster Superficial Pineal Glands, Chalmer D. Mcclure

Loma Linda University Electronic Theses, Dissertations & Projects

The best understood functional activity of the pineal gland is its diurnal production of melatonin in response to environmental lighting cues. Several enzymes of the melatonin pathway respond to daily photoperiod changes, for example hydroxyindole-O-methyltransferase (HIOMT) and serotonin N-acetyltransferase (SNAT). Increased levels of the glycolytic enzyme neuron-specific enolase (NSE) are thought to reflect increased physiological demands placed on neurons and neuroendocrine tissues. Homodimer non-neuronal enolase isoenzyme (NNE) is immunolocalized to cells, and the hybrid enolase (consisting of subunits from NSE and NNE) has been seen in cerebellar stellate and basket cells. Although not rate limiting, concentrations of both NSE and …

Regeneration Of Meissner Corpuscles In Diabetic Mice Following Median Nerve Crush, Bonnie J. Forrester

Loma Linda University Electronic Theses, Dissertations & Projects

The hyperglycemic metabolic condition found in diabetes mellitus leads to the development of sensory neuropathies and causes delayed and incomplete neuronal regeneration. Diabetic sensory neuropathy has been shown to contribute to the impairment of light touch and low frequency vibratory perception. It is proposed that the regeneration of receptors that detect these sensations will be delayed and incomplete in diabetic animals with uncontrolled hyperglycemia. This study addresses the effects of diabetes on the regeneration of Meissner corpuscles, distal sensory endings that perceive light touch and low frequency vibration.

Corpuscles from 60 diabetic mice, and their age-matched, non-diabetic littermates, were examined …

The Prenatal Development Of The "Synaptic" Structures In The Guinea Pig Pineal Gland, Zhongrong Luo

Loma Linda University Electronic Theses, Dissertations & Projects

Pineal "synaptic" ribbons are a heterogeneous population of organelles. In addition to "synaptic" ribbons (SR) sensu stricto, which are electron-dense rods, "synaptic" spherules (SS) and intermediate forms (IMF) are present. The SS are electron-dense spherical structures, the diameter of which is two to three times the width of the SR. The IMF appear as irregularly formed structures. The SR, SS and IMF are surrounded by electron-lucent vesicles. Their function and origin are unknown and also a knowledge of their prenatal development is lacking. In order to gain an insight into the development of the "synaptic structures, the pineal glands …

The Inhibition Of Bone Formation Occurring Under Weightlessness : The Effect Of Skeletal Unloading On Serum Levels Of A Bone-Derived Growth Factor, Jean D. Sibonga

Loma Linda University Electronic Theses, Dissertations & Projects

A deleterious effect of spaceflight on human health is the loss of bone associated with the prolonged lack of gravitational stress on the skeleton. A potential for bone loss is indicated by the hypercalciuria and decreased heel bone density determined in Skylab astronauts. Histological studies of juvenile laboratory rats in actual spaceflight and in simulation models document a suppression of bone formation with little alteration of bone resorption. The mechanism for net bone loss needs to be defined in order to develop measures for its prevention or correction.

Skeletal growth factors, which may mediate coupling and locally regulate bone volume …

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 …

A [6-14c] Glucose Autoradiographic Study Of The Rat Brainstem Under Hypercapnic And Hypocapnic Conditions: Identification Of Respiratory-Related Regions, Tobey M. Leung

Loma Linda University Electronic Theses, Dissertations & Projects

The effects of systemic hypercapnia on brainstem metabolism were studied with the [6-¹⁴C] glucose autoradiography method. Neuronal pools which are metabolically active take up more [6-¹⁴C] glucose and maybe revealed pictorially as darker areas on autoradiograms. Since a high concentration of carbon dioxide stimulates central chemoreceptors and hence enhances respiratory drive, the darker regions on the autoradiograms may represent regions concentrated with respiratory-related neurons. Two discrete regions in the ventrolateral medulla (a column near nucleus ambiguus and a column in the lateral reticular nucleus) appeared dark and were analyzed with an image processing system, which converted …

Hyperosmotic Modification Of Brain Extracellular Space And Blood-Brain Barrier In The Rat As Observed By Electron Microscopy, Susan A. Meacham

Loma Linda University Electronic Theses, Dissertations & Projects

The highly selective permeability of cerebral blood vessels, termed the blood-brain barrier, is due to endothelial tight junctions (zonulae occludentes). Hyperosmotic solutions reversibly open the blood-brain barrier. The purpose of this study is to determine the immediate effect of hyperosmotic arabinose perfusion upon the cerebrovascular endothelium, the morphology of the cerebral cortex, and the cortical extracellular space.

Twelve adult male Sprague-Dawley rats were separated into three groups of four animals each: control, intermediate, and extreme. Controls were perfused with a Tyrode's solution prewash followed by trialdehyde fixative. The intermediate group received a prewash of 740 mmol/kg arabinose followed by the …

Fetal Whole-Body Fluid, Cardiovascular And Endocrine Dynamics : Computer Analysis Of Regulatory Mechanisms, Peter Stuart Gold

Loma Linda University Electronic Theses, Dissertations & Projects

The dynamics of transcapillary fluid and protein distribution and endocrine regulation of cardiovascular function were studied in chronically instrumented fetal sheep during the third trimester. Average experimental changes in vascular pressures, blood volume, plasma hormone concentrations, hematocrit and plasma protein concentration were analyzed using mathematical modeling and parameter optimization techniques in order to determine values for the major parameters which regulate fluid, protein and endocrine dynamics and to determine endocrine dose-response relationships to changes in cardiovascular function. The hormones which were considered are arginine vasopressin (AVP), angiotensin II (AII), epinephrine (EPI) and norepinephrine (NE). Vascular disturbances which were considered include …

A Semiquantitative Analysis Of The Chorda Tympani Taste Pathway In The Rat Brain With The 2-Deoxyglucose Method, Charles L. Hardison

Loma Linda University Electronic Theses, Dissertations & Projects

This thesis is based on a semiquantitative analysis of four nuclear relay centers [nucleus tractus solitarius (NTS), parabrachial nucleus (PBN), ventroposteromedial nucleus (VPM), gustatory neocortex (GNC)] in the chorda tympani nerve pathway. Metabolic changes were examined by the 2-deoxyglucose method in which one chorda tympani nerve was chemically stimulated and its contralateral side lesioned. Coronal sections of four taste centers were prepared for autoradiography and Nissl staining. Densities of the autoradiograms were determined by means of a video based image processor. Relative differences in the optical density ratios of the stimulated versus lesioned nuclear grey matter areas to white matter …

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 …