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1.
Eur J Paediatr Dent ; 11(2): 82-6, 2010 Jun.
Article in English | MEDLINE | ID: mdl-20635842

ABSTRACT

AIM: To analyze the interrelationship between incisor width, deviations in the dentition and available space in the dental arch in palatally and labially located maxillary ectopic canine cases. MATERIALS AND METHODS: Size: On dental casts from 69 patients (mean age 13 years 6 months) the mesiodistal widths of each premolar, canine and incisor were measured and compared with normal standards. Dental deviations: Based on panoramic radiographs from the same patients the dentitions were grouped accordingly: Group I: normal morphology; Group IIa: deviations in the dentition within the maxillary incisors only; Group IIb: deviations in the dentition in general. Descriptive statistics for the tooth sizes and dental deviations were presented by the mean and 95% confidence limits for the mean and the p-value for the T-statistic. Space: Space was expresses by subtracting the total tooth sizes of incisors, canines and premolars from the length of the arch segments. RESULTS: Size of lateral maxillary incisor: The widths of the lateral incisors were significantly different in groups I, IIa and IIb (p=0.016) and in cases with labially located ectopic canines on average 0.65 (95% CI:0.25-1.05, p=0.0019) broader than lateral incisors in cases with palatally located ectopic canines. Space: Least available space was observed in cases with labially located canines. The linear model did show a difference between palatally and labially located ectopic canines (p=0.03). Space related to deviations in the dentition: When space in the dental arch was related to dental deviations (groups I, IIa and IIb), the cases in group IIb with palatally located canines had significantly more space compared with I and IIa. CONCLUSION: Two subgroups of palatally located ectopic maxillary canine cases based on registration of space, incisor width and deviations in the morphology of the dentition were identified.


Subject(s)
Cuspid/pathology , Dental Arch/pathology , Incisor/anatomy & histology , Malocclusion/pathology , Tooth Eruption, Ectopic/complications , Adolescent , Child , Female , Humans , Linear Models , Male , Malocclusion/complications , Maxilla , Odontometry , Tooth Abnormalities/complications , Tooth Abnormalities/pathology , Tooth Eruption, Ectopic/etiology , Tooth Eruption, Ectopic/pathology
2.
Orthod Craniofac Res ; 13(1): 34-9, 2010 Feb.
Article in English | MEDLINE | ID: mdl-20078793

ABSTRACT

OBJECTIVES: To analyse the craniofacial maxillary complex in cases with labially and palatally located ectopic canines, subgrouped accordingly: Group I: no deviations in the dentition; Group IIa: deviations in the maxillary incisors only; Group IIb: deviations in the dentition in general. SETTING AND SAMPLE POPULATION: Sixty nine patients (mean age 13 years 6 months) with palatally or labially located ectopic canines. MATERIAL AND METHODS: Profile radiographs and dental casts were analysed. The patients were subgrouped according to a previous registration of dental deviations registered radiographically. Maxillary cross-arch transversal width was analysed on dental casts. Sagittal and vertical dimensions were registered cephalometrically on profile radiographs. RESULTS: In the patient sample the maxillary cross-arch transversal width (from first maxillary molar left to first maxillary molar right), was significantly larger than the normal mean (0.65 mm, 95% Cl: 0.02-1.28, p = 0.043). The sagittal length N-S was significantly shorter (-0.97, 95% Cl:-1.72-(-)0.22, p = 0.002). The vertical length ANS-N length was also significantly shorter (-0.79, 95% Cl:-1.65-(-)0.02, p = 0.047). The remaining variables were non-significant. Tests for interaction between groups (I, IIa and IIb) and palatal/labial ectopic location did not demonstrate significance. CONCLUSION: In patients with ectopic maxillary canines, the maxillary complex is shorter sagittally as well as vertically, while it is wider transversally.


Subject(s)
Cuspid/abnormalities , Maxilla/pathology , Maxillofacial Development , Tooth Eruption, Ectopic/pathology , Cephalometry , Humans , Linear Models , Odontometry , Prognathism , Vertical Dimension
3.
Animal ; 4(10): 1709-15, 2010 Oct.
Article in English | MEDLINE | ID: mdl-22445125

ABSTRACT

This study was conducted to evaluate the effect of early rearing conditions on physiological, haematological and immunological responses relevant to adaptation and long-term stress in white Leghorn hens with intact beaks housed in furnished cages (FC) or conventional cages (CC) during the laying period. Pullets were cage reared (CR) or litter floor reared (FR). From 16 to 76 weeks of age, hens were housed in FC (eight hens per cage) or in CC (three hens per cage). As measures of long-term stress at the end of the laying period, adrenal reactivity was quantified by assessing corticosterone responses to adrenocorticotropin challenge, and immune response was assessed by measuring antibody responses after immunization with sheep red blood cells (SRBC) and keyhole limpet haemocyanin (KLH). Heterophil to lymphocyte (H/L) ratio was employed as an indicator of stress. Rearing conditions significantly affected anti-SRBC titres (P < 0.0001) and tended to affect H/L ratios (P = 0.07), with the highest values found in FR hens. Layer housing affected H/L ratio (P < 0.01); the highest ratio was found in FR birds housed in FC during the laying period. This study shows that early rearing environment affects immunological indicators that are widely used to assess stress in laying hens. However, while results on H/L ratio indicated that FR birds experienced more stress particularly when they were housed in FC during the laying period, the immune responses to SRBC in FR hens was improved, indicating the opposite. This contradiction suggests that the effects on immune response may have been associated with pathogenic load due to environmental complexity in FR and FC hens rather than stress due to rearing system or housing system per se.

4.
Clin Dev Immunol ; 10(1): 19-26, 2003 Mar.
Article in English | MEDLINE | ID: mdl-14575154

ABSTRACT

The administration of a single bolus of anti-IgM antibody to foetal lambs early in pregnancy produces prolonged B-cell depletion. The present study investigated this depletion by examining the effect, on B-cell development in the ileal Peyer's patches, of varying the timing and dosage of antibody administration and by supplementing anti-IgM with surgical splenectomy. The capacity of a 1 mg bolus of anti-IgM to deplete Peyer's patches of B cells was lost if its administration was deferred until two thirds of the way through pregnancy, but persisted beyond this time if weight-adjusted doses were used. Splenectomy of the foetus performed at an earlier age failed to extend the age at which a 1 mg dose of antibody remained effective. As the concentration of murine immunoglobulin in foetal serum was greatly reduced after 21 days, it is inferred that ongoing suppression of B-cell development is not dependent on the continued presence of murine immunoglobulin. The enduring nature of suppression could be attributable to a limited period during which differentiation of B cells from stem cells normally occurs, although further studies will be needed to investigate this and other possible explanations for the effect of anti-IgM treatment on prenatal B-cell development in sheep.


Subject(s)
Antibodies, Anti-Idiotypic/immunology , B-Lymphocytes/immunology , Fetus/immunology , Gestational Age , Immunoglobulin M/immunology , Sheep/immunology , Splenectomy , Animals , Antibodies, Anti-Idiotypic/administration & dosage , Antibodies, Monoclonal , Antigens, Differentiation, B-Lymphocyte/metabolism , B-Lymphocytes/metabolism , Dose-Response Relationship, Immunologic , Female , Immunohistochemistry , Peyer's Patches/immunology , Pregnancy , Sheep/embryology , Splenectomy/adverse effects , Splenectomy/veterinary
5.
Vet Immunol Immunopathol ; 94(3-4): 163-75, 2003 Aug 15.
Article in English | MEDLINE | ID: mdl-12909412

ABSTRACT

Anaplasma phagocytophilum infection in sheep is characterized by an immune suppression as indicated by impaired antibody response, reduced lymphocyte response and reduced oxidative burst. The effect of A. phagocytophilum infection on leucocyte populations, especially lymphocytes, was therefore investigated in six sheep experimentally infected with A. phagocytophilum, and compared with leucocyte populations from control animals.To investigate the ability of the infection to interfere with the cellular and humoral responses to specific antigens, the animals were vaccinated with commercial vaccines at the time of experimental infection, and monitored for 56 days. There were reduced percentages of gammadelta T-cells and CD4+ T-cells in peripheral blood of infected animals throughout the study period, and these cell populations showed a down-regulation of CD25 expression; while there was a relative increase in CD8+ T-cells. The reduction in CD25+ gammadelta T-cells involved a subpopulation of WC1+ gammadelta T-cells. During the first 2 weeks of the study there were reduced percentages of B-cells and leukocytes expressing MHC II and CD11b, though this decrease changed to a relative increase later in the study. The relative reductions in leucocyte populations corresponded with the observed leucopenia during the first 3 weeks post-infection, which involved lymphocyte, neutrophil and eosinophil subsets [Vet. Immunol. Immunopathol. 86 (2002) 183]. There was a reduced expression of CD11b and CD14 on granulocytes during the first 2 weeks of the study, which corresponded with the previously reported leucopenia involving neutrophils and eosinophils. Antibody responses to vaccines, lymphocyte in vitro proliferative responses to antigens and mitogens, and in vitro IFN-gamma responses to antigens were reduced up to 4 weeks after infection.


Subject(s)
Anaplasma phagocytophilum/immunology , Anaplasmosis/immunology , CD4-Positive T-Lymphocytes/immunology , CD8-Positive T-Lymphocytes/immunology , Sheep Diseases/immunology , Sheep Diseases/microbiology , Tick-Borne Diseases/veterinary , Anaplasmosis/microbiology , Animals , Antibodies, Bacterial/blood , CD4-Positive T-Lymphocytes/cytology , CD4-Positive T-Lymphocytes/microbiology , CD8-Positive T-Lymphocytes/cytology , CD8-Positive T-Lymphocytes/microbiology , Cell Division/immunology , Diphtheria Toxoid/immunology , Interferon-gamma/blood , Leukocyte Count/veterinary , Male , Random Allocation , Sheep , Tetanus Toxoid/immunology , Tick-Borne Diseases/immunology , Tick-Borne Diseases/microbiology
6.
Res Vet Sci ; 74(1): 37-45, 2003 Feb.
Article in English | MEDLINE | ID: mdl-12507565

ABSTRACT

Twenty-eight atopic dogs, 22 pruritic, non-atopic dogs and 10 healthy dogs were ELISA tested. For calculations of diagnostic specificity and sensitivity, positive ELISA test results in non-atopic dogs were considered false positive results. The absence of any positive results in the atopic dogs was considered false negative results. The atopic dogs were tested both with ELISA and an intradermal test, utilising allergen extracts from the same manufacturer, to determine the frequency of positive allergen reactions in the ELISA test compared with the intradermal test. The Prausnitz-Küstner test was performed to evaluate the significance of a positive ELISA test result. Based on cross-tabulations with clinically defined atopic dermatitis, the ELISA test showed a sensitivity of 53.6% and a specificity of 84.4%. The correlation between the ELISA and the intradermal test was poor. Positive Prausnitz-Küstner tests were not obtained using sera from dogs that were intradermal test negative for the tested allergens, even though sera had high levels of IgE as measured by the ELISA. These findings question the significance of a positive ELISA test result and indicate that the test is not measuring functional allergen-specific IgE.


Subject(s)
Allergens/immunology , Antibody Specificity , Enzyme-Linked Immunosorbent Assay/veterinary , Immunoglobulin E/analysis , Immunoglobulin E/immunology , Reagent Kits, Diagnostic/veterinary , Animals , Dermatitis, Atopic/diagnosis , Dermatitis, Atopic/immunology , Dermatitis, Atopic/veterinary , Dog Diseases/diagnosis , Dog Diseases/immunology , Dogs , Female , Male , Sensitivity and Specificity
7.
Forensic Sci Int ; 129(3): 209-13, 2002 Oct 09.
Article in English | MEDLINE | ID: mdl-12372693

ABSTRACT

We describe a method to obtain reliable mitochondrial DNA (mtDNA) sequences downstream of the homopolymeric stretches with length heteroplasmy in the sequencing direction. The method is based on the use of junction primers that bind to a part of the homopolymeric stretch and the first 2-4 bases downstream of the homopolymeric region. This junction primer method gave clear and unambiguous results using samples from 21 individuals with length heteroplasmy in the hypervariable regions HV1, HV2 or both. The method is of special value for forensic casework, because sequencing of both strands of an mtDNA region is preferable in order to reduce ambiguities in sequence determination.


Subject(s)
DNA, Mitochondrial/genetics , Genetic Variation , Sequence Analysis, DNA/methods , Haplotypes , Humans , Polymerase Chain Reaction
8.
Res Vet Sci ; 73(1): 77-85, 2002 Aug.
Article in English | MEDLINE | ID: mdl-12208110

ABSTRACT

Vaccination of goat kids against paratuberculosis protects against lesions and clinical disease. The systemic cellular response was studied in goat kids 3-9 weeks after vaccination. Peripheral blood cells showed increased interferon-gamma production and expression of interleukin-2 receptor (CD25) after stimulation with Mycobacterium avium subsp. paratuberculosis antigens. The lymph node draining the vaccination granuloma was studied three weeks after vaccination in a parallel group of goat kids. In deep cortex, MHCII+ cells were observed surrounded by CD4+ T-cells, while follicular hypertrophy and hyperplasia were prominent in the subcapsular region and along connective tissue trabecula. Comparison of the local and systemic immune responses revealed an inverse relationship between CD25+ T-cells in the lymph node deep cortex and cells in peripheral blood that up-regulate CD25 upon in vitro stimulation, suggesting that activated and regulatory T-cells in the local lymph node influence the level of circulating antigen-specific T-cells following vaccination against paratuberculosis in goats.


Subject(s)
Bacterial Vaccines/immunology , Goats/immunology , Granuloma/immunology , Histocompatibility Antigens Class II/immunology , Lymph Nodes/immunology , Mycobacterium avium subsp. paratuberculosis/immunology , Receptors, Interleukin-2/immunology , T-Lymphocytes/immunology , Animals , Antigens, Bacterial/immunology , Enzyme-Linked Immunosorbent Assay , Gene Expression Regulation , Goat Diseases/immunology , Goat Diseases/microbiology , Goat Diseases/pathology , Goat Diseases/prevention & control , Goats/microbiology , Granuloma/microbiology , Granuloma/pathology , Interferon-gamma/immunology , Interferon-gamma/metabolism , Lymph Nodes/cytology , Lymph Nodes/microbiology , Male , Mycobacterium avium subsp. paratuberculosis/isolation & purification , Paratuberculosis/immunology , Paratuberculosis/microbiology , Paratuberculosis/pathology , Paratuberculosis/prevention & control , Receptors, Interleukin-2/analysis
9.
Vet Immunol Immunopathol ; 86(3-4): 183-93, 2002 Jul.
Article in English | MEDLINE | ID: mdl-12007884

ABSTRACT

Ehrlichia phagocytophila infection in sheep is characterized by persistent neutropaenia, indicative of decreased phagocytic capacity. This predisposes infected animals to other infections. A whole blood flow cytometrical method was used to document the degree and extent of reduced phagocytic and respiratory burst activity in phagocytes during an experimental infection with E. phagocytophila, and monitored until 56 days post-infection. Six sheep at 5 months of age were inoculated with an intravenous injection of infected blood. Six age-matched sheep were used as controls. A period of reduced respiratory burst lasting up to Day 17 post-infection was recorded. The population of cells showing phagocytic activity without respiratory burst was larger in the infected animals compared to controls up to Day 45 post-infection.


Subject(s)
Ehrlichia/immunology , Ehrlichiosis/veterinary , Neutrophils/immunology , Sheep Diseases/immunology , Animals , Blood Cell Count/veterinary , Body Temperature/immunology , Ehrlichia/growth & development , Ehrlichiosis/blood , Ehrlichiosis/immunology , Ehrlichiosis/microbiology , Flow Cytometry/veterinary , Male , Neutrophils/microbiology , Phagocytosis/immunology , Random Allocation , Respiratory Burst/immunology , Sheep , Sheep Diseases/blood , Sheep Diseases/microbiology
10.
J Dairy Sci ; 85(12): 3470-5, 2002 Dec.
Article in English | MEDLINE | ID: mdl-12512620

ABSTRACT

Two groups of Norwegian Cattle showing significant genetic differences in clinical mastitis susceptibility were examined for hematological changes at three stages of lactation. Blood samples were taken from 91 healthy Norwegian Cattle cows and heifers belonging either to a high protein yield group or to a low clinical mastitis group and analyzed for hematological properties and serum cortisol levels at three stages of lactation. All animals were free of intramammary infections at the time of sampling. All cows had increased total white blood cells, neutrophils, and neutrophil/lymphocyte ratio as parturition approached, with peak levels at parturition. Cows selected for low clinical mastitis had lower levels of total white blood cells and neutrophils compared with cows selected for high protein yield throughout all three periods. The difference was significant prepartum. Cows from the low clinical mastitis group also had lower neutrophil/lymphocyte ratios and lower serum cortisol levels than those in the high protein yield group. A significant positive correlation was found between cortisol and total white blood cells and neutrophils, respectively. We conclude that selection for lower mastitis incidence in the low clinical mastitis group is associated with a significant decrease in total white blood cells and neutrophils in blood prepartum. Results from the present study, and the genetic difference in mastitis incidence observed in the groups, indicates that increased leukocyte mobilization at certain stages of lactation may be associated with increased susceptibility to mastitis.


Subject(s)
Cattle/blood , Cattle/genetics , Genetic Predisposition to Disease , Leukocyte Count , Mastitis, Bovine/genetics , Neutrophils , Animals , Female , Hydrocortisone/blood , Labor, Obstetric , Lactation , Mastitis, Bovine/blood , Norway , Pregnancy , Selection, Genetic
11.
J Microbiol Methods ; 47(3): 369-71, 2001 Dec.
Article in English | MEDLINE | ID: mdl-11714527

ABSTRACT

A Gram staining technique was developed using monodisperse magnetic beads in concentrating bacteria in suspension for downstream application. The technique does not require heat fixation of organisms, electrical power, or a microscope. Gram-negative and Gram-positive bacteria were identified macroscopically based on the colour of the suspension. The bacteria concentrated on magnetic beads may also be identified microscopically.


Subject(s)
Bacteriological Techniques , Staining and Labeling/methods , Color , Coloring Agents , Gram-Negative Bacteria , Gram-Positive Bacteria , Magnetics
12.
J Clin Microbiol ; 39(9): 3228-33, 2001 Sep.
Article in English | MEDLINE | ID: mdl-11526155

ABSTRACT

A rapid method for demonstration of gram-positive and gram-negative bacteria in milk is described. The technique is based on dilution of the sample in a medium, followed by filtration through a porous polysulfone membrane with a pore size retaining and concentrating bacteria from the sample. The bacteria concentrated on the surface of the membrane are stained with a cationic dye (toluidine blue) that can be visualized by the naked eye. After staining, the membrane is treated with ethanol-acetic acid (pH 2.8 to 3.0), which causes decolorization of gram-negative bacteria, whereas gram-positive bacteria retain the stain. The method does not require heat fixation, electrical power, microscopic examination, or specially trained personnel. The time needed to perform the test is approximately 5 min. The technique was applied to artificially infected milk and milk from cows with moderate or severe clinical mastitis for detection and differentiation of bacteria. The sensitivity of the filtration method was 92 and 100% for gram-positive and gram-negative bacteria, respectively, compared with traditional bacteriological culture of milk samples. The detection limit was 5 x 10(6) CFU/ml for Staphylococcus aureus and 1 x 10(6) CFU/ml for Escherichia coli in spiked milk samples. The overall specificity of the method was 86%. This diagnostic method can provide on-site guidance to the veterinarian to optimize use of antibiotics in mastitis therapy.


Subject(s)
Bacteriological Techniques , Gram-Negative Bacteria/isolation & purification , Gram-Positive Bacteria/isolation & purification , Mastitis, Bovine/diagnosis , Milk/microbiology , Animals , Bacteriological Techniques/instrumentation , Bacteriological Techniques/methods , Cattle , Female , Filtration/instrumentation , Filtration/methods , Gram-Negative Bacteria/classification , Gram-Negative Bacterial Infections/diagnosis , Gram-Negative Bacterial Infections/microbiology , Gram-Negative Bacterial Infections/veterinary , Gram-Positive Bacteria/classification , Gram-Positive Bacterial Infections/diagnosis , Gram-Positive Bacterial Infections/microbiology , Gram-Positive Bacterial Infections/veterinary , Mastitis, Bovine/microbiology , Membranes, Artificial , Polymers , Sulfones , Time Factors
13.
Vet Immunol Immunopathol ; 80(3-4): 271-87, 2001 Aug 10.
Article in English | MEDLINE | ID: mdl-11457480

ABSTRACT

An experimental oral infection of goats with a caprine isolate of Mycobacterium a. subsp. paratuberculosis was used to investigate immunological and bacteriological events during the subclinical phase of infection. Seven goats at 5-8 weeks of age were given a bacterial suspension in milk-replacement three times weekly for 9 weeks. Six animals were kept as controls. Cellular recall responses against M. a. paratuberculosis were analysed by means of a lymphocyte proliferation test, an IFN-gamma assay and an IL-2 receptor assay. All inoculated animals had detectable CMI responses from 9 weeks post-inoculation and through the 2 years of study, although the responses were highest during the first year. Antibodies against M. a. paratuberculosis could be detected from weeks 15-20 in four of the seven animals, and one additional animal became antibody positive at week 35, while two inoculated animals did not produce significant antibody titres during the experiment. At about 1-year post-inoculation, two animals became faecal shedders, while two others started to excrete bacteria into faeces about 2 years post-inoculation. The appearance of M. a. paratuberculosis in faeces was not associated with a decline in cellular responses as far as could be assessed using the current methods for measuring CMI. Pathological lesions due to M. a. paratuberculosis infection and presence of bacteria were recorded in the intestine and/or mesenteric lymph nodes of five animals while lymph node changes suggestive of paratuberculosis were observed in one animal. Only the two animals with no signs of an active infection at necropsy showed a considerable decline in the cellular parameters during the last year of the study, particularly in the IFN-gamma assay. The two animals with the highest levels of M. a. paratuberculosis responsive CD8+ lymphocytes in the circulation about 1-year post-inoculation had no detectable lesions in the distal ileum and colon at necropsy, while high numbers of gammadelta T-cells responsive to M. a. paratuberculosis in the circulation were associated with disseminated lesions in the distal ileum and colon.


Subject(s)
Goat Diseases/immunology , Goat Diseases/microbiology , Paratuberculosis/immunology , Animals , Antibodies, Bacterial/biosynthesis , Feces/microbiology , Goat Diseases/pathology , Goats , Immunity, Cellular , In Vitro Techniques , Interferon-gamma/biosynthesis , Lymphocyte Activation , Lymphocyte Subsets/immunology , Male , Mycobacterium avium subsp. paratuberculosis/immunology , Mycobacterium avium subsp. paratuberculosis/isolation & purification , Paratuberculosis/microbiology , Paratuberculosis/pathology , Receptors, Interleukin-2/metabolism
14.
J Mol Biol ; 307(1): 67-74, 2001 Mar 16.
Article in English | MEDLINE | ID: mdl-11243804

ABSTRACT

Upon binding of a decamer bis-PNA (H-Lys-TTCCTCTCTT-(eg1)(3)-TTCTCTCCTT-LysNH(2)) to a complementary target in a double-stranded DNA fragment, three distinct complexes were detected by gel mobility shift analysis. Using in situ chemical probing techniques (KMnO(4) and DMS) it was found that all three complexes represent bona fide sequence-specific PNA binding to the designated target, but the complexes were structurally different. One complex that preferentially formed at higher PNA concentrations contains two bis-PNA molecules per DNA target, whereas the other two complexes are genuine triplex invasion clamped structures. However, these two latter complexes differ by the path relative to the DNA target of the flexible ethylene-glycol linker connecting the two PNA oligomers that comprise a bis-PNA. We distinguish between one in which the linker wraps around the non-target DNA strand, thus making this strand part of the triplex invasion complex and another complex that encompass the target strand only. The implications of these results are discussed in terms of DNA targeting by synthetic ligands.


Subject(s)
DNA/chemistry , Peptide Nucleic Acids/chemistry , DNA/chemical synthesis , DNA/metabolism , Electrophoresis , Isomerism , Nucleic Acid Conformation , Peptide Nucleic Acids/chemical synthesis , Peptide Nucleic Acids/metabolism , Plasmids/genetics
15.
Vet Immunol Immunopathol ; 77(1-2): 43-54, 2000 Nov 23.
Article in English | MEDLINE | ID: mdl-11068065

ABSTRACT

Quantification of surface IL-2R expression on activated lymphocytes by flow cytometry have recently been reported to be useful in measuring cellular immunity against Mycobacterium avium subsp. paratuberculosis in goats (Whist et al., 2000, Vet. Immunol. Immunopathol. 73, 207-218). To characterise the phenotype of the peripheral lymphocytes expressing IL-2R after in vitro stimulation with purified protein derivative (PPD) from M. a. paratuberculosis, cells were processed for dual or triple colour analysis by flow cytometry (CD4 and IL-2R or CD8, gammadelta-TcR and IL-2R). To distinguish the response of antigen-specific T cells from non-specific stimulation, we performed a time-course study of proliferating cells in a group of M. a. paratuberculosis-infected animals and a control group. Following in vitro stimulation with PPD of whole blood for three different periods of time, IL-2R expression was detected mainly not only in gammadelta-T cells, but also in CD4+ and CD8+ T cells. We found a specific response of gammadelta-T cells from infected animals after 24h of stimulation. Following 120h of stimulation, however, gammadelta-T cells from control animals up-regulated IL-2R to the same level as those from infected animals, indicating either a non-specific stimulation or activation due to a first line of defence against Mycobacterium antigens. The CD4+ cells showed a specific response to PPD stimulation at all three time points. A minor population of antigen reactive gammadelta+ cells also expressed CD8. The proliferative responses differed between alphabeta and gammadelta-T cells; the IL-2R+ alphabeta T cell population mainly comprised proliferating cells, while the gammadelta+ population showed less expansion.


Subject(s)
Goat Diseases/immunology , Paratuberculosis/immunology , Receptors, Interleukin-2/metabolism , T-Lymphocyte Subsets/immunology , Animals , CD4-Positive T-Lymphocytes/immunology , CD8-Positive T-Lymphocytes/immunology , Goats , In Vitro Techniques , Kinetics , Lymphocyte Activation , Male , Receptors, Antigen, T-Cell, gamma-delta/metabolism , Tuberculin
16.
J Toxicol Environ Health A ; 59(7): 561-74, 2000 Apr 14.
Article in English | MEDLINE | ID: mdl-10777247

ABSTRACT

Associations between immunoglobulin G (IgG) levels and the organochlorine contaminants (OCs) polychlorinated biphenyls (PCBs), chlordanes, 1,1-dichloro-2,2-bis(4-chlorophenyl) ethylene (DDE), hexachlorobenzene (HCB), and hexachlorocyclohexanes (HCHs) in blood plasma from polar bears caught at Svalbard were determined. The blood samples were collected from free-living polar bears of different age and sex between 1991 and 1994. The IgC concentration increased with age and was significantly higher in males than in females. IgG was negatively correlated with sigmaPCB level and with three individual PCB congeners, IUPAC numbers 99, 194, and 206. HCB was also negatively correlated with IgG. The significant negative OC correlation with IgG levels may indicate an immunotoxic effect.


Subject(s)
Environmental Pollutants/toxicity , Immunity/drug effects , Insecticides/toxicity , Polychlorinated Biphenyls/toxicity , Ursidae/physiology , Aging/metabolism , Animals , Environmental Pollutants/blood , Female , Immunoglobulin G/immunology , Immunoglobulin G/metabolism , Insecticides/blood , Male , Norway , Polychlorinated Biphenyls/blood , Seasons
17.
Vet Immunol Immunopathol ; 73(3-4): 207-18, 2000 Mar 15.
Article in English | MEDLINE | ID: mdl-10713335

ABSTRACT

The purpose of the present work was to demonstrate cell-mediated immune response to paratuberculosis in experimentally infected animals, using quantification of interleukin-2 receptor (IL-2R) expression on activated lymphocytes by means of in vitro stimulation with Mycobacterium avium ssp. paratuberculosis-derived purified protein derivative (PPDp). A whole-blood technique was developed, and optimal conditions for quantification of IL-2R expression on caprine lymphocytes, using monoclonal antibodies (anti-bovine IL-2R-alpha) and low cytometrical analysis, were determined. Different PPDp-antigen concentrations and incubation times were compared. The whole-blood method was also compared to the more traditional IL-2R assay using peripheral blood mononuclear cultures (Hesketh et al., 1993). Cross-reactivity to Mycobacterium avium was studied at different mycobacteria-PPD concentrations. An immune response could be demonstrated in animals infected with Mycobacterium avium ssp. paratuberculosis. We found that a PPDp concentration of 10microgml(-1) together with an incubation time of 72h, gave the best results using the whole-blood method. The whole-blood method eliminates many laborious steps involved in lymphocyte separation, and the effects of all the constituents of blood are expressed in a way which corresponds more to in vivo conditions. The risk of selecting subpopulations of lymphocytes during cell separation is avoided.


Subject(s)
Goat Diseases/immunology , Mycobacterium avium subsp. paratuberculosis/immunology , Paratuberculosis/immunology , Receptors, Interleukin-2/biosynthesis , T-Lymphocytes/immunology , Animals , Antibodies, Bacterial/analysis , Antibodies, Monoclonal , Antigens, Bacterial/pharmacology , Biomarkers/analysis , Concanavalin A/pharmacology , Flow Cytometry/veterinary , Goats , Immunity, Cellular , Lymphocyte Activation/drug effects , Male , Mycobacterium avium/immunology
18.
Toxicol Lett ; 112-113: 103-9, 2000 Mar 15.
Article in English | MEDLINE | ID: mdl-10720718

ABSTRACT

Alarmingly high polychlorinated biphenyl (PCB) levels have been found in the top predators such as glaucous gull (Larus hyperboreus) and polar bear (Ursus maritimus) at Svalbard [Gabrielsen, G.W., Skaare, J.U., Polder, A., Bakken, V., 1995. Chlorinated hydrocarbons in glaucous gull (Larus hyperboreus). Sci. Total Environ. 160/161, 337-346; Bernhoft, A., Skaare, J.U., Wiig, O., 1997. Organochlorines in polar bears (Ursus maritimus) at Svalbard. Environ. Pollut. 95, 159-175; Henriksen, E.O., Gabrielsen, G.W., Trudeau, S., Wolkers, H., Sagerup, K., Skaare, J.U., 1999. Organochlorines and possible biochemical effects in glaucous gull (Larus hyperboreus) from Bear Island, the Barents Sea. Arch. Environ. Contam. Toxicol. (in press). ]. Studies of the possible toxic effects, particularly on the immune system and reproduction, of the very high PCB levels in these species are currently being investigated. Data obtained in the field (f.i. reproductive success in polar bears and intestinal nematodes in glaucous gulls), as well as levels of various biochemical and physiological parameters (f.i. thyroid hormones, retinol, EROD activity, CYP1A, IgG), have been coupled with the PCB levels [Skaare, J.U., Wiig, O., Bernhoft, A., 1994. Klorerte organiske miljogifter; Nivâer og effekter i isbjorn. Norwegian Polar Institute Reportseries no. 86, 1-23 (in Norwegian); Bernhoft, A., Skaare, J.U., Wiig, O., 1997. Organochlorines in polar bears (Ursus maritimus) at Svalbard. Environ. Pollut. 95, 159-175; Bernhoft, A., Skaare, J.U., Wiig, O., Derocher, A.E., Larsen, H.J., 2000. Possible immunotoxic effects of organochlorines in polar bears (Ursus maritimus) at Svalbard (in press); Henriksen, E.O., Gabrielsen, G.W., Skaare, J.U., Skjegstad, N., Jensen, B.M., 1998a. Relationship between PCB levels, hepatic EROD activity and plasma retinol in glaucous gull, Larus hyperboreus. Marine Environ. Res. 46, 45-49; Henriksen, E.O., Gabrielsen, G.W., Trudeau, S., Wolkers, H., Sagerup, K., Skaare, J.U. , 1999. Organochlorines and possible biochemical effects in glaucous gull (Larus hyperboreus) from Bear Island, the Barents Sea. Arch. Environ. Contam. Toxicol. (in press); Sagerup, K., Gabrielsen, G.W., Skorping, A., Skaare, J.U., 1998. Association between PCB concentrations and intestinal nematodes in glaucou gulls, Larus hyperboreus, from Bear Island. Organohalogen compounds 39, 449-451; Skaare, J.U., Wiig, O., Bernhoft, A., 1994. Klorerte organiske miljogifter; Nivâer og effekter i isbjorn. Norwegian Polar Institute Reportseries no. 86, 1-23. (in Norwegian)].


Subject(s)
Birds/metabolism , Environmental Pollutants/toxicity , Polychlorinated Biphenyls/toxicity , Ursidae/metabolism , Adipose Tissue/chemistry , Animals , Cytochrome P-450 CYP1A1/metabolism , Environmental Monitoring , Environmental Pollutants/analysis , Female , Gas Chromatography-Mass Spectrometry , Immune System/drug effects , Immunoglobulin G/blood , Male , Polychlorinated Biphenyls/analysis , Svalbard , Vitamin A/blood
19.
Biochim Biophys Acta ; 1489(1): 159-66, 1999 Dec 10.
Article in English | MEDLINE | ID: mdl-10807005

ABSTRACT

Peptide nucleic acid (PNA) is a nucleic acid mimic in which the deoxyribose phosphate backbone has been replaced by a pseudo-peptide polymer to which the nucleobases are linked. PNA-oligomers can be synthesized in relatively large amounts, are highly stable in biological environments, and bind complementary DNA and RNA targets with remarkably high affinity and specificity. Thus PNA possesses many of the properties desired for a good antisense agent. Until recently, limited uptake of PNA into cells has been the major obstacle for applying PNA as an antisense agent in cell cultures and in vivo. Here, the antisense properties of PNA in vitro and in vivo will be reviewed. In particular, we will focus on recent observations indicating that PNA equipped with or without various uptake moieties may function as an efficient and gene-specific inhibitor of translation in Escherichia coli and in certain mammalian cell types.


Subject(s)
Oligonucleotides, Antisense/chemistry , Peptide Nucleic Acids/chemistry , Animals , Cell Membrane Permeability , Cells, Cultured , Drug Carriers/metabolism , Drug Stability , Escherichia coli/metabolism , Neurons/cytology , Neurons/drug effects , Neurons/enzymology , Nucleic Acid Hybridization/drug effects , Oligonucleotides, Antisense/pharmacology , Oxytocin/antagonists & inhibitors , Oxytocin/genetics , Peptide Nucleic Acids/pharmacology , Protein Biosynthesis/drug effects , RNA, Messenger/antagonists & inhibitors , RNA, Messenger/biosynthesis , RNA, Messenger/genetics , Rats , Receptors, Galanin , Receptors, Neuropeptide/antagonists & inhibitors , Receptors, Neuropeptide/genetics , Receptors, Neurotensin/antagonists & inhibitors , Receptors, Neurotensin/genetics , Receptors, Opioid, mu/antagonists & inhibitors , Receptors, Opioid, mu/genetics , Ribonuclease H/metabolism
20.
Genet Res ; 71(3): 187-91, 1998 Jun.
Article in English | MEDLINE | ID: mdl-9717438

ABSTRACT

There are two known alleles of the mouse parotid secretory protein (PSP) gene: Pspa and Pspb. Pspa is carried by DBA/2J mice and Pspb is carried by C57BL/6J mice. Eighty-eight mice derived from a F1(C57BL/6J x DBA/2J) to DBA/2J backcross were analysed for PSP mRNA expression in the sublingual glands. Expression was found in heterozygous mice only. This indicates that only Pspb is expressed in this tissue. Furthermore, it maps the allele-specific sublingual gland determinant within 3.4 cM of Psp. Previous analysis of Pspb identified an enhancer-like region in position -4.6 to -3.1 kb that was necessary for transgene expression in the sublingual glands. Here it is shown that the corresponding region in Pspa enhances transgene expression in the sublingual glands as efficiently. The implications for regulation of PSP mRNA expression in the sublingual glands are discussed.


Subject(s)
Gene Expression Regulation/genetics , Salivary Proteins and Peptides/genetics , Sublingual Gland/physiology , Alleles , Animals , Crosses, Genetic , Enhancer Elements, Genetic/genetics , Mice , Mice, Transgenic
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