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Biomedical subjects

J Thorsen

Publications and source records attributed to J Thorsen.

At least 19 recordsLinked to original sources

A case of anaphylactic shock possibly caused by intravesical Hexvix.

BACKGROUND: We report a case of a 69-year-old Caucasian male who experienced a severe drop in blood pressure and a skin rash after fluorescence cystoscopy and transurethral resection of the prostate. The diagnostic agent used for fluorescence cystoscopy and suspected of causing the anaphylaxis was hexaminolevulinate hydrochloride (Hexvix). INCIDENT: A profound fall in blood pressure associated with atrial fibrillation, the appearance of a rash, vomiting and transient loss of consciousness occurred immediately after surgery. The hypotension persisted for several hours despite intensive treatment. The reaction commenced approximately 5 h after exposure to Hexvix. There was an increase in serum tryptase (almost nine-fold) and a positive skin prick test to undiluted Hexvix. CONCLUSION: The mechanism of the anaphylaxis is uncertain, but is considered likely to be a non-immunoglobulin E-mediated allergic reaction possibly caused by Hexvix. This is the first reported case of a severe allergic reaction after intravesical instillation of Hexvix.

Aged↗

High-resolution integrated map encompassing the breast cancer loss of heterozygosity region on human chromosome 16q22.1.

Loss of heterozygosity (LOH) on the long arm of human chromosome 16 is a common genetic alteration observed in both invasive ductal and invasive lobular breast carcinomas. We have generated a high-resolution integrated map encompassing the smallest region of LOH overlap within chromosome 16q22.1 (SRO2). Southern hybridization experiments using more than 140 probes resulted in the assembly of 152 bacterial large-insert clones into a 2.8-Mb contig covering SRO2. The structure of the contig was verified by long-range mapping using total human genomic DNA, and the contig orientation was determined by fluorescence in situ hybridization. A total of 68 transcripts have been identified in the map. One of the genes residing within SRO2 is the E-cadherin gene, CDH1, which has previously been shown to be mutated in lobular breast carcinomas, resulting in loss of E-cadherin expression. In most cases of ductal carcinoma, which is the major mammary cancer type, E-cadherin is normally expressed, suggesting that other genes within 16q22.1 are involved in the development of this tumor subtype. The high-resolution map presented in this study provides a valuable resource for identification of tumor suppressor genes expected to be involved in the etiology of breast carcinomas.

Breast Neoplasms↗

lacZ-neoR transfected glioma cells in syngeneic rats: growth pattern and characterization of the host immune response against cells transplanted inside and outside the CNS.

The rat glioma cell lines BT4C and BT4Cn were stably transfected with the bacterial lacZ-neomycin resistance (neoR) gene construct. Both transfected (BT4ClacZ and BT4CnlacZ) and untransfected cell lines were injected intracerebrally and subcutaneously into rats. Survival time, morphology, growth rate and immunological properties of the tumors were studied. Survival time was significantly prolonged after intracerebral implantation of the transfected cell lines. No similar response was found in nude rats, indicating an immunological response towards the lacZ-neoR-transfected cells in immunocompetent animals. Morphological observations showed that the lacZ-neoR-transfected gliomas were smaller and had a distinct boundary with the normal brain tissue, whereas the parental cell lines revealed a more diffuse growth pattern. Immunostaining showed a higher proportion of immunocompetent cells infiltrating the lacZ-neoR-transfected tumors. After s.c. injection, the lacZ-neoR-transfected BT4C cell line had a prolonged lag phase before assuming a growth rate similar to that of the parental cells. The BT4CnvlacZ tumors initially grew fastest, but then disappeared within 3 weeks. A similar response was observed with mock-transfected tumor cells. A (3)HTdR-incorporation assay on spleen cells from rats transplanted s.c. with BT4CnvlacZ cells showed a 10-fold increase in cell activation as compared with rats with BT4Cn tumors. A humoral response towards the transfected cells was verified by Western-blot analyses.

Animals↗

Infectious agents in acute respiratory disease in horses in Ontario.

