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Biomedical subjects

J A Ellis

Publications and source records attributed to J A Ellis.

At least 73 records · Page 4Linked to original sources
PubMed1998

Clinical and immunologic responses of vaccinated and unvaccinated calves to infection with a virulent type-II isolate of bovine viral diarrhea virus.

OBJECTIVE: To determine efficacy of a modified-live type-I isolate of bovine viral diarrhea virus (BVDV) vaccine in protecting calves from infection with a virulent type-II isolate, and to determine which type of immune response (i.e., humoral or cellular) correlates with protection. DESIGN: Prospective study. ANIMALS: 28 neonatal Holstein and Holstein-cross calves. PROCEDURE: Within 18 hours of birth, calves received maternal colostrum or were fed pooled colostrum. On days 7 to 10 after birth, calves were determined to be seropositive (n = 16) or seronegative (12) for antibodies to BVDV on the basis of ELISA and virus neutralization test results. Seropositive and seronegative 10- to 14-day-old calves were then given a combined vaccine that contained a modified-live type-I isolate of BVDV or a similar vaccine that lacked protection against bovine viral diarrhea. All calves were inoculated intranasally approximately 21 days after vaccination with a virulent type-II isolate of BVDV. Clinical and immunologic variables, including clinical scores, rectal temperatures, results of CBC with lymphocyte subset analysis, antibody responses, and cell-mediated immune responses, were monitored for 14 days after inoculation. RESULTS: Seronegative-unvaccinated calves developed severe disease and required euthanasia. Vaccination of seronegative calves with a modified-live type-I isolate had a disease-sparing effect as did passive transfer of colostral antibodies to BVDV. Clinical scores were not significantly different between seropositive-vaccinated and seropositive-unvaccinated calves after viral inoculation. CLINICAL IMPLICATIONS: A single dose of a modified-live type-I isolate of BVDV vaccine protects young calves from clinical signs of disease associated with type-II isolates.

Animals↗
PubMed1998

Genetic analysis of male pattern baldness and the 5alpha-reductase genes.

Genetic predisposition and androgen dependence are important characteristics of the common patterned loss of scalp hair known as male pattern baldness. The involvement of the 5alpha-reductase enzyme in male pattern baldness has been postulated due to its role in the metabolism of testosterone to dihydrotestosterone. There are two known isozymes of 5alpha-reductase. Type I has been predominantly localized to the skin and scalp. Type II, also present on the scalp, is the target of finasteride, a promising treatment for male pattern baldness. We conducted genetic association studies of the 5alpha-reductase enzyme genes (SRD5A1 on chromosome 5 and SRD5A2 on chromosome 2) using dimorphic intragenic restriction fragment length polymorphisms. From a population survey of 828 healthy families comprising 3000 individuals, we identified 58 young bald men (aged 18-30 y) and 114 older nonbald men (aged 50-70 y) for a case control comparison. No significant differences were found between cases and controls in allele, genotype, or haplotype frequencies for restriction fragment length polymorphisms of either gene. These findings suggest that the genes encoding the two 5alpha-reductase isoenzymes are not associated with male pattern baldness. Finally, no clear inheritance pattern of male pattern baldness was observed. The relatively strong concordance for baldness between fathers and sons in this study was not consistent with a simple Mendelian autosomal dominant inheritance. A polygenic etiology should be considered.

3-Oxo-5-alpha-Steroid 4-Dehydrogenase↗
PubMed1998

Characterization of novel circovirus DNAs associated with wasting syndromes in pigs.

Porcine circovirus (PCV) was initially recognized as a contaminant of continuous pig kidney cell lines and was not thought to be pathogenic. Antibodies reactive to the cell culture isolate of PCV (PCV PK-15) are prevalent in the swine population worldwide. Recently, PCV PK-15-like antigen and nucleic acid were demonstrated in lesions associated with wasting syndromes in pigs in North America and Europe. Monoclonal antibodies raised to circoviruses isolated from pigs with wasting syndromes highlighted differences between these circoviruses and the PCV PK-15 cell culture isolate. This has led to speculation that a new pathogenic PCV may have emerged in the swine populations of several countries. We report the cloning and characterization of novel circovirus DNAs purified from virus isolates made from tissues of North American and European pigs with wasting syndromes. These North American and European circoviruses form a closely related group at the nucleotide sequence level (> 96% intra-group nucleotide sequence identity) but exhibit < 80% nucleotide sequence identity with the PCV PK-15 cell culture isolate. This report provides evidence for a new type of possibly pathogenic PCV. We propose that these new circoviruses should be referred to as PCV2 as opposed to the original PK-15 cell culture isolate, which should be referred to as PCV1.

Amino Acid Sequence↗
PubMed1998

Effect of moderate exercise on the severity of clinical signs associated with influenza virus infection in horses.

The purpose of this experiment was to determine if exercising horses, infected with influenza virus, exacerbates the severity of clinical disease. Eight horses were trained on a treadmill for 42 days and then challenged with aerosolised influenza A/equine/Kentucky/91 (H3N8). Following challenge, 4 horses (exercise group) continued training for 28 days, while the other 4 horses (nonexercise group) were confined to their stalls. All horses developed clinical signs within 36 h of challenge (fever, coughing, and mucopurulent nasal discharge) and clinical scores were greater in the exercise group. Horses developed fever from Days 1-11 post challenge (PC) and were tachypnoeic and tachycardic from Days 1-14 PC. All horses lost weight within 4 days PC, but the exercise group lost an average of 20 kg more than the nonexercise group. All horses developed pneumonia, and ultrasonography revealed pulmonary consolidation and oedema by Day 7 PC that was resolving by Day 14 PC. Endoscopy and transtracheal aspirates showed airway inflammation for up to 21 days PC. While the exercise group exhibited more severe signs of clinical disease, resolution occurred for both groups on approximately Day 14 PC, and no adverse effects were noted at the end of the study. However, the potential long term effects of exercising horses acutely infected with influenza virus are unknown. Until further research is conducted in this area, it appears prudent not to exercise affected horses.

Analysis of Variance↗
PubMed1998

Isolation of porcine circovirus-like viruses from pigs with a wasting disease in the USA and Europe.

Samples of lung, liver, kidney, pancreas, spleen, and lymph node from pigs with postweaning multisystemic wasting syndrome from California (USA) and samples of mesenteric lymph nodes from similarly diseased pigs from Brittany (France) were examined by light microscopy, in situ hybridization (ISH), and/or virus isolation. Whole genomic probes for porcine circovirus (PCV) and chicken anemia virus (CAV) were used for ISH. Tissue homogenate supernatants were inoculated onto PK/15 cells for virus isolation, and the presence of viral antigen and viral particles was verified by indirect immunofluorescence, ISH, and electron microscopy. Histologic examination of lung from pigs from California revealed interstitial pneumonia, alveolar epithelial hyperplasia, and basophilic nuclear and cytoplasmic inclusions in mononuclear cell infiltrates and various pulmonary epithelial cells. Granulomatous lymphadenitis with syncytial cells typified the lesions seen in the pigs from France. PCV-like nucleic acid was detected by ISH in lung, pancreas, lymph node, kidney, and liver in pigs from California. Positive signal was also obtained in lymph node sections from pigs from France. Probes for CAV were consistently negative. PK/15 cell cultures inoculated with lung preparations from diseased California pigs and mesenteric lymph node preparations from pigs from France had positive fluorescence by indirect staining for PCV using pooled polyclonal pig sera and hyperimmune rabbit serum and had variable staining with a panel of 7 monoclonal antibodies specific for cell culture contaminant PCV. PCV-like nucleic acid was also detected by ISH in cell cultures. Cytopathic effect was not observed. Electron microscopic examination of inoculated cell cultures revealed 17-nm viral particles morphologically consistent with PCV. No other virus particles were observed. Although genomic analysis for the definitive identification of these viral isolates remains to be done, the evidence provided strongly suggests that these tissue isolates are closely related to, although antigenically distinct from, the original PCV cell culture contaminant.

Animals↗
PubMed1998

Aberrant intracellular targeting and cell cycle-dependent phosphorylation of emerin contribute to the Emery-Dreifuss muscular dystrophy phenotype.

The product of the X-linked Emery-Dreifuss muscular dystrophy gene is a protein called emerin, which is localized to the nuclear membrane. We have expressed full-length recombinant human emerin in an in vitro coupled reticulocyte system; it has a molecular mass of 34 kDa, inserts into microsomes in a type II orientation, and does not exhibit any N-linked glycosylation or cleavage event. Affinity-purified human emerin antiserum cross-reacts with the in vitro-expressed emerin and with a 34 kDa band present in a wide range of human tissue samples. Expression and subcellular distribution of emerin were studied in lymphoblastoid cell lines established from four patients with Emery-Dreifuss muscular dystrophy containing different mutations in the emerin gene. Emerin protein was detected in two of these patients by immunoblotting. In striking contrast to wild-type emerin, which was localized to the nuclear fraction and was insoluble in non-ionic detergents and high salt, emerin from these two patients exhibited a more random subcellular localization and increased solubility. On the basis of the mutations present in these patients, it would appear that emerin possesses two non-overlapping nuclear envelope targeting sequences. We have also demonstrated that emerin can occur in four different phosphorylated forms, three of which appear to be associated with the cell cycle. The mutant forms of emerin taken from the two patients exhibited aberrant cell cycle-dependent phosphorylated forms. This data suggests that for emerin to function normally it must be correctly localized, retained at the nuclear membrane and phosphorylated by cell cycle-mediated events.

Adult↗
PubMed1998

Lesions and distribution of viral antigen following an experimental infection of young seronegative calves with virulent bovine virus diarrhea virus-type II.

During the past several years, acute infections with bovine viral diarrhea virus (BVDV) have been causally linked to hemorrhagic and acute mucosal disease-like syndromes with high mortality. The majority of BVDVs isolated in such cases have been classified as type II on the basis of genetic and antigenic characteristics. It was our objective to examine clinical disease, lesions and potential sites of viral replication, following experimental BVDV type II infection in young calves. On approximately day 35 after birth, calves that had received BVDV-antibody-negative colostrum were infected by intranasal inoculation of 5 x 10(5) TCID50 of BVDV type II isolate 24,515 in 5 mL of tissue culture fluid (2.5 mL/nostril). Calves were monitored twice daily for signs of clinical disease. Approximately 48-72 h after infection, all calves developed transient pyrexia (39.4-40.5 degrees C) and leukopenia. Beginning on approximately day 7 after infection, all calves developed watery diarrhea, pyrexia (40.5-41.6 degrees C), marked leukopenia (> or = 75% drop from preinoculation values), variable thrombocytopenia, and moderate to severe depression. Calves were euthanized on days 10, 11, or 12 after infection due to severe disease. Gross and histological lesions consisted of multifocal bronchointerstitial pneumonia (involving 10%-25% of affected lungs), bone marrow hypoplasia and necrosis, and minimal erosive lesions in the alimentary tract. Immunohistochemical staining for BVDV revealed widespread viral antigen usually within epithelial cells, smooth muscle cells and mononuclear phagocytes in multiple organs, including lung, Peyer's patches, gastric mucosa, thymus, adrenal gland, spleen, lymph nodes, bone marrow, and skin. This BVDV type II isolate caused rapidly progressive, severe multisystemic disease in seronegative calves that was associated with widespread distribution of viral antigen and few gross or histological inflammatory lesions.

Animals↗
PubMed1998

A comparison of diagnostic methods for the detection of bovine respiratory syncytial virus in experimental clinical specimens.

Virus shedding was monitored in nasal secretions of 12 calves experimentally infected with bovine respiratory syncytial virus (BRSV) using an antigen capture enzyme-linked immunosorbent assay (ELISA) detecting the nucleoprotein (NP) antigen of BRSV, by a polymerase chain reaction (PCR) amplifying the fusion protein of BRSV, and by a microisolation assay combined with immunoperoxidase staining for the F protein of BRSV. Under the conditions of this study, similar limits of detection and quantitative results were obtained from all three assays. BRSV was detected in nasal secretions of all calves for a minimum of 4 d. Virus shedding began on Day 2 after infection, peaked on Days 3-5, and was cleared in most calves by Day 8. The PCR, and to a lesser extent the ELISA, may detect virus shedding for a longer period after infection than virus isolation, possibly due to neutralization of the virus by rising mucosal antibody. Simulated environmental conditions likely to be experienced during transport of clinical field specimens markedly reduced the sensitivity of virus isolation but had a minimal effect on the results of the NP ELISA. Actual field transport conditions (overnight on ice) had minimal apparent effect on the results of the PCR assay. The less stringent specimen handling requirements, combined with low limits of detection, of both the nucleoprotein ELISA and PCR, indicate either of these assays are more suitable for diagnostic applications than virus isolation.

Animals↗
PubMed1997

Nitric oxide production following in vitro stimulation of ovine pulmonary alveolar macrophages.

Production of inducible nitric oxide (NO) as measured by nitrite in supernatant from ovine pulmonary alveolar macrophage (PAM) cultures was assessed following stimulation of PAM with live cells and supernatants from Corynebacterium pseudotuberculosis and Pasteurella haemolytica cultures; purified bacterial lipopolysaccharide derived from both Escherichia coli and Pasteurella haemolytica alone and in combination with interferon-gamma or lymphocyte conditioned medium; or ovine lentivirus. PAM cultured ex vivo with no further stimulation for 24 h, 48 h or 72 h, produced low concentrations of NO that was not substantially increased following co-culture by the various additives. Assessment of NO production in PAM cultures containing P. haemolytica or supernatant from P. haemolytica cultures was complicated by production of high levels of nitrite in the bacterial cultures. Species differences in inducible NO production may affect the efficacy of clearance of bacterial infections and be responsible for inter-host differences in disease expression following infection by intracellular pathogens.

Animals↗
PubMed1997

Cell-mediated cytolysis of equine herpesvirus-infected cells by leukocytes from young vaccinated horses.

The objective of this study was to determine whether the administration of modified-live equine herpesvirus (EHV-1) to young horses with residual maternal antibodies stimulated EHV-specific cytolytic responses, and whether these responses were crossreactive between EHV-1 and EHV-4. Eighteen clinically normal Belgian cross-foals were used in the study and were commingled in two adjacent pens. Skin biopsies were harvested from 16 foals within 24 h of birth and fibroblast cultures were established, expanded and cryopreserved. Beginning at approximately 10 weeks of age, 10 randomly chosen foals were inoculated on days 0, 21, and 43 of the study with a vaccine containing modified-live EHV-1. Blood mononuclear leukocytes were obtained on days 0, 32, and 50 for the assessment of EHV-specific cytolytic activity using 5 h and 18 h chromium release assays. EHV-1-specific antibodies were assessed by enzyme-linked immunosorbent assay using serum collected on days -21, 0, 32, and 50 of the study. Lymphocyte blastogenic tests and bioassays for interferon activity were conducted on day 50. After two vaccinations, mononuclear leukocytes from seven of ten vaccinated foals had cytolytic activity against autologous EHV-1 cells and leukocytes from six of ten lysed EHV-4-infected cells when tested in an 18 h assay. This activity was enhanced by exogenous interleukin 2 and was markedly reduced using target cells from unrelated horses. Cytotoxicity was not detected in a 5 h assay following in vitro stimulation of leukocytes. After three vaccinations, blood leukocytes from 6/6 vaccinated foals and 0/6 unvaccinated foals had proliferative responses EHV-1. There were no significant differences in interferon production by leukocytes from these foals. Twelve foals tested had low concentrations of (maternal) EHV-1-specific antibody prior to vaccination. Five of eight foals tested had increases in EHV-specific antibodies, while 4/4 commingled unvaccinated foals had a decrease or no change in EHV-specific antibodies. These results demonstrate cytotoxic cellular immune responses can be induced in young horses with maternal antibodies following administration of modified-live vaccine.

Animals↗
PubMed1997

Bovine respiratory syncytial virus.

Since the first report of BRSV in the 1970s, the understanding of this agent and its respective disease has increased dramatically. Current evidence supports a major role for this virus in bovine respiratory disease. Advances in diagnostics have increased the ability to demonstrate this virus in field outbreaks of respiratory disease. The clinical signs and pathologic features have been well described, and vaccines are available to aid in prevention and control. Still, many questions remain to be answered with respect to BRSV. It appears there may be antigenic subgroups of BRSV, but the epidemiologic significance and relevance to immunization of this remains unknown. The question of differences in virulence among isolates of this virus has yet to be addressed. From an epidemiologic standpoint, the means by which BRSV perpetuates in the cattle population has yet to be elucidated. Although progress has been made in understanding the pathogenesis and immune response to BRSV, the mechanism of disease production and immune protection is incomplete. Lastly, efficacy testing of existing vaccines need to continue, as well as the development of new vaccines and new approaches to vaccination.

Animals↗
PubMed1997

Bluetongue virus in laboratory-reared Culicoides variipennis sonorensis: applications of dot-blot, ELISA, and immunoelectron microscopy.

An avidin-biotin complex (ABC) dot-blot, an antigen capture enzyme-linked immunosorbent assay (ELISA), and immunoelectron microscopy (IEM) were used to detect bluetongue (BLU) virus or viral antigen or both in adult Culicoides variipennis sonorensis Wirth & Jones. The dot-blot and ELISA procedures detected viral antigen in 10-22% (depending on serotype) of the biting midges infected with BLU-2, BLU-10, BLU-13, and BLU-17 and approximately 68% of the midges infected with BLU-11. IEM analyses revealed BLU virus in salivary glands, fat body, and thoracic muscle tissue from infected insects. There appeared to be selective growth of the virus in salivary gland tissue.

Animals↗
PubMed1997

Role of tumor necrosis factor-alpha in the ovulatory mechanism of ewes.

Tumor necrosis factor (TNF)-alpha was localized by indirect immunofluorescence microscopy to thecal endothelial cells or preovulatory ovine follicles. Immunostaining of endothelial cells within the apical hemisphere of the follicle declined abruptly with the approach of ovulation; cells within the counterpart basal follicular wall remained largely unaffected. It appeared that TNF alpha had been released into the presumptive area of follicular rupture. Intrafollicular injection of TNF alpha antiserum blocked ovulation in ewes. We considered the possibility that TNF alpha liberated from apical endothelial cells mediates the programmed deletion of ovarian surface and follicular cells that occurs within the formative ovulatory stigma of sheep. Indeed, contiguous ovarian surface epithelium, thecal endothelium, and granulosal cells were protected from apoptotic death (DNA fragmentation) by exposure to TNF alpha antibodies. Finally, TNF alpha induced ovarian cell apoptosis in vitro. We suggest that a bioactive, soluble form of TNF alpha generated from endothelial cells of preovulatory ovine follicles has an obligatory role in the biomechanics of apical follicular weakening and ovarian rupture.

Animals↗
PubMed1997

Performance, survival, necropsy, and virological findings from calves persistently infected with the bovine viral diarrhea virus originating from a single Saskatchewan beef herd.

Fifty-one calves from 652 cows and heifers that calved on a Saskatchewan ranch in 1992 were identified as persistently infected with bovine viral diarrhea virus (BVDV), based on virological and necropsy findings. Herd records suggested a further 20 calves that died between birth and weaning were probably also persistently infected. Subsequent to weaning, all surviving persistently infected calves were transferred to one pen in a 10,000 head commercial feedlot, to mimic normal management practice in western Canadian beef herds. On average, when compared with healthy, BVDV-negative herdmates, persistently infected calves were "poor doers" and had poor survivability, with only 4 persistently infected calves surviving to 1 year of age. There was no difference (P > 0.05) in survival between male and female persistently infected calves. The clinical, pathological, and virological findings from these persistently infected calves varied over time. The majority of persistently infected calves had gross pathological lesions at necropsy, consistent with mucosal disease. However, approximately 25% of the persistently infected calves had gross pneumonic lesions at necropsy, with no or only mild lesions of mucosal disease. A wide variety of other lesions were also noted in persistently infected calves at necropsy. Therefore, the possibility that BVDV-induced lesions can be misdiagnosed is very real. The results of this study indicate that persistent infection with BVDV should always be considered in calves with chronic ill thrift, chronic enteritis, or respiratory disease.

Animals↗
PubMed1997

Systemic adverse reactions in young Simmental calves following administration of a combination vaccine.

Combination vaccines containing viral and bacterial antigens are commonly used in veterinary practice and have been associated with adverse reactions. A group of young Simmental calves developed fever and depression following administration of a mixed vaccine, and 1 died with pulmonary edema, suggesting that endotoxins or other bacterial components may interact synergistically with some adjuvants to cause an enhanced pathologic inflammatory response in some individuals.

Animals↗
PubMed1996

Use of immunohistochemistry and polymerase chain reaction for detection of oncornaviruses in formalin-fixed, paraffin-embedded fibrosarcomas from cats.

OBJECTIVE: To determine whether there was intralesional infection or expression of FeLV or feline sarcoma virus in suspected vaccine-associated fibrosarcomas in cats. DESIGN: Prospective case series. SAMPLE POPULATION: 130 suspected vaccine-associated fibrosarcomas from cats and 1 multicentric fibrosarcoma from 1 cat. PROCEDURE: Excisional biopsy specimens were fixed in formalin and embedded in paraffin. Expression of FeLV antigen was assessed, using a polyclonal goat anti-FeLV glycoprotein 70 (gp 70) serum and an avidinbiotin immunoperoxidase staining technique. The FeLV genome was detected with a polymerase chain reaction (PCR), using primers targeted to a conserved sequence in the untranslated region of the long terminal repeat (LTR) of the FeLV. RESULTS: FeLV gp 70 and LTR sequence were detected in a multicentric fibrosarcoma. All 130 of the suspected vaccine-associated fibrosarcomas were FeLV gp 70 negative on the basis of immunohistochemical test results: 100 fibrosarcomas also were examined by use of PCR and were negative for FeLV LTR region. CLINICAL IMPLICATIONS: Exogenous retroviruses, FeLV, and feline sarcoma virus were not detected in these suspected vaccine-associated fibrosarcomas, using immunohistochemistry and PCR. Additional testing will be required to determine the nature of genomic alterations that are involved in the oncogenesis of vaccine-associated fibrosarcomas in cats.

Animals↗