Laboratory and Basic Science Research Commons^™

Vaccinia Virus-Mediated Glioma Gene Therapy Using P53, Il-2 And Il-12, Bing Chen

Loma Linda University Electronic Theses, Dissertations & Projects

Malignant brain tumors are generally lethal soon after their diagnosis, and the five-year survival rate for the patient is low in spite of neurosurgery, radiation therapy, and/or chemotherapy. Gene therapy has the potential to improve patient survival and quality of life by delivering specific combinations of therapeutic genes directly to the tumor for localized treatment.

We have based our cancer gene therapy for glioma on a highly attenuated variant of the Lister strain of vaccinia virus (VV) which has been proven to be a safe and effective vaccine vector. Other advantages include a wide host range, a strong promoter system, …

Identification And Characterization Of The Cis-Acting Elements Around The Murine Cd4 Cnhancer, Xin Dong

Loma Linda University Electronic Theses, Dissertations & Projects

The cluster determinant 4 (CD4) molecule is a transmembrane glycoprotein. CD4 is essential for normal T helper cell function and plays an important role in T cell development and activation. CD4 is encoded by a single gene located on chromosome 6 in the mouse and chromosome 12 in the human. Both human and murine CD4 genes are divided into ten exons spanning more than 25 kb and have a large non-coding region in the first and third intron. CD4 gene expression is controlled primarily at the transcriptional level during T cell development and activation.

An enhancer has been identified approximately …

Connexin-32 Contributes To Radiation Resistance In Thyroid Follicles, Da-Thao Dinh Tran

Loma Linda University Electronic Theses, Dissertations & Projects

The objective of this study was to examine the role of gap junctions in the radiation resistance of thyroid follicles. The question of whether gap junctional intercellular communication plays an important role in the “contact effect”, wherein cells in direct apposition are more resistant to the damaging effects of radiation, has been debated for years. We used the Fischer rat thyroid cell line (FRTL-5) to carry out our experiments. This thesis is divided into two parts: 1) characterization of gap junctional properties in FRTL-5 cells and, 2) exposure of these cells to radiation.

We found that, unlike primary thyroid cells, …

Identification And Characterization Of Control Elements Within The Murine Cd4 Gene, Zhong Deng

Loma Linda University Electronic Theses, Dissertations & Projects

The control of CD4 gene expression is essential for T lymphocyte development. Since the molecular mechanism for the control of CD4 gene expression during T cell development had not been elucidated, a study of the factors that control CD4 gene expression may lead to further. Toward these goals, we have made a series of recombinant DNA constructs to define the cis-acting transcriptional control elements in the murine CD4 locus that control CD4 gene expression during T cell development. In this study, we have identified multiple cis-acting control elements, which are critical for regulating the expression of the murine …

Periradicular Response Of Immunodeficient Rats To Mechanical Pulpal Exposure, James E. Stich

Loma Linda University Electronic Theses, Dissertations & Projects

Periradicular lesions develop as a result of the interaction between root canal irritants and the immune system. The purpose of this study was to investigate the periradicular response of immunodeficient rats to their oral flora. Twenty Sprague-Dawley rats received a total body irradiation dose of 750 cGy one week prior to the investigation. The pulps of the mandibular first molars of these animals and six nonirradiated rats were exposed mechanically with a ½ round bur and left open to the oral cavity. Additional irradiation (350 cGy) was administered bi-weekly to maintain immunosuppression. Periradicular specimens were obtained after two and four …

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 …

The Immunology Of Parturition, Ari Mayer Mackler

Loma Linda University Electronic Theses, Dissertations & Projects

Macrophage-derived factors have been implicated in the etiology of preterm labor but little is known about trafficking of this immune cell into the uterus or its regulation of uterine contractility in mice. Enhanced number and activation of macrophages was hypothesized to precede parturition. Studies characterized contractile activity and the distribution of macrophages in the uterus of C3/HeN mice before and after term (=day 19). Contractile activity by uterine strips on days 15 and 18 of pregnancy, the day of delivery, and 1 day postpartum indicate that an endogenous oscillator sustains high frequency contractions. Modest acceleration of the pacemaker occurred before …

