Search PubMedSearch

Biomedical subjects

D P Anderson

Publications and source records attributed to D P Anderson.

At least 19 recordsLinked to original sources

Neutrophil, glass-adherent, nitroblue tetrazolium assay gives early indication of immunization effectiveness in rainbow trout.

Neutrophil activity in rainbow trout (Oncorhynchus mykiss) is increased upon antigenic stimulation with the Yersinia ruckeri O-antigen bacterin. The characteristics of neutrophil attachment to glass and nitroblue tetrazolium (NBT) staining were used to determine the effectiveness of immunization programs with fingerling rainbow trout. Fish immunized by intraperitoneal injection with doses of 100, 10, or 1 microgram of the bacterin showed the highest responses in that order in numbers of glass adherent, NBT-positive neutrophils. Studies on the kinetics of the occurrence of numbers of glass-adherent, NBT-positive staining cells from the fish injected with the 10 micrograms dose showed the numbers of positive cells were largest on Day 2 after injection. The specific immune response was confirmed by demonstrating the presence of plaque-forming cells by the passive hemolytic plaque assay and the rise in humoral antibody titers by passive hemagglutination 12 days after injection. The effects of immunization in trout could be detected earlier by using the neutrophil glass adherence and NBT reduction assays than by using assays based on observations of the specific immune response.

Animals

Immunostimulants added to injected Aeromonas salmonicida bacterin enhance the defense mechanisms and protection in rainbow trout (Oncorhynchus mykiss).

Immunostimulants were given to rainbow trout for assaying effects on modulating non-specific defense mechanisms, specific immune response, and protection levels against pathogen challenge with Aeromonas salmonicida. Three drugs, levamisole (an approved veterinary drug in the USA), a quaternary ammonium compound (QAC), and a short-chain polypeptide (ISK) were found to affect the non-specific defense mechanism activities, which were measured by changes in circulatory neutrophil and phagocytic activity levels, and the specific immune response factors, which were measured by numbers of plaque-forming cells, and circulatory antibody levels. When given alone, the immunostimulants elevated the non-specific factors. When injected in combination with an A. salmonicida O-antigen bacterin, the non-specific factors were further elevated, and the specific response was raised over samples taken from fish given the bacterin without the immunostimulants. Challenge tests with the virulent pathogen, A. salmonicida, showed a 5-6 day delay in the onset of mortalities in the fish given the immunostimulants alone, and a 12-14 day delay when immunostimulants given were combined with the bacterin. In the groups given the QAC or ISK with the bacterin, there was a 20% and 40% survival rate, respectively.

Adjuvants, Immunologic

In vitro effect of levamisole on the neutrophil activity in rainbow trout (Oncorhynchus mykiss).

An in vitro immunization and cultivation method for fish spleen organ section was used to investigate the effects of levamisole on the neutrophil activity. After 10 days of culture with either 50 micrograms/ml, 25 micrograms/ml, 5 micrograms/ml or no levamisole in the media, the nonspecific defense reactions were measured by determining the metabolic activity of neutrophils by using the nitroblue tetrazolium test, and phagocytic and adherence indexes by incubating the fish cells with suspensions of formalin-killed Staphylococcus aureus. Elevations were found in nonspecific cellular defense in the 5 micrograms/ml levels of levamisole. The 25 micrograms/ml levels instigated a slight elevation in nonspecific cellular defense and the 50 micrograms/ml levels suppressed all indicators, whether levamisole was given alone or combined with Yersinia ruckeri O-antigen or DNP-Ficoll. These antigens are themselves nonspecific cellular defense stimulators, and a synergistic effect was evident when combined with 25 micrograms/ml and 5 micrograms/ml levels of levamisole.

Animals

In vitro immunostimulation of rainbow trout (Oncorhynchus mykiss) spleen cells with levamisole.

An in vitro immunization and cultivation method for fish spleen organ sections was used to investigate the effects of levamisole on the immune response. After 10 days of culture with either 50 micrograms/ml, 25 micrograms/ml, 5 micrograms/ml or no levamisole in the media, the nonspecific defense reactions were measured by determining the metabolic activity of neutrophils by using the nitroblue tetrazolium test, and phagocytic and adherence indexes by incubating the fish cells with suspensions of formalin-killed Staphylococcus aureus. The specific immune system activity was shown by the passive hemolytic plaque assay demonstrating the numbers of antibody-producing cells. Elevations were found in nonspecific defense and specific immune response in the 5 micrograms/ml levels of levamisole. The 25 micrograms/ml levels instigated a slight elevation in some nonspecific levels but showed suppression in the specific immune response. The 50 micrograms/ml levels suppressed all indicators whether levamisole was given alone or combined with Yersinia ruckeri O-antigen or DNP-Ficoll. These antigens are themselves nonspecific defense stimulators, and a synergistic effect was evident when combined with 25 micrograms/ml and 5 micrograms/ml levels of levamisole.

Animals

Comparisons of nonspecific and specific immunomodulation by oxolinic acid, oxytetracycline and levamisole in salmonids.

Oxolinic acid, a promising drug for the treatment of bacterial fish disease agents, was tested for possible immunomodulatory effects on fish. Another antibiotic oxytetracycline, known to be immunosuppressive at higher treatment doses, and levamisole, a known immunostimulator for higher vertebrates, were also compared for causing changes in the nonspecific defense compartment and the specific immune system in rainbow trout. Groups of fish were immunized with Yersinia ruckeri O-antigen bacterin in combination with selected doses of the drugs. The nonspecific defense activity was measured by demonstrating neutrophil metabolic activity by the nitroblue tetrazolium assay, by counting engulfed bacterial cells for a phagocytic index and by counting leukocytes with adherent bacterial cells for the adherence index. The specific immune response was monitored by the passive hemolytic plaque assay demonstrating the numbers of antibody-producing cells. The results showed that oxolinic acid, used at recommended doses for the treatment of bacterial diseases, did not cause immunosuppression in either the nonspecific defense or specific immune system compartments, whereas tetracycline at 10 mg/kg caused reduced activity in both. Fish given levamisole injections before the antigen injection showed a stimulated nonspecific defense but a much reduced specific immune response.

Adjuvants, Immunologic

Vitamin A concentrations in liver determined by isotope dilution assay with tetradeuterated vitamin A and by biopsy in generally healthy adult humans.

The vitamin A status in 11 generally healthy surgical patients was estimated by measuring the dilution of a 45-mg oral dose of tetradeuterated retinyl acetate (99% pure). After purification of retinol by high-performance liquid chromatography, the ratio of 2H4-retinol:1H-retinol in plasma was measured by gas chromatography-mass spectrometry. On the basis of the observed ratios of [2H4]retinol:[1H]retinol over 19-47 d, the total body reserves and liver concentrations of vitamin A were calculated. Liver biopsy samples taken at surgery were directly analyzed for vitamin A. The correlation coefficient between calculated and measured liver vitamin A concentrations for 10 of the subjects was 0.88, and the Spearman's rank correlation coefficient was 0.95 (p less than 0.002). Thus, total body reserves of vitamin A in humans can be estimated validly in the marginal and satisfactory ranges by a benign, relatively noninvasive procedure.

Adult

Methylmercury and the health of autochthons in northwest Quebec.

We conducted a cross-sectional epidemiologic and clinical study to evaluate the nature, magnitude, and frequency of unfavourable health states among residents of northwest Quebec. Of particular interest were the possible occurrence of Minamata disease and any other neurological abnormalities. We also sought to determine whether industrial or naturally-occurring mercury in the region's environment were causally associated with any disorders that might be detected. A total of 321 subjects were studied in four groups with contrasting exposure to point-source mercury from a chlor-alkali plant in the area. Methylmercury in hair served as the marker of subject's ingestion of any organic mercury (natural or industrial). Ten pre-chosen target variables, seven clinical and three neurophysiological, were assessed in all subjects. Relationships between mercury in hair and each target variable were calculated. We determined whether gradients in target variables corresponded to gradients in potential exposure of the subjects to the aquatic emissions of the chlor-alkali plant. Minamata disease was not found. Diabetes and alcohol-related health problems assessed as important clinically plausible confounders, were common. Differences in frequency or magnitude of clinical, toxicological, and physiological findings among the contrast groups were small and not of clinical or statistical significance. There were definite relationships between mercury in hair and four target variables despite the low values of mercury measured and the mild nature of the health problems observed. This would suggest that verifiable biological threshold levels for clinical effects, if such exist, may be substantially lower among Canadians than those currently adopted as policy for adults elsewhere. A gradient was found only for two of the ten target variables. Clinically meaningful alternate hypotheses for observed health effects emerge in the data and agree with observations in the field made by consulting clinical experts.

Animals

Water imbibition in the rat prostate during castration-induced regression.

The present study was conducted to examine changes in water concentration in the ventral prostate of adult rats at different intervals following castration. The prostatic dry weight was obtained by drying the fresh prostate at 70 degrees C for at least 110 hr and the prostatic water content was calculated from its wet and dry weight. Under normal conditions, the prostatic water concentration ranges from 81.9 to 82.7% in uncastrated rats. The prostatic water concentration started to increase at 8 hr postcastration (83.1%) but this increase was not statistically significant. By 16 hr postcastration, the prostatic water concentration (83.8%) became significantly higher than that of the uncastrated animals. In rats of day 1 to day 10 postcastration, the prostate contained significantly more water (84.4%-84.7%) than those in uncastrated animals. By day 21 postcastration, the prostatic water concentration (81.5%) was not significantly different from that of uncastrated rats. Unlike the prostate, the skeletal muscle did not significantly change the water concentration following castration. The 51Cr-EDTA space in the prostate was not significantly different in rats before and after castration. These results indicate that water imbibition in the rat prostate is associated with an active period of prostatic regression and that the change in the 51Cr-EDTA space during prostatic regression is not the major cause of water imbibition in the tissue. Therefore, the present findings suggest that castration-induced water imbibition in the rat prostate is caused by an increase in the intracellular water space.

Adrenalectomy

Immunization and culture of rainbow trout organ sections in vitro.

Splenic and anterior kidney sections or whole organs were excised from large (1 kg) or small (200 g) rainbow trout (Salmo gairdneri) and placed in sterile 60 mm plastic plates containing 10 ml of Eagle's minimal essential medium (EMEM) supplemented with normal or fetal calf serum for in vitro culture. The organ samples were immunized in vitro by direct injection or by mixing in the medium Yersinia ruckeri O-antigen or dinitrophenyl-Ficoll. The medium was changed once during the 10-day incubation at 15 C. The passive hemolytic plaque assay demonstrated antibody production from the plaque-forming cells (PFC); passive hemagglutination was used to measure antibody titers in the media. High numbers of PFC occurred in cultures of either kidney or spleen, demonstrating that these organs can function independently for antibody production. Splenic sections from large fish produced more PFC than comparable whole organs from small fish. EMEM supplemented with 2% normal calf serum was a satisfactory culture medium. 2-hydroxyethyl-mercaptan an ingredient used in mammalian cell culture, inhibited antibody production in trout cells. These techniques are being used in the culture of organs and cells to elucidate pathways and sequences of antigen uptake and delivery of the immunopoietic tissues in trout.

Animals

Effect of exogenous corticosteroids on circulating virus and neutralizing antibodies in striped bass (Morone saxatilis) infected with infectious pancreatic necrosis virus.

Corticosteroids have been reported to induce immunosuppression in fish exposed to many types of bacterial antigens. We document a similar phenomenon in fish exposed to infectious pancreatic necrosis virus (IPNV). Fingerling striped bass that were injected with the steroid triamcinolone acetonide (100 mg/kg body weight) 24 hours before receiving intraperitoneal inoculation with IPNV became viremic 3 days post inoculation (dpi) and virus was still detected in the buffy coat cells 14 dpi. In contrast, viremia could not be detected after 7 dpi in fish that received virus but not steroids. Circulating virus neutralizing antibodies were first detected in steroid treated fish at 10 dpi compared to 7 dpi for the virus injected fish and titers were consistently lower in the steroid group. Steroid treatment of chronic IPNV-carriers did not induce detectable viremia nor alter circulating antibody levels in chronic IPNV-carriers. None of the striped bass demonstrated clinical signs of viral disease.

Animals

Visual abnormalities with multiple trauma.

The diversity of pathogenetic mechanisms involved in posttraumatic visual impairment was reviewed in a study of the hospital records of 24 patients admitted with multiple injuries. Most major visual abnormalities occurred in young people (average age 33 years) who presented with a wide range of overall severity of injury (injury severity score 13-47) and involvement of the central nervous system (Glasgow coma scale 5-15). Bilateral or monocular blindness developed in 63% of patients. Seventy percent of the injuries involved the anterior visual pathways with damage to the optic nerve alone accounting for 35%. Fractures of the sphenoid bone, particularly of the body, accompanied optic nerve and chiasmal injuries and some cases of traumatic carotid-cavernous fistulas. Pathogenetic mechanisms varied according to the site of injury and included vitreous hemorrhage and optic atrophy secondary to raised intracranial pressure, retinal hypoxia from carotid-cavernous fistulas, shearing and compression injuries of the optic nerve, traumatic chiasmal syndrome, temporoparietal and occipital contusions, and transtentorial herniation with occipital infarction. Visual abnormalities varied in severity from moderately reduced visual acuity and diverse hemianopias and scotomas to blindness. The incidence of posttraumatic residual visual abnormalities is likely to increase in the wake of improved acute care of the traumatized victim.

Adolescent

Molecular analysis of the lymphocyte membrane.

Methods for the molecular analysis of lymphocyte membranes are reviewed briefly, and wherever possible presented in a manner relevant to comparative studies. The specific areas reviewed include the bulk isolation of lymphocyte membranes, the use of radioisotopes to covalently label lymphocyte membrane molecules, the use of lectins to characterize membrane glycoconjugates, and our current understanding of lymphocyte membrane immunoglobulins.

Animals

The immune response in immunized and naturally infected rainbow trout (Salmo gairdneri) to Diplostomum spathaceum as detected by enzyme-linked immunosorbent assay (ELISA).

The enzyme-linked immunosorbent assay (ELISA) procedure was modified and adapted for detection of circulating antibodies in rainbow trout (Salmo gairdneri) against metacercariae of the digenean trematode Diplostomum spathaceum, the causative agent for diplostomiasis. Rainbow trout (Salmo gairdneri) were injected with sonicated metacercariae representing 10, 40, and 100 metacercariae per fish. Three weeks after immunization the average titers for trout injected with 10, 40, and 100 metacercariae were 874, 841, and 525, and by six weeks the titers had fallen to 299, 349, and 203, respectively. Nine weeks after initial immunization, two remaining fish initially immunized with 100 metacercariae per fish were injected with a booster of 50 sonicated metacercariae per fish. Four weeks later the average titer was 1204. Serum samples from naturally infected wild fish tested for the presence of circulating antibodies against Diplostomum spathaceum showed 25 of 27 with positive titers.

Animals

Flush exposure and injection immunization of rainbow trout to selected DNP conjugates.

Rainbow trout (Salmo gairdneri) were immunized by flush exposure or intraperitoneal injection with single doses of the following dinitrophenyl (DNP) conjugates: DNP-keyhole limpet hemocyanin (KLH), DNP-Ficoll, DNP-O-antigen (from the fish pathogen, Yersinia ruckeri), DNP-sheep red blood cells, or DNP-duck red blood cells. The immune response was demonstrated by the passive hemolytic plaque assay to show splenic antibody-producing cells (APC) 14 days after antigen administration. High numbers of splenic APC specific for the hapten were found when the conjugates DNP-KLH, DNP-Ficoll, and DNP-O-antigen were given by injection and when DNP-O-antigen was given by flush exposure. The heterologous red blood cells were relatively nonimmunogenic. When the immune response to the hapten and carrier could be measured--e.g., after immunization with the DNP-O-antigen--the numbers of APC were consistently higher for the hapten.

Animals

Thymocyte plasma membrane of the rainbow trout, Salmo gairdneri: associated immunoglobulin and heteroantigens.

Thymic lymphocytes of the rainbow trout, S. gairdneri were disrupted and a plasma membrane containing fraction isolated by differential and buoyant density centrifugation. Radioiodine introduced into the membrane by the lactoperoxidase catalyzed reaction and immunoglobulin (identified by radioimmunoassay with monoclonal antibody) both copurified in the plasma membrane fraction. Rabbit antibody raised to the plasma membrane fraction showed a strong reaction with trout lymphocytes in immunofluorescence, was mitogenic for trout lymphocytes, and recognized lymphocyte membrane heteroantigens of molecular weight greater than 70,000 in the thymus and 45,000-95,000 in the head kidney.

Animals

Abnormal blood vessels on the optic disc.

Twenty-five patients with abnormal blood vessels on the optic nerve head were examined both clinically and with fluorescein angiography to determine the diagnostic significance of these abnormal vessels. Six patients had congenital abnormal vessels--prepapillary or optociliary arteries or veins, or vessels associated with congenital anomalies of the disc. Nineteen patients had acquired abnormal vessels that were associated with retinal vein occlusion, chronic glaucoma, chronic papilledema, meningioma or vasculitis. The clinical and fluorescein angiographic features that differentiated these groups are discussed in detail.

Adult