Research Methods in Life Sciences Commons^™

Alternatives In The 90'S: What's Next?, Henry Spira

Conference Presentations

The 80's was a decade of remarkable and innovative research and development of alternatives. We see the 90’s as the decade of validation and implementation.

From an activist's perspective, the 80's was very encouraging,-- alternatives gained acceptance, legitimacy and credibility within the toxicology, corporate and regulatory communities. And The Center for Alternatives to Animal Testing (CAAT) at Johns Hopkins University played a significant role in making this acceptance possible.

But these new technologies have yet to realize their full potential; the focus to date has been on development more than on bringing these technologies into routine usage. And the public, …

Development Of Periapical Lesions In Immunosuppressed Rats, Phillip A. Waterman Jr.

Loma Linda University Electronic Theses, Dissertations & Projects

The pathogenesis of periapical lesions is the interaction of the oral flora with the existing host defenses. The role of bacteria has been established in endodontic disease but the contribution of the host defenses in periapical lesion formation is less clear. The purpose of this study was to compare periapical lesion development in immunosuppressed rats with that in normal rats. Thirty-five, eight-week old Sprague-Dawley rats were equally divided into seven groups. Groups 1, 2, and 3 were given weekly injections of Cytoxan, an immunosuppressive agent. Groups 4, 5, and 6 received no immunosuppressive therapy. Group 7 served as a control, …

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 …

Modulation Of Rat Hepatic S9-Dependent Mutagenesis, Dna Binding, And Metabolism Of Aflatoxin B1 And Benzo[A]Pyrene By Four Chinese Medicinal Herbs, Brian Yuen Yau Wong

Loma Linda University Electronic Theses, Dissertations & Projects

Extracts of the herbs Oldenlandia diffusa (OD) and Scutellaria barbata (SB) have been used in traditional Chinese medicine for treating liver, lung and rectal tumors while extracts of Astragalus membranaceus (AM) and Ligustrum lucidum (LL) are often used as adjuncts in cancer therapy. In this study, we determined the effects of aqueous extracts of these four herbs on aflatoxin B₁ (AFB₁) benzo[a]pyrene (BaP), benzo[a]pyrene 7,8-dihydrodiol (BaP 7,8-DHD) and benzo[a]pyrene 7,8-dihydrodiol-9,10-epoxide (BPDE)-induced mutagenesis, mediated by enzymes in rat hepatic 9000 x g supernatant (S9), in Salmonella typhimurium TA100. We also studied the effects of these four herbs on …

New In Vivo And In Vitro Techniques For Quantification Of Lateral Force Levels That Cause Implants To Fail : A Progress Report On Monkey Research, V. Leroy Leggitt

Loma Linda University Electronic Theses, Dissertations & Projects

Two new techniques for quantification of forces generated by an expansion screw apparatus placed between pairs of cylindrical endosseous titanium implants are described, along with preliminary data gained from their use in Macaca rhesus monkeys. Strain gages were bonded to the expansion apparatus and were calibrated either directly by micro load cell, or indirectly by Instron Machine to reflect the applied force.

Immediate loads of up to 6 kilograms have been applied to the implants with no increase in mobility as verified by a Periotest device. No previous studies on the orthodontic uses of implants have evaluated in vivo forces …

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 …

The Ethical Judgment Of Animal Research, Shelley L. Galvin, Harold A. Herzog

Experimentation Collection

One hundred sixty subjects acted as members of a hypothetical Institutional Animal Care and Use Committee and evaluated five proposals in which animals were to be used for research or educational purposes. They were asked to approve or reject the proposals and to indicate what factors were important in reaching their ethical decisions. Gender and differences in personal moral philosophy were related to approval decisions. The reasons given for the decisions fell into three main categories: metacognitive statements, factors related to the animal, and factors related to the design of the experiment.

The Creation Of Transgenic Animal “Models” For Human Genetic Disease, Bernard Rollin

Biomedicine and Animal Models in Research Collection

Transgenic animals will be created to study human genetic disease as soon as the technological capability exists to do so. Extant laws permit such animals to be created. The mindset of the research community makes it inevitable. It is also clear that such diseases can cause enormous amounts of pain and suffering. Responsible researchers need to explore all possible avenues for controlling such pain and suffering. Thus far the research community has not engaged this issue vis a vis animals. The development of methodologies for controlling pain and suffering is likely to be exportable to numerous areas of animal research, …

Arkansas Cooperative Beef Bull Performance Test 1991, M. W. Eaton, A. E. Brown Jr., J. A. Hornsby, R. W. Parham, J. D. Shockey, W. C. Loe

Arkansas Agricultural Experiment Station Research Series

Performance testing of beef bulls was initiated to improve productivity of Arkansas beef cattle through breeding. This cooperative testing program provides cattlemen with reliable information for identifying animals with superior breeding value for the traits of production measured in the test. By using the information as a basis for herd sire selection, a breeder can greatly increase the chances of obtaining a bull that will sire rapid-gaining, more efficient, highquality calves. Such calves can increase profits for both the breeder and the feeder. This testing program provides information that is useful to financial institutes in arranging their lending programs and …

Use Morality As Basis For Animal Treatment, Kenneth J. Shapiro

Experimentation Collection

No abstract provided.