A Pilot Study Testing A Proprietary Sealant For Plaque Reduction, Jennifer Rowland

Loma Linda University Electronic Theses, Dissertations & Projects

Enamel demineralization due to increased plaque accumulation is a well-recognized problem associated with fixed orthodontic appliances. The purpose of this study was to evaluate the efficacy of a proprietary product to be marketed by 3M Unitek for the reduction of plaque accumulation and thereby reduce enamel demineralization.

Sixty four bovine teeth were divided equally into an untreated control group and a group treated with the proprietary sealant product. Both groups were subsequently immersed in a Streptococcus mutans culture. Plaque deposits from each group were removed, suspended in trypticase soy broth, serially diluted, and plated on trypticase soy agar at 1, …

The Effect Of Root End Demineralization On Cementogenesis : A Long Term Investigation, Hamid R. Abedi

Loma Linda University Electronic Theses, Dissertations & Projects

Craig and Harrison¹² examined the effect of demineralization of resected root ends on wound healing following periradicular surgery. Based on its microscopic evaluations from 4-45 days, it was proposed that demineralization of resected root ends enhances cementogenesis. The purpose of this study was to investigate the longer term effects of demineralization on cementogenesis using Mineral Trioxide Aggregate (MTA) or amalgam as root end filling materials. After completion of root canal therapy and endodontic surgery on seventy-two roots of mandibular second, third and fourth premolars of six beagle dogs, 32/72 of the root ends at random were etched with 50% …

Study Of The Hiv-1 Gene Regulatory Proteins Tat And Rev By The Two-Hybrid System In Yeast, David Allen Elkins

Loma Linda University Electronic Theses, Dissertations & Projects

Tat and Rev, small proteins encoded by the Human Immunodeficiency Virus type 1 (HIV-1), regulate its pattern of gene expression within infected cells. Tat increases transcription from the integrated pro virus by approximately a thousand-fold; Rev effects a shift in the pattern of splicing of the viral mRNA. Both are likely to interact with cellular proteins in executing their respective functions. Tat has additional activities (including neurotoxicity and inhibition of lymphocyte activation) which may be extraneous to its viral functions.

In order to identify cellular proteins interacting with Tat and Rev, the genes for both were cloned into plasmid vectors …

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 …

An Animal Model For Development Of Pulpal Inflammation, John W. Masters

Loma Linda University Electronic Theses, Dissertations & Projects

Several animal models have been proposed to develop pulpal inflammation. Procedures recommended in these models have many variables and have not produced consistent results. The purpose of this study was to create pulpal inflammation using the Nd:YAG laser.

Forty-five cuspids of 12-18 month old beagle dogs were lased at 5, 7.5, and 10 watts (W) for 20 seconds with a 600 micron contact probe. Five cuspids were not lased but were used as controls. Following laser application, the teeth were extracted at intervals of 3, 7, and 30 days. They were then fixed in 10% formalin, decalcified, sectioned, and stained …

In Vitro Plaque Formation On Orthodontic Resin Sealants, Leland W. Pack

Loma Linda University Electronic Theses, Dissertations & Projects

The purpose of this study was to examine in vitro plaque formation on sealant coated enamel. The plaque growth on different types of sealants placed on enamel was compared to each other and to an unsealed enamel control. The antimicrobial properties of sealants were also studied.

Sealant coated bovine enamel disks were immersed in a S. mutans culture medium for four days. The amount of bacterial plaque formation was quantified by removing the deposit, suspending it in solution with subsequent serial dilution. Aliquots of the dilutions were then spread on blood agar plates and incubated for 48 hours. The number …

The Effect Of Guidor™ And Hop-1 On Hard Tissue Healing : Following Periradicular Surgery In Cats, Hugh Maguire

Loma Linda University Electronic Theses, Dissertations & Projects

The purpose of this study was to evaluate the effects of the resorbable membrane Guidor™ and the osteogenic protein hOP-1 on hard tissue healing following periradicular surgery in cats.

Surgical endodontic therapy was performed on the maxillary canine teeth of twelve cats. After orthograde root canal therapy, flap reflection, osteotomy, resection of the root end, root end cavity preparation and the placement of a root end filling, eight of the surgical sites were treated with Guidor™ and eight with hOP-1. The remaining eight sites received no further treatment and served as controls.

The animals were sacrificed at twelve weeks following …

Bone Morphogenetic Proteins And Periodontal Repair, Michael B. Lee

Loma Linda University Electronic Theses, Dissertations & Projects

For over thirty years now, research has been carried out to isolate and purify bone morphogenetic protein (BMP), a substance which has been shown to induce heterotopic bone formation in various animal species. Recent advances in the fields of developmental biology, molecular biology and genetics, have shown that the BMPs not only are responsible for postfetal bone induction (including normal bone remodeling, healing and repair), but are also critical during embryogenesis - not only in regards to the skeletal system, but quite possibly in the morphogenesis and pattern formation of other tissues and organs as well.

Due to their bone …

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 …

The Production And Characterization Of A Monoclonal Antibody To A Naturally Occurring Aminomalonic Acid Epitope And A Comparison With An Antibody To A Synthetically Derived Aminomalonic Acid Epitope, James L. Gulley

Loma Linda University Electronic Theses, Dissertations & Projects

The goal for this project is to undertake the first step in the study of blocking metastasis by a monoclonal antibody. The long range goal of this line of research is to study a novel mechanism for the blocking of tumor metastasis in general and prostatic cancer metastasis in particular. The problem that this addresses is the growing number of cases of prostatic cancer in the increasingly aging male population. This cancer is not lethal unless it metastasizes, yet there is no cure for the metastatic disease. The inherent specificity of a monoclonal antibody combined with the uniqueness of the …

Effects Of Furosemide And Oleic Acid On Thyroxine Binding To Isolated Pairs Of Thyroxine Binding Serum Proteins, Deborah Hustead

Loma Linda University Electronic Theses, Dissertations & Projects

Dosage dependent binding inhibition and redistribution of thyroxine (T₄) between three pairs of thyroxine binding serum proteins, TBG-TTR, TBG-albumin and TTR-albumin, by furosemide (0.0 to 1.0 mol/L) and oleic acid (0.0 to 0.10 mol/L) were studied using equilibrium dialysis with purified binding proteins at concentrations 1/200 of levels in normal human sera, separated across the dialysis membrane and competing for T₄.

Furosemide (1.0 mol/L) reduced TBG bound T₄ in both the TBG-TTR system and TBG-albumin system (>90% decrease) and inhibited T₄ binding to TTR in the TTR-albumin system (68% decrease) and to albumin …

Healing Potential Of Osteotomies Of The Nasal Sinus In The Dog, Roderick W. Tataryn

Loma Linda University Electronic Theses, Dissertations & Projects

Maxillary sinus osteotomy is sometimes indicated for transantral surgical approaches to palatal roots of maxillary molars. A search of the literature showed an absence of any report on the healing potential of such defects. The purpose of this study was to determine histologically the reparative process of the nasal sinus of dogs following small and large surgical antral perforations. After anesthetizing six beagle dogs, full thickness periosteal flaps were raised overlying the maxillary buccal cortical plates. A 5x5 mm diameter perforation was created with a trephine bur above the second maxillary premolar on one side of each animal. On the …

Suppressive Effects Of Transforming Factor-Β And Interleukin-10 On The Cytolytic Activity Of Murine Macrophages And Reversal By Cytokines, Chin-Hung Lin

Loma Linda University Electronic Theses, Dissertations & Projects

In this study, the suppressive effects of transforming growth factor-β (TGF-β) and interleukin-10 (IL-10) on peritoneal macrophage killing of H238 target cells and the potential for reversal of the immunosuppressive effect by IL-4 and interferon-γ (IFN-γ) were investigated. The responsiveness of naive and peptone-activated macrophages was compared. The cytolytic activity for tumor cells of these effector cells was measured by percent lysis of ³H-thymidine labeled Herpes simplex virus type 2-transformed tumor cells (H238). After 18-24 hours of incubation with TGF-β or IL-10, the cytolytic activity of macro-medium alone. The immunosuppressive effect of TGF-β or IL-10 on non-activated macrophages was …

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, …

An Evaluation Of Surface Enamel Damage Following The Debonding Of Ceramic Brackets : A Follow-Up Study, B. Scott Hood

Loma Linda University Electronic Theses, Dissertations & Projects

To evaluate surface enamel damage after debonding ceramic brackets using their manufacturer's recommended methods, sixty four bicuspid teeth were bonded with three different types of ceramic brackets and one type of metal bracket. All the brackets tested utilized mechanical retention only. The brackets were debonded in the mouth seven to ten days after bonding, extracted and evaluated with a scanning electron microscope. The study concluded that: 1) all brackets alter the surface enamel, 2) there is a significant difference in surface enamel damage between the metal bracket and the ceramic brackets utilizing a tensile debonding method, 3) there is no …

A Flow Cytometric Analysis Of Immunocyte Populations In Peripheral Blood And Pleural Effusions Of Patients With Cancer, Mandy L. Bohn

Loma Linda University Electronic Theses, Dissertations & Projects

Two-color flow cytometric analysis was performed on paired samples of peripheral blood (PB) and pleural effusions (PE) of patients with metastatic malignancies. The purpose of these analyses was to test the hypothesis that there are significant differences in the distribution of immunocyte populations in the PE compared to that of the PB.

The majority of the pleural fluid immunocytes were CD3+ cells. The ratio of CD4+ to CD8+ cells was higher in the PE compared to PB. Levels of CD8+, CD19+ and CD14+ cells were not significantly different in the PE compared to the levels in the PB. CD16+CD56+ cells …

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 …

Molecular Mechanism Of The Stimulation Of Alkaline Phosphatase Activity In Human Bone Cells By 1,25(Oh)2 D3, Eru Kyeyune-Nyombi

Loma Linda University Electronic Theses, Dissertations & Projects

To facilitate this study an in vitro human model system was established that exhibited many aspects of normal osteoblasts. The human osteosarcoma cell line (TE85 cells) expressed a skeletal alkaline phosphatase activity (an accepted bone cell differentiation marker) which was stimulated by 1,25(OH)₂D₃ (a potent differentiating agent), under serum-free conditions in a dose-dependent, time-dependent, and cell density-dependent manner. Cytochemical analysis of the stimulation of ALP activity by 1,25(OH)₂D₃ showed that 1,25(OH)₂D₃ increased the number of TE85 cells that expressed detectable ALP activity, suggesting that 1,25(OH)₂D₃ promoted the process …

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 …

Pharmacokinetics And Effects On Blood Chemistry Of Cyclosporine In Human Infants And Kid Goats, Floyd A. Beierle

Loma Linda University Electronic Theses, Dissertations & Projects

Kid goats (newborn) have been used in transplantation protocols to study the techniques and problems of infant human cardiac transplantation. Immunosuppression has been by cyclosporine in both species. However, the pharmacokinetics of cyclosporine (CSA) in both the kid goat and human infant were unknown. Therefore, the pharmacokinetics of CSA were investigated. A single dose study and a long term chronic administration study were undertaken in the goat. Six Nubian goats, ages 60-90 days, were given a single IV dose of CSA, 0.25 mg/kg, over a 10 minute period and 72 hr later an oral cyclosporine dose of 20 mg/kg. The …

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. …