Microbiology Commons^™

Comparison Of Five Different Serological Assays For Determination Of Antibody Levels Against Herpes Virus Type 1 And 2, Murat Sami Küçük

Loma Linda University Electronic Theses, Dissertations & Projects

To aid in the diagnosis of certain clinical types of Herpes virus infections, we evaluated and compared five serological methods for measuring Herpes virus antibody. The five tests we compared are complement fixation test (CF), indirect immunofluorescent antibody assay (IFA), anticomplementary immunofluorescent assay (ACIF), enzyme linked immunoassay test (ELISA), and the latex agglutination test (LA). The main objective of this study was to establish which of the currently available serologic tests was best suited for clinical laboratory use and which may then replace the complement fixation test. In the comparative testing, one hundred twenty serum samples were divided into four …

Determination Of Antibody Levels In Periapical Lesions Against Sixteen Oral Microorganisms By The Elisa Technique, Sammee Lee Jones

Loma Linda University Electronic Theses, Dissertations & Projects

The root canal system, which includes the periapical region of the tooth, may become infected by several types of microorganisms which are normally associated with these tissues. A bacteria-specific antibody response may occur at the local level, where various classes of immunoglobulins have been found and reported in the literature. The purpose of this study was to quantitatively measure locally produced immunoglobulin' levels in periapical lesion abscess materials and to determine the specific reactivity of these antibodies to several microorganisms which are normal flora, as well as being previously associated with the development and pathogenesis of endodontic infections. Periapical lesions …

Immunologic Mechanisms Involved With Human Endodontic Lesions, Sunita V. Prabhu

Loma Linda University Electronic Theses, Dissertations & Projects

Endodontic or root canal infection results in immunologic changes in periapical region, which causes the destruction of pulpal and periapical tissue. No definitive study has identified a specific organism or a group of organisms as a etiological agents. Past studies have indicated that immunoglobulin E play a significant role in the pathogenesis of periapical lesions. The presence of this factor suggests that some of the endodontic flare-ups may be IgE-mediated hypersensitivity reactions. Antibody levels of IgA have been quantitatively measured by ELISA and IgE by modified RAST assay, against eleven oral bacteria which are routinely isolated from such lesions. The …

The Isolation And Characterization Of Growth Regulatory Factors Produced By A Herpes Simplex Virus Type 2 Transformed Mouse Tumor Cell Line, H238, Robert B. Stagg

Loma Linda University Electronic Theses, Dissertations & Projects

Transformation of cells with herpes simplex virus Type 2 (HSV- 2)occurs by an unknown mechanism. No specific gene or gene product has been consistently associated with HSV-2 transformation as is the case for other tumor viruses. This study was performed in an attempt to associate HSV-2-transformation with specific growth factors in order to develop a testable model for HSV-2-transformation. For some tumor viruses, particularly those containing RNA, there has been a central concept (autocrine secretion) linking oncogenes and growth factors. This concept centers on the ability of the cancer cells to produce and respond to their own autologous factors. We …

Chemoreceptors Regulate Discharge Of Microbasic P-Mastigophores In The Sea Anemone, Aiptasia Pallida, Gail E. Muir Giebel

Loma Linda University Electronic Theses, Dissertations & Projects

Recently, using cnida-mediated measurements of adhesive force, Thorington and Hessinger (1984, 1988a, b) identified two different chemoreceptors involved in triggering cnida discharge on the tentacles of the sea anemone, Aiptasia pallida. These two classes of receptors were shown to interact with glycine and N-acetylneuraminic acid (NANA). We now show that the discharge of the microbasic p-mastigophores, one of the three types of cnidae present on the tentacles of A. pallida, is under the controlling influence of these two classes of cnidocyte-associated chemoreceptors. We also demonstrate that the number of discharged microbasic p-mastigophores adhering to the test probes is proportional to …

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. …

Dosage Window Of Selenium On Human Natural Killer Cytotoxicity And Neutrophilmediated Chemiluminescence, Mary Labue

Loma Linda University Electronic Theses, Dissertations & Projects

Few studies have been conducted on the effect of in vitro selenium (Se) supplementation on cellular immunity. In the present study, the effects of in vitro incubation with sodium selenite on natural killer (NK)-mediated cytotoxicity and phagocyte chemiluminescence (CL) were examined. NK-mediated cytotoxicity and phagocyte CL were observed to be affected by added selenite. These changes generally included augmentation and suppression of both neutrophil CL and NK-mediated cytotoxicity. All dosages of selenite used were shown to be capable of causing augmentation of neutrophil CL. However, 1.0 /xg/ml selenite was also observed to occasionally cause depression of neutrophil CL. When Se …

The Determination Of Antibody Levels In Endodontic Patients Against Eleven Microorganisms Using Ifa And Elisa Techniques, William C. Payne

Loma Linda University Electronic Theses, Dissertations & Projects

Endodontic (root canal) infections are generally considered to be caused by the invasion of microorganisms which destroy pulp tissue and extend to the periapical region. No definitive study has identified a specific bacterium or group of bacteria as etiological agents. We quantitively measured antibodies present in the serum of 30 normal individuals and patients with either chronic (n = 30) or acute (n = 30) periapical lesions with the indirect fluorescent antibody (IFA) and enzyme-linked immunosorbent assay (ELISA) for IgG and IgM against 11 oral microorganisms routinely isolated from such lesions. The results were analyzed statistically. Analysis of variance of …

Selenium-Induced Enhancement Of Hematoporphyrin Derivative Phototoxicity In Murine Bladder Tumor Cells, Jeffrey M. Tosk

Loma Linda University Electronic Theses, Dissertations & Projects

The phototoxicity of hematoporphyrin derivative (Hpd) to murine bladder tumor (MBT-2) cells was studied in vitro. It was observed that selenium in the form of sodium selenite enhanced Hpd-sensitized photodamage in MBT-2 cells under conditions where the selenite alone was non-toxic. Sodium selenite was found to enhance the fluorescence emission of Hpd and augment the Hpd-sensitized photooxidation of tryptophan. The data indicate that sodium selenite is able to disaggregate Hpd, thereby enhancing Hpd-sensitized phototoxicity.

The Effects Of Certain Oral Bacteria On The In Vitro Growth Of Bacteroides Gingivalis, Steven G. Morrow

Loma Linda University Electronic Theses, Dissertations & Projects

The effect of nine gram positive and grain negative cocci and bacilli, which are indigenous to the oral cavity, on the in vitro growth of Bacteroides gingivalis was examined in this study. A pure culture of B. gingivalis is was obtained from the American Type Culture Collection. The nine oral test organisms were Streptococcus mitis, Streptococcus mutans, Streptococcus faecalis, Staphylococcus epidermidis, Staphylococcus aureus, Corynebacteriurn hofmanii, Lactobacillus acidophilus, Branhamella catarrhalis, and Escherichia coli. The effect on the growth of B. gingivalis of the nine test organisms was evaluated on a solid medium by use of …

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 …

The Expression Of Dna Restriction-Modification Genes Of Escherichia Coli K12, Ainu Prakash

Loma Linda University Electronic Theses, Dissertations & Projects

To facilitate the study of the expression of hsd genes in Escherichia coli K, hsdK::MudX (amp crop lac) operon fusion mutants have successfully been constructed and characterized for the first time. The results indicate that Promoters 1 and 2 for hsdK genes appear to be relatively weak, as reflected by about 50 units of beta-gal activity. Both promoters were active in all the conditions tested except during Beta-gal activity was higher in aerobic than in anaerobic growth; at 30°C and 37°C than at 42°C; and in L-broth early log phase of the culture. than in minimal media. Potential future uses …

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 …

Restriction Analysis Of Bacteriophage, Ali Reza Mostaghimi

Loma Linda University Electronic Theses, Dissertations & Projects

A restriction map of DNA from bacteriophage P3-C₁ for the endonucleases BamH I, Bgl II, EcoR I, Hind III, Sac I and Sca I is described in this study. The phage DNA was determined to be a linear double stranded molecule with a unit genomic length of 31 kb. After electrophoresis in 0.5% agarose gels, the undigested DNA appeared as three molecules of 31, 62 and 93 kb in length. The Hind III digestion pattern of the individual 31 kb molecule was identical to the 62 kb and 93 kb Hind III pattern. With all enzymes except Hind III, …

Augmentation Of Natural Killer Cell Activity By Corynebacterium Parvum: Involvement Of Lipoxygenase Pathway, Gregory D. Kuo

Loma Linda University Electronic Theses, Dissertations & Projects

The ⁵¹Chromium release assay was used to measure natural killer (NK) cell activity associated with Corynebacterium parvum (CP) injection against YAC-1 and MBT-2 targets. The peritoneal exudate cells from intraperitoneal treatment of CP at different dosages consistently gave higher cytotoxicity than their respective spleen cell fractions. In contrast, CP given intravenously showed the reversed effect. Our results demonstrate that the route of administration and dosage of CP are two crucial variables for determining the effectiveness of this immunostimulant. In vitro manipulations of nordihydroguaiaretic acid, indomethacin, and prostaglandin E2 on CP-stimulated NK cells suggest that the lipoxygenase pathway is involved …

Isolation And Characterization Of A Novel Motility Mutant In Salmonella Typhimurium, David Matthew Chacko

Loma Linda University Electronic Theses, Dissertations & Projects

A new mutant, BT40, isolated from mutagenized cultures of S. typhimurium was deficient in chemotaxis (Che^- phenotype) in tryptone semisoft agar, but had random motility with a tumbling frequency (37%) similar to that of the parental strain, ST23 (39%). In spatial assays accumulation of BT40 cells in capillary tubes containing attractants was decreased by 33%-59% compared to ST23, but BT40 had a normal response to attractants in temporal assays. The methylation level of the chemotactic proteins was decreased in BT40. At room temperature, the mean swimming speed was 11 μm/s for BT40 and 20 μm/s for ST23; more BT40 …

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 …

Assay For Rifampicin Resistant M13 Rf Synthesis, Carol Bauer Wheeler

Loma Linda University Electronic Theses, Dissertations & Projects

We have developed a rapid assay which can detect the function of the origin of 0X174 complementary strand DNA in Ml3 by virtue of its ability to initiate the synthesis of M13 complementary strands in the presence of the antibiotic rifampicin. A segment of DNA carrying the sequence recognized by the replication protein n' has been isolated from bacteriophage ØX174 and cloned into an Ml3 bacteriophage vector. Protein n' is a component of the bacterial primosome, a multi-protein assembly which initiates the synthesis of primers during DNA replication in the bacterium Escherichia coli. The recognition site for n' on the …

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 …

A Study Of Transferable Antibiotic Resistance In Enteric Bacteria, Charles H. Regeski

Loma Linda University Electronic Theses, Dissertations & Projects

A group of 48 enteric bacteria was selected from a total of 170 at Loma Linda University Medical Center because of resistance to three or more antibiotics. They were examined to see if any of the antibiotic resistances were transferred by conjugation to either of two recipient bacteria. These were a restrictionless, antibiotic sensitive, azide resistant mutant of Escherichia coli K 12 designated E. coli 1228 and a similar phenotypic strain of Salmonella typhimurium LT2 designated LB 5000. Forty-three transferred one or more antibiotic resistances to either or both recipients, a frequency of 25.3% of the total bacteria and 89.6% …

A Restriction Analysis Of Klebsiella Pneumoniae Phage Sbs, Heidi A. Walters-Khan

Loma Linda University Electronic Theses, Dissertations & Projects

A Klebsiella phage was isolated from the San Bernardino sewage and its physiochemical properties analyzed. The present study found the density of the phage to be 1.44 g/ml, the burst size to be 10, and the phage had a latent period of 20-25 minutes. The DNA was found to be double stranded and linear. Its molecular weight was 41.66 + 0.72 kilobase pairs. The DNA was digested with a battery of endoncleases and partial restriction maps determined. The potential of the phage as a vector for Klebsiella is discussed in the text.

The Effects Of Biological Response Modification And Diet On The Immune Respose Of Tumor-Bearing Mice, David Grant Lovell

Loma Linda University Electronic Theses, Dissertations & Projects

The effect that diet and other modifiers may have upon the immune system of BALB/c mice bearing tumor were studied, mice were injected s.c. with 10⁶ herpes simplex virus Type 2- Adult male BALB/c transformed (H238) cells in the thigh causing a visible fibrosarcoma within 1-2 weeks. Mice were fed either a diet low in protein (milk) and low in fat (Diet 1), or another diet low in protein (wheat) and high in fat (Diet 8). Spleen cells were collected for assays of cytotoxic T-lynphocyte (CTL) and natural killer (NK) cell activity. At 3 weeks post-injection, a decrease in …

Comparison And Correlation Of Four Methods Of Measuring Immunity To Human Cytomegalovirus, James A. Brandt

Loma Linda University Electronic Theses, Dissertations & Projects

The complement fixation test (CF) is currently the test most often used to determine the presence of antibody to human Cytomegalovirus (CMV). Several more recent techniques have now been adapted for this purpose. A comparison of CMV antibody titers was made between the CF test, a commercially available enzyme-linked immunosorbent assay (ELISA), an indirect immunofluorescent technique (IFA), and a modified indirect hemagglutination test (IHA). Forty three serum samples were tested for antibodies to CMV by each of the above procedures. The ELISA, IFA, and IHA titers were in close agreement on all samples tested. The ELISA, IFA, and IHA titers …

Mast Cell Histamine Release Induced By Portuguese Man-Of-War (Physalia) Venom, A. Lester Flowers

Loma Linda University Electronic Theses, Dissertations & Projects

Richet and Portier studied the immunization of dogs against attenuated toxic extracts of the sea anemone and showed that, when given secondary injections of sublethal amounts of native toxic extracts, the dogs experienced severe pruritus and insensitivity followed by death. They called this increased sensitivity to the toxins, anaphylaxis. Richet and Portier, and later Jacques and Schachter, and Uvnäs also studied the direct pruritic and histamine releasing effects of cnidarian (e.g. Physalia, Actinia, Cyanea), toxic extracts. While the anaphylactic effect, discovered by Richet and Portier, has been extended by numerous studies to provide the foundation of our current knowledge of …

The Effects Of Levamisole, Corynebacterium Parvum, And Unblocked Lymphocytes On Mice Injected With Herpes Virus Type 2-Transformed Cells, Linda Martin Kelln

Loma Linda University Electronic Theses, Dissertations & Projects

The association between herpes simplex virus type-2 and carcinoma of the cervix is becoming increasingly strong (Rawls, et al., 1976 ; Rapp and Duff, 1974; Hollingshead, et al-, 1973). This study demonstrates the ability of certain known immunostimulators to beneficially· alter the immune status of mice injected with HSV-2 transformed cells.

Balb/c mice were randomly divided into 6 groups. Group 1 mice were used as negative controls and received no tumor cell injection. All other groups received subcutaneous injections of 1 x 10⁶ cells/ mouse. Tumor therapy by means of irrmunostimulation was started for each group of mice 1 …

The Effects Of Temperature And Ph On The Differentiation Of Trypanosoma Lewisi (Kent, 1880) Laveran & Mesnil, 1901, In A Defined Medium, Carolann R. Rosario

Loma Linda University Electronic Theses, Dissertations & Projects

Trypanosoma lewisi is an arthropod-transmitted protozoan parasite found in the bloodstream of rats. In its life cycle it undergoes cellular differentiation. It is primarily found as the trypomastigote form in the rat and as the epimastigote form in the arthropod, the rat flea. Various studies have been done on hemoflagellates to determine the factors responsible for stimulating the differentiation from one form to the other. Included in these factors are pH, temperature, and protein fractions from fetal calf serum. Recent efforts have also been aimed at developing simple or defined media for physiological or biochemical studies on hemoflagellates. The purpose …

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 Time And Diet On The Immune Status Of Mice Challenged With Herpes-Transformed Cells, Daila S. Gridley

Loma Linda University Electronic Theses, Dissertations & Projects

Human Herpes simplex virus type 2 (HSV 2)-transformed Balb/c mouse cells (H238 cells) are malignant in Balb/c mice. The immune stratus of mice bearing H238 cell-induced tumors was studied and changes in immune response with time and with altered diet of the mice were measured.

In the experiments dealing with the status of immunity with time after tumor cell injection, mice were fed Purina Laboratory Chow (P.L.C.), injected with H238 cells, and sacrificed three to eight weeks post-injection. In the diet experiments, groups of mice were fed either P.L.C. or one of six equicaloric diets, consisting of low or high …

Non-Parental-Type Recombinants In Crosses Between Different Restriction-Modification Systems In Salmonella, Mary Ball

Loma Linda University Electronic Theses, Dissertations & Projects

Many new restriction-modification (R-M) system which are closely linked to serB and allelic to the hsd_SB system of S. typimurium occur in different Salmonella serotypes. In PI transduction studies with these systems, recombinants were isolated which fell into one of three classes with reference to their specific restriction phenotype. Recombinants that restricted like the donor, recombinants that restricted like the recipient and recombinants that had lost the ability to restrict. This latter group having lost the recipient SB restriction without gaining the specific donor restriction and here called "zero recombinants,” were of special interest and were investigated.

Zero recombinants …