A study of acute respiratory disease in horses in Ontario was undertaken to determine the identity of current causative infectious agents. A nasopharyngeal swab was designed and utilized to maximize isolation of viruses, mycoplasma, and pathogenic bacteria. Serum samples were collected for parallel determination of antibody titers to equine influenza virus type A subtype 1 (H7N7) and subtype 2 (H3N8), equine rhinovirus types 1 and 2, equine herpesvirus type 1, Mycoplasma equirhinius, and Mycoplasma felis. Equine rhinovirus type 2 was recovered from 28/92 horses tested, and equine influenza virus type A, subtype 2, was recovered from 5. The mycoplasma and bacteria isolated were consistent with those commonly associated with nonspecific respiratory diseases in horses, except that Streptococcus pneumoniae capsular type 3 was isolated from 10 horses.

Acute Disease↗

Migratory patterns of lac-z transfected human glioma cells in the rat brain.

Malignant brain tumors are characterized by extensive tumor-cell infiltration into the normal brain tissue. The present work describes the migratory behavior of human glioma cells transplanted into the adult rat brain with the aim of exploiting the extent of active cell migration and passive cell displacement within the central nervous system. To detect every transplanted tumor cell, a stably bacterial beta-galactosidase (lac-z) transfected human glioma cell line was used. To distinguish between an active cell migration process and passive cell displacement, rat brains were also implanted with inert fluorescent polystyrene microspheres and the distribution of tumor cells and microspheres was studied 1 hr and 3 days after implantation. One hour after implantation the tumor cells were strictly localized at the implantation site. However, 3 days after implantation, both tumor cells and microspheres showed an extensive distribution within the brain. Confirming earlier neuropathological and experimental studies, it is shown that the lac-z-transfected glioma cells had the capacity to move within the Virchow-Robin and subarachnoid spaces. However, since fluorescent microspheres were also found in these areas, this spread of tumor cells may be primarily mediated by the extensive cerebrospinal fluid flow that exists within the brain. Three days after implantation, the glioma cells also showed an active migration over the corpus callosum. In comparison, the fluorescent microspheres showed only limited spread along the callosal body. It is concluded that the bacterial lac-z gene can be stably transfected into human glioma cells and, since every tumor cell can be visualized within the brain, this model provides a tool for studying the mechanisms behind tumor-cell invasion of the brain.

Animals↗

Chemiluminescent detection of caprine arthritis encephalitis virus with a PCR-generated single stranded nonradiolabelled probe.

A 814-bp digoxigenin-labelled single stranded DNA probe was produced and utilized in slot-blot hybridization for detection of caprine arthritis encephalitis virus (CAEV) in goat synovial membrane (GSM) cell culture infected with CAEV. The sensitivity of a PCR-generated probe was compared with a random primer labelled probe. The probe with digoxigenin-dUTP incorporated in the PCR reaction mixture was more sensitive for RNA detection than the random primer probe and it was much simpler to use. The probe was applied for detection of CAEV by blot blot hybridization in peripheral blood mononuclear cells (PBMC) and macrophage cultures obtained from naturally infected goats. This technique was not sufficiently sensitive to detect the viral nucleic acid directly from PBMC or cultured macrophages. When macrophages were cultured in vitro and then cocultured with susceptible GSM cells, samples gave a positive signal in the slot-blot hybridization technique. The use of slot-blot RNA hybridization permits more convenient and rapid confirmation of CAEV isolation in susceptible cells than the conventional identification by syncytia formation.

Animals↗

Serologic diagnosis of caprine arthritis-encephalitis by ELISA with two recombinant proteins in a parallel testing format.

A scheme for screening sera for antibodies to caprine arthritis-encephalitis virus (CAEV) was evaluated for its ability to identify positive and negative samples in a population with heterogeneous risk factors, using the criteria of sensitivity, specificity and positive predictive value. Five hundred caprine serum samples were tested using a transmembrane recombinant-based ELISA. Those that gave positive results were considered positive, while those with equivocal or negative results were retested with a core recombinant protein-based ELISA. Equivocal results after the second test were considered indeterminate and retesting is advised. Using this approach, a sensitivity, specificity and positive predictive value of 98.8%, 97.2% and 98.6% were obtained. These values are superior to those obtained by these tests used individually. The high sensitivity, specificity and predictive value of this new scheme of CAEV screening make it an attractive addition to any control or eradication program.

Animals↗

Bacterial expression of the caprine arthritis-encephalitis virus gag and env proteins and their use in enzyme-linked immunosorbent assay.

The core protein and the transmembrane protein, encoded for the structural genes gag and env, respectively, of caprine arthritis-encephalitis virus were amplified by use of polymerase chain reaction, cloned into a pGEX-2T vector, and expressed in Escherichia coli as fusion proteins with the glutathione S-transferase at their C-terminus. The recombinant proteins were purified and evaluated by use of an ELISA. Sera from 269 goats were tested, and the results were compared with those obtained by use of immunoblot analysis. When results from both recombinant ELISA (r-ELISA) were compared, it appeared that the transmembrane glycoprotein was more immunoreactive than the core protein, because it was recognized by a higher percentage of sera from infected goats. When results of the 2 ELISA (p28 r-ELISA and p40 r-ELISA) were combined in parallel, they were comparable to those of the immunoblot test, with sensitivity of 100% and specificity of 98.3%. It was also found that use of both r-ELISA makes it possible to compare the variable immunoreactivity against gag and env viral antigens, which may be correlated with the disease state. The r-ELISA, using core and transmembrane proteins, appears to be highly sensitive and specific for detection of antibodies against caprine arthritis-encephalitis virus.

Animals↗

The seroprevalence of maedi-visna in Ontario sheep flocks and its relationship to flock demographics and management practices.

The objectives of this study were to describe the serological prevalence of maedi-visna in a sample of Ontario sheep flocks, and to identify management and demographic variables that were associated with seroprevalence for maedi-visna. A sample of 103 sheep flocks in Ontario was randomly selected from those flocks participating in the Red Meat Plan. The owners of these flocks were surveyed regarding management procedures on their farms, and blood samples were taken from a random sample of ewes in each flock. At least one ewe tested serologically positive, based on the agar gel immuno-diffusion test, in 69.9% of the farms. Positive serological reactions occurred in 20.9% of the 3880 sheep tested. Flock demographics and farm management variables were considered in a multiple regression model, and several factors were positively associated with higher maedi-visna seroprevalence rates. These included the average age of the flock, the number of years the owner had been sheep farming, the practice of using foster ewes, the practice of allowing lambs to have contact with other ewes that are lambing, and the average pasture acreage per ewe.

Animal Husbandry↗

Phocine distemper in a harp seal (Phoca groenlandica) from the Gulf of St. Lawrence, Canada.

The first case of phocine distemper in a seal from Canadian waters and the first case of clinical phocine distemper in a harp seal, Phoca groenlandica, is reported. A two-month-old female harp seal stranded on Prince Edward Island in May 1991. Significant clinical findings were lethargy and severe conjunctivitis. Pulmonary congestion was the main necropsy finding, and histological lesions included diffuse demyelinating nonsuppurative encephalitis and mild multifocal interstitial pneumonia. Acidophilic intracytoplasmic and intranuclear inclusions were present in cerebral neurons and astrocytes. Immunoperoxidase staining confirmed phocine distemper virus (PDV) antigen in the cytoplasm and nuclei of neurons, bronchiolar gland epithelium and transitional epithelium of the bladder. Infectivity titers of canine distemper virus (CDV) (Onderstpoort strain) and a morbillivirus isolated from a grey seal were significantly reduced by serum from the harp seal.

Animals↗

Association of microbiologic flora with clinical, endoscopic, and pulmonary cytologic findings in foals with distal respiratory tract infection.

Undifferentiated distal respiratory tract disease (nasal discharge, cough, pneumonia) in foals (1 to 8 months old) is a burdensome economic problem on breeding farms; yet, the infective agents associated with these episodes have not been well described. Possible causes of these episodes of illness were investigated by culturing specimens of proximal and distal airways of clinically diseased foals (n = 101), prior to any treatment, for aerobic and anaerobic bacteria and viruses (rhinoviruses, equine arteritis virus, equine herpesvirus subtype 1 [EHV-1], influenza virus, and adenovirus). Pairs of sera (n = 47) were examined for antibodies to influenza A virus, equine subtypes 1 and 2, EHV-1, and adenovirus antigens, and sera obtained from foals during acute infection were examined for antibodies (by agar gel immunodiffusion [AGID]) to equi factor antigens of Rhodococcus equi. Viruses were not isolated from the proximal (swab) or distal (bronchial lavage) airway specimens in foals, and only 2 of 47 randomly selected foals seroconverted to EHV-1. Serotiters to the other viruses were low and frequently decreasing between samples, which was compatible with maternally derived antibody. Streptococcus zooepidemicus was the predominant isolate from bronchial lavage specimens (88/101 cases), accompanied by alpha-hemolytic streptococci (8 cases), Bordetella bronchiseptica (13 cases), Staphylococcus epidermidis (9 cases), and other organisms in lesser frