Research Methods in Life Sciences Commons^™

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 …

Mechanisms Of T Helper Cell Modulation In A Simulated Space Radiation Environment, Asma Rizvi

Loma Linda University Electronic Theses, Dissertations & Projects

Exposure to solar particle events (SPEs) poses the most serious threat to astronaut health. Due to this radiation environment, exploration of the solar system beyond Earth’s orbit entails many risks for the crew on these deep space missions. Depression or dysfunction of the immune system is a major concern due to its high radio-sensitivity; potential consequences include overwhelming infection, development of cancer, fibrosis, and poor wound healing. Astronauts are also exposed to low dose/low dose rate (LDR) radiation. A three month stay at the international space station exposes them to ~ 0.05 Gy of radiation.

C57BL/6 mice received LDR gamma-radiation …

Radiation-Induced Increases In Pkc Modulate Integrin Expression And Contribute To Fibrotic Changes, Pinal Rushikesh Pandya

Loma Linda University Electronic Theses, Dissertations & Projects

The sequences of events in fibrosis are similar to those in wound healing; however, the normal termination and resolution stages do not take place. The initial cellular response following ionizing radiation involves accumulation of the ECM (extracellular matrix) including collagen, fibronectin and the interaction of many growth factors (cytokines) with their receptors. There are a number of unanswered questions regarding many aspects of radiation-induced fibrosis, including the initial triggers and physical changes that initiate the process. We have confirmed that elevated pKC and other cellular changes following radiation are similar to those found at sites of inflammation. This information leads …

The Effect Of Radiation And Repeated Sub-Culturing On Tgf-Β1 Signaling In Frtl-5 Cells, Cheryl G. Burrell

Loma Linda University Electronic Theses, Dissertations & Projects

From our ongoing in vitro studies using the Fisher Rat Thyroid cell line-5 (FRTL-5) we recorded accelerated growth, reduced follicularization and reduction in thyroxin release that occurred as the cells were repeatedly sub-cultured. We also recorded that these changes occurred earlier and more rapidly following radiation exposure. We determined that TGF-β1 production increased under both conditions. We hypothesized that alteration in the TGF-β1 signaling pathway contributed to the changes observed in the cellular properties of FRTL-5 cells. Our objective was to examine some of the players in the TGF-β1 signaling pathway to determine whether radiation and/or repeated subculturing promoted changes …

Enterotoxin B Subunit Lectins As Adjuvants For Improvement Of Mucosal Vaccine, Nak-Won Choi

Loma Linda University Electronic Theses, Dissertations & Projects

In comparison with whole organism vaccines, subunit vaccines may be safer for immunization but may lack sufficient immunogenicity to provide complete immunity to the pathogen. To resolve this problem, bacterial and plant enterotoxin B subunit adjuvants containing a variety of receptor-binding properties were used to enhance the immunogenicity of rotavirus subunit vaccines. Enterotoxin B subunit adjuvants were employed to enhance protection against virus infection. Pentameric cholera toxin B subunit (CTB), shiga toxin-1 B subunit (STB) and monomeric ricin toxin B subunit (RTB) molecules were genetically linked to a 90 amino acid peptide from the simian rotavirus (SA11) nonstructural protein NSP4 …

Detection Of Bacteria To Dental Tissue By Low-Light Imaging, G-Hong Robert Hsu

Loma Linda University Electronic Theses, Dissertations & Projects

Bacteria have been shown to play an important role in the development of pulp and periradicular disease. The source of the infective bacteria is most often the oral cavity. Previous studies have detected bacteria in teeth that are structurally intact that seemingly have no communication intraorally. Anachoresis has been proposed as a mechanism for localizing circulating bacteria to such teeth. In several previous bacteriologic anachoresis studies, bacteria have been found to localize in inflamed dental tissues. The criticism most commonly associated with these investigations has been the potential contribution of contaminant bacteria resulting in false positive findings.

Low-light image analysis …

Development Of A Model For Infective Endocarditis Via Intravenous Injection And Tooth Infection Of Rats Using Pathogenic Bacteria, Shahrokh Shabahang

Loma Linda University Electronic Theses, Dissertations & Projects

Historically, focal areas of infection have been linked to a number of systemic ailments. During the early part of the 20th century, patients who were suffering from inexplicable diseases and who were suspected of having infected teeth were referred to the dentist for multiple extraction of their dentition. The discovery of antibiotics provided a false sense of security that the war against microorganisms is no longer a concern for mankind. However, more recently, it has become common belief that increasing numbers of systemic diseases are linked to chronic infections. Recovery of bacteria from representative tissues has led investigators to believe …

A Novel Strategy Utilizing Co-Expression Of Murine Il-12 And Antisense Tgf-Pl Against H238 Tumor Formation, Craig A. Seheult

Loma Linda University Electronic Theses, Dissertations & Projects

Cytokines have a profound effect on immune modulation, thus playing a significant role in cancer gene therapy. A variety of human carcinomas secrete transforming growth factor-beta (TGF-β), a cytokine with potent immunosuppressive properties. Suppressing TGF-β could be a key to successfully combating malignancies, such as gliomas, mammary, and colon cancers, that over-express TGF-β. Conversely, interleukin-12 (IL-12) is a potent immunostimulatory cytokine capable of regulating T and NK cell-mediated cytotoxic responses (Trinchieri, 1994) during an anti-tumor response. Activation of the immune system with such immunostimulatory cytokines renders it inert to the suppressive effects of TGF-β. To better understand these processes our …

Effects Of Liposome-Encapsulated Tnf-Alpha And Radiation Against Colon Cancer, Dong Won Kim

Loma Linda University Electronic Theses, Dissertations & Projects

Previous studies have shown that tumor necrosis factor-α (TNF-α) can augment the antitumor effects of radiation, inhibiting tumor progression to a greater extent than either modality alone. However, nonspecific toxicities associated with TNF-α have limited its use as an adjuvant. This limitation is potentially resolved with the encapsulation of TNF-α in sterically stabilized, PEGylated liposomes (STEALTH®). Thus, the overall hypothesis of the research was that the combination of radiation with STEALTH® liposome encapsulated TNF-α (SL-TNF-α) will have greater efficacy and fewer dose-limiting side effects in the LS174T human colorectal tumor xenograft model compared to the combination of free TNF-α and …

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 …

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 …

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 …

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 …

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. …

Effects Of Corticosteroids And Of Cyclosporin A On Trypanosoma Musculi In Mice And In Culture, Philip Stephen Schuyler

Loma Linda University Electronic Theses, Dissertations & Projects

The in vivo and in vitro effects of cyclosporin A and corticosteroids on Trypanosoma musculi were examined. Mice that received continuous administration of 6 mg/liter/day of dexamethasone in drinking water and 200 mg/kg hydrocortisone acetate as a single dose 24 hrs prior to injection with T. musculi had significantly higher and longer parasitemias, than controls. Body and spleen weights and B cell responsiveness were lower in test animals and there were some deaths. Mice injected with T. musculi that received 100 mg/kg/day cyclosporin A had longer, but not higher, parasitemias than controls. Corticosteroids appear to partially inactive antibody production (i.e. …

Sprague-Dawley Rats Fed Milk Or Beef Protein : Differences In Response To 1,2-Dimethylhydrazine Carcinogenesis, Stanley W. L. Ng

Loma Linda University Electronic Theses, Dissertations & Projects

Colon cancer is responsible for a high percentage of cancer deaths in developing countries, and there is convincing epidemiological evidence that meat protein and fat in the diet increase the incidence of this form of cancer. An animal tumor model which has been used by many investigators interested in colon cancer is the 1,2-dimethylhydrazine (DMH)-rat or DMH-mouse tumor model. In previous work done in this laboratory the DMH-BALB/c mouse tumor model was used to compare the production of colon tumors in mice fed a diet using non-fat powdered milk as a source of protein with those in mice fed beef …

The Effect Of Dietary Fat Level On Tumor Development And Immunity In Balb/C Mice Fed Casein Of Wheat Gluten As Protein And Challenged With Herpes Type 2-Transformed Cells, Helen Drakou

Loma Linda University Electronic Theses, Dissertations & Projects

A high percentage of fat in the diet has been implicated through epidemiological studies in several human cancers. Studies in experimental animals have produced evidence that increasing the fat in the diet decreases the animal’s ability to restrict tumor development and growth. In previous work done in this laboratory with BALB/c mice injected with herpes simplex Type 2-transformed mouse cells (H238), the effects of a diet which greatly enhanced tumor growth was compared with one which restricted tumor growth. The latter, a low (5%) fat, low protein diet, in which the protein was casein, was also found to produce a …

A Rapid Concentration And Identification Technique For Human Malaria Infections Using A Mouse Model, Donald D. Barton

Loma Linda University Electronic Theses, Dissertations & Projects

Red blood cells infected with malaria parasites experience a change in density making them lighter (less dense). This paper reports a new, rapid diagnostic technique using this change in density in the infected red blood cell. It is not only easy to use, but it is also compatible with the technological conditions found where the malaria problem is most acute - in many developing countries of the world. A small sample of blood drawn from a simple finger stick can be concentrated and examined microscopically for malaria parasites within 10 minutes. The thick film slide preparation, presently used, takes a …

Cryptosporidiosis In The Immunosuppressed Mouse And In Cell Culture, Mercy Prabhu Das

Loma Linda University Electronic Theses, Dissertations & Projects

Cryptosporidiosis is a parasitic infection primarily of animals that has been recently recognized as an important disease in immunocompromised humans. Most cases to date have occurred in patients with acquired immunodeficiency syndrome (AIDS). Persons with abnormal humoral or cellular immune function, and even normal subjects, have also been affected.

To determine whether or not mice, when experimentally immunosuppressed, would develop cryptosporidiosis, a test was designed using the AKR/J strain. This strain was selected because, it was previously reported that it was the strain shown to respond most readily to the production of disease caused by the opportunistic protozoan, Pneumocystis carinii …

Cell Mediated Immunity By Cytotoxicity Assay And The Effect Of Corynebacterium Parvum And Radiation On Mice Bearing Herpes-Induced Tumors, Robert B. Stagg

Loma Linda University Electronic Theses, Dissertations & Projects

Cell mediated immune (CMI) cytotoxic reactivity of Balb/c mice against H238 cells, a Herpes simplex virus (HSV) Type 2 - induced sarcoma, was measured by the ¹²⁵IUdR release assay. The Balb/c mouse response to the growing HSV-induced tumor, treated with radiation and , was measured by survival rate, tumor growth and immune cytotoxicity of spleen and peritoneal exudate cells (PEC) as determined by ¹²⁵IUdR release assay. Subcutaneous (s.c.) inoculation of 1 x 10⁶ H238 cells (high dose) produced progressive tumor growth while s.c. inoculation of 1 x 10⁴ H238 cells (low dose) produced no tumors. A …

Effects Of Trypanosoma Musculi Kendall, 1906 Immune Serum On The Reproduction Of Trichinella Spiralis (Owen, 1835) Railliet, 1896 In Mice, Samuel L. Chafin

Loma Linda University Electronic Theses, Dissertations & Projects

The possible effects of ablastin, a reproduction-inhibiting antibody which is produced during the infection of the blood parasite, Trypanosoma musculi in the mouse, were investigated by fractionating the parasite into a supernatant fraction (designated as Group A test mice) and a pellet fraction (Group B test mice). These groups A and B mice were injected with the respective fractions, plus Freund’s Complete Adjuvant (FCA). Control mice were injected with FCA only. Sera from these three groups of mice were removed after the Group A and Group B mice were shown to be negative to T. musculi challenge, and Group A …

Effect Of Radiation And Corynebacterium Parvum On The In Vivo Elimination Of Friend Virus-Induced Leukemia In C57bl/6 Mice, Betty A. Wu

Loma Linda University Electronic Theses, Dissertations & Projects

Administration of Corynebacterium parvum in combination with localized irradiation has been shown to be more efficient than radiation treatment alone in prolonging the survival time of methylethrane induced tumor in C3H mice. Fractionated low dose total body irradiation has been demonstrated to be a promising therapy for patients with chronic lymphocytic leukemia. The purpose of this study was to determine the effect of the combined treatment of total body irradiation and C. parvum on Friend virus induced leukemia in C57BL/6 mice. ^₁₂₅I-labeled FBL-3 cells were injected intraperitoneally into mice. Intraperitoneal injection of C. parvum and and single dose total …

Concurrent Infection Of Trypanosoma Musculi Kendell, 1906, And Schistosoma Mansoni Sambon, 1907 In Mice, Berhane Andeberhan

Loma Linda University Electronic Theses, Dissertations & Projects

The possible effects of ablastin on Schistosoma mansoni were investigated by (1) establishing a concurrent infection of Trypanosoma musculi and Schistosoma mansoni in mice, and (2) by obtaining Trypanosoma lewisi immune rat serum (ablastin) and injecting it into S. mansoni infected mice. These procedures were followed on days 4, 14 and 28 of S. mansoni infection. For the groups that were given immune rat serum, controls were kept into which normal rat serum was injected.

Ablastin does not have any effect on the development or morphology of the S. mansoni worms as tested in this study. Although ablastin inhibits glycolytic …

Concurrent Infections Of Trypanosoma Equiperdum And Trichinella Spiralis In Mice, Pauline A. Nembhard

Loma Linda University Electronic Theses, Dissertations & Projects

In concurrent infections of Trichinella spiralis and Trypanosoma equiperdum, the survival times vary in the early stages of the Trichinella infection. On Days 1 and 4, the mean survival times are significantly increased, and on Days 3 and 7 they are markedly decreased. This decrease is probably due to intestinal tissue invasion. Mice infected with trypanosomes after the 7th day of the Trichinella infection have survival time increases to a significant point and a steady leveling off to Day 21. It is suggested that the antibodies produced against the Trichinella worms slow down multiplication of trypanosomes and increase the …

The Germfree Culture Of Three Species Of Triatominae: Triatoma Protracta (Uhler), Triatoma Rubida (Uhler), And Rhodnius Prolixus Stål, Stephen A. Nyirady

Loma Linda University Electronic Theses, Dissertations & Projects

Triatoma protracta protracta, Triatoma riibida rubIda, and two strains of Rhodnius prolixus were successfully reared germfree into the fourth generation. Triatominae eggs were disinfected externally with iodine and the nymphs reared in vinyl plastic germfree modules. Germfree mice were used as the host. This is the first report of Triatominae having been reared in a germfree environment. These bugs were never exposed to bacteria internally or externally to the fourth generation.

Comparisons of conventional control bugs with replicate germfree groups in the first two generations revealed but slight differences in developmental periods. The mean adult longevity of all …

The Effect Of Elevated Glucose On Neutrophilic Phagocytosis In Genetically Diabetic Mice, Larry B. Thompson

Loma Linda University Electronic Theses, Dissertations & Projects

Infiltration to an injury site or area of bacterial contamination by polymorphonuclear leucocytes (or neutrophils) is an early and essential phase of the early inflammatory reaction. Their action on foreign matter is the most critical step taken by body defense mechanisms in the first 24 hours. Anything which interferes with that action, phagocytosis, greatly hampers early inflammation. This study was designed to see the effect of elevated glucose levels on neutrophil phagocytosis in a unique strain of genetically diabetic mice.

Twenty-two diabetic and twenty-four control female mice were used. Blood glucose determinations and total white counts were made on each …

Infectivity And Development Of Trichinella Spiralis In Germ-Free, Monocontaminated And Conventional Mice, Daulat Sitanggang

Loma Linda University Electronic Theses, Dissertations & Projects

The purpose of the experiment was to compare the development of Trichinella spiralis in germ-free, Lactobacillus casei monocontaminated, Escherichia coli monocontaminated and conventional mice. The criteria used in the experiments were the recovery of adults and larvae of T. spiralis after 72 hours and one month post-infection, respectively. which gave an indication of the effects of no bacteria or only a certain, single species of bacterium on the development of this nematode in mice.

Germ-free T. spiralis larvae were obtained by washing them several times with sterile normal saline, and treatment with antibiotics. The gavaging procedure was done without anesthetization …

The Intestinal Flora Of Triatoma Protracta Protracta (Uhler) (Hemiptera: Reduviidae) Including New Bacterial Isolation Techniques, Stephen A. Nyirady

Loma Linda University Electronic Theses, Dissertations & Projects

A bacteriological procedure for the removal of the intestinal flora of Triatominae is described. Immersion of Triatoma protracta adults and fifth instar nymphs in 5% aqueous formaldehyde for three hours was sufficient to free the integument of these insects of viable bacteria without penetration into the alimentary tract.

Twenty-two adults and 17 fifth instar nymphs of Triatoma protracta protracta were collected from Neotoma lodges in southern California. Eleven of the 17 nymphs molted into adults in the laboratory. With the use of a 2% peracetic acid sterilized custom built plastic germ-free isolator the Triatoma were dissected and various sections of …

The Fecal Shedding Of Listeria Monocytogenes By Conventional And "Germ Free" Mice, Margaret Joyce Ramsay

Loma Linda University Electronic Theses, Dissertations & Projects

A series of experiments involving Listeria monocytogenes was undertaken to study first the fecal shedding of Listeria by conventional mice challenged intraperitoneally with Listeria and second the fecal shedding and ability to survive of germ free mice with similar challenge. In addition, mites infesting Listeria infected mice were cultured for their possible contamination by Listeria.

Conventional mice were challenged intraperitoneally with a single near LD₅₀ dose of Listeria. Fecal Listeria were detected by collecting specimens from each mouse daily for the first 4 days and every second day thereafter. The feces was held at 7°C for 14 …

The Effect Of Varying Protein Intake On Trichinella Spiralis Infection In Mice, Clare K. Kwan

Loma Linda University Electronic Theses, Dissertations & Projects

Three hundred Swiss Webster albino mice three weeks of age were placed on low (8%), medium (24%), high (48%) protein levels and a commercial laboratory animal feed, Purina Chow for various lengths of time: (1) simultaneously with infection, and four weeks thereafter, (2) three weeks before infection and throughout the four-week infection period, and (3) six weeks prior to infection and during the infection period. The protein source was casein. Sixty of the 300 mice used served as noninfected controls, and the other 240 were each infected with 400 washed, suspended Trichinella spiralis larvae obtained from a rat by the …