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 …

The Role Of T Cells In The Pathogenesis Of Periapical Lesions, John B. Wallstrom

Loma Linda University Electronic Theses, Dissertations & Projects

The pulps of mandibular molars of 15 athymic and 15 conventional rats were surgically exposed and left open to their oral flora. Each group was divided into 3 subgroups of 5 animals each. The rats were sacrificed after their pulps were exposed for 2, 4, or 8 weeks. After fixing, decalcifying, and embedding, the specimens were sectioned and stained with H and E. They were then examined under a microscopic grid and quantified by percentages of surface areas of bone, connective tissue, bone marrow, intrabony spaces, periapical lesions, and numbers of osteoclasts. This was done using a DataVoice computerized data …

Histomorphometric Analysis Of Primate Periodontal Tissues Following Corticotomy In Orthodontics, Wuiteng Koh

Loma Linda University Electronic Theses, Dissertations & Projects

The purpose of this study was to evaluate the prevalence of specific cell types and structures at the active tooth movement site in corticotomized and non-corticotomized specimens obtained from a previous study, and to consider the hypothesis that corticotomy facilitated orthodontics is efficacious due to an inflammatory response rather than tooth-bone block movement. Specimens from 3 monkeys, each of which had a treatment and control quadrant in each arch, were sectioned, mounted on glass slides, and stained with Giemsa, haematoxylin, and Safranin stains. A histomorphometric analysis was performed with DataVoice™, a voice activated computer system which allowed for digitization of …

Catecholamines And The Regulation Of Uterine Contractions In The Pregnant Rhesus Macaque, Cahleen Mae Mcnutt

Loma Linda University Electronic Theses, Dissertations & Projects

This study was designed to test the hypothesis that: 1) maternal and/or amniotic fluid catecholamines or prostaglandins display nocturnal activity rhythms similar to those of the myometrial contractility in the pregnant rhesus macaque and, 2) catecholamines stimulate prostaglandin production by fetal membranes. Paired maternal arterial blood and amniotic fluid samples were collected at 0900 h (AM) and 2100 h (PM) from chronically catheterized rhesus macaques until delivery and analyzed for prostaglandin metabolites (PGFM and PGEM-II) and catecholamines (norepinephrine, epinephrine, and dopamine). Uterine activity data was continuously recorded. A rhythm in uterine contractility was observed, with peak activity between 1900 and …

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 …

Animals In Biomedical Research: The Undermining Effect Of The Rhetoric Of The Besieged, John P. Gluck, Steven R. Kubacki

Experimentation Collection

It is correctly asserted that the intensity of the current debate over the use of animals in biomedical research is unprecedented. The extent of expressed animosity and distrust has stunned many researchers. In response, researchers have tended to take a strategic defensive posture, which involves the assertation of several abstract positions that serve to obstruct resolution of the debate. Those abstractions include the notions that the animal protection movement is trivial and purely anti-intellectual in scope, that all science is good (and some especially so), and the belief that an ethical consensus can never really be reached between the parties.

Reflective Ethology, Applied Philosophy, And The Moral Status Of Animals, Marc Bekoff, Dale Jamieson

Experimental Research and Animal Welfare Collection

Currently there is an unprecedented interest in ethological studies of nonhuman animals. Much of this interest is motivated by a desire to learn more about animals themselves. For scientists assuming this stance, a secondary goal is to use this knowledge to assess the place of humans in the natural order of things, stressing continuity or discontinuity depending on one's views. Others, however, study animals primarily to apply this knowledge to human behavior. We argue that behavioral research demands the rigorous application of methods that are minimally harmful to the animals being studied. We argue for a moderate, but rigorous and …

Proto-Oncogene Regulation By Growth Factors In Bone Cells, Harold Lyndon Merriman

Loma Linda University Electronic Theses, Dissertations & Projects

Previous studies have shown that bone cells in culture produce a number of growth factors that are important in bone regulation. Chick and mouse primary calvarial cultures consist of a mixture of cell types that make it difficult to interpret the results from these model systems. In contrast, MC3T3-E1 cells are a clonal mouse osteoblast-like cell line. Since MC3T3-E1 cells consist of a single population of calvarial cells, they make an ideal system in which to study the autocrine effects of bone growth factors.

Based on this work, MC3T3-E1 cells are now known to produce IGF-I, TGF-beta and IGF-II in …

Effect Of Glucose Administration On Hamster Liver S9-Mediated Mutagenesis, Metabolism And Dna-Binding Of Benzo[A]Pyrene And Aflatoxin B1, Daniel C. Rubano

Loma Linda University Electronic Theses, Dissertations & Projects

Hamster liver S9 prepared from control animals and animals given 30% glucose in drinking water 48 h before time of sacrifice was used in studies of benzo[a]pyrene (BaP) and aflatoxin (AFB₁)-induced mutagenesis, metabolism of BaP and AFB₁, and metabolite binding to calf thymus DNA. BaP-induced mutagenesis in Salmonella typhimurium TA100 was reduced 38.5% while AFB₁-induced mutagenesis was increased 36% by S9 from glucose-treated hamsters. The reduction of [³H]BaP metabolite binding to calf thymus DNA in incubations with S9 from glucose-treated hamsters correlated with a decrease in unknown BaP metabolite-deoxyribonucleoside adducts isolated by …

Understanding Dogs Through Kinesthetic Empathy, Social Construction, And History, Kenneth J. Shapiro

Sentience Collection

The term, "come into animal presence," she takes from the title of a Denise Levertov poem. The poem, which reads, in part, "What joy when the insouciant armadillo glances at us and doesn't quicken his trotting across the track into the palm bush. What is this joy?" This joy is the possibility of our being in the presence of animals for "(t)he armadillo has some intention to pursue in the palm forest." This joy, to which I invite you here, consists in dwelling in that presence, in inhabiting that intention, that armored but guileless world of the armadillo. I will …

Alternatives To And Reduction Of Animal Use In Biomedical Research, Education And Testing, John M. Frazier, Alan M. Goldberg

Experimentation Collection

Biomedical endeavours can be divided into three major categories: research, education, and testing. Within the context of each of these categories, activities involving whole animals have made major contributions and will continue to do so in the future. However, with technological developments in the areas of biotechnology and computers, new methods are already reducing the use of whole animals in certain areas. This article discusses the general tissues of alternatives and then focuses on the development of new approaches to toxicity testing.

The Effect Of An Anti-Osteoclastic Agent And A Collagen Sponge On Root Resorption Following Regenerative Periodontal Surgery In The Dog, Rudolf Hahn

Loma Linda University Electronic Theses, Dissertations & Projects

Previous studies indicate that root resorption is a frequent sequela to significant amounts of periodontal repair following experimental regenerative periodontal surgical procedures in animal models. To study if root resorption can be reduced or prevented, periodontal repair was evaluated in large horizontal circumferential defects in the dog in the absence of influence from the oral environment. Diphosphonate (Cl₂ MDP) applied topically to the root surfaces prior to wound closure as well as the use of a collagen sponge membrane (Helistat®) draped over the root surfaces and a control with no additional treatment after defect creation were the modalities employed. …

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 …

In Vitro Evaluation Of Cancer Patient Immune Responses Following Infusion Of Radiolabeled Murine Monoclonal Antibody, Kelly A. King

Loma Linda University Electronic Theses, Dissertations & Projects

Several monoclonal antibody (MAb) types derived from mice have been developed specifically for the use of targeting human tumors. The specificity of these murine MAb’s for their respective tumor antigens is very high making them potentially good immunotherapeutic agents for cancer treatment and diagnosis. A limiting factor in using these murine MAb’s in patients is the development of human anti-mouse antibody (HAMA). The presence of HAMA may reduce the effectiveness of MAb for tumor targeting. If patient sensitization could be quickly detected following the initial infusion of MAb, then future infusion could be altered or terminated to prevent further sensitization. …

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 …

Autonomic Nervous System And Cholesterol Transport, Yuan-Line Hung

Loma Linda University Electronic Theses, Dissertations & Projects

The effect of autonomic nervous system on cholesterol transport is of clinical interest because the relationship between lipoprotein metabolism and atherogenesis is expressed in coronary artery disease.

Generally speaking, beta blockers increase serum VLDL level but decrease both HDL and HDL₂ cholesterol levels without affecting the LDL in humans. We are unaware of reports concerning the effects of metoprolol on lipoprotein metabolism nor comparison of the effects of metoprolol and propranolol in the unhandled rat model. In part I we employed the unhandled rats to compared the effects of these 2 drugs administered with the drinking water in order …

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 …

Isolation And Partial Characterization Of Basic, Sperm-Specific, Nuclear Proteins In The Dog And Hampster, Richard William Hall

Biological Sciences Theses & Dissertations

The purpose was to determine whether dogs and hamsters contain the same sperm-specific protamine and testis specific proteins (TP) as other eutherian mammals previously studied. The proteins (protamine and TPs) were isolated from spermatids and mature spermatozoa, and were subjected to electrophoresis in an acid-urea, polyacrylamide gel system according to the procedure of Panyim and Chalkley (1962). The results indicated that, regardless of the anatomical location of sperm acquisition, acid precipitation technique, or denaturing agents used, epididymal and testicular samples from both the dog and hamster contained protamine in elongated spermatids and mature spermatozoa. The protamines from dogs and hamsters …

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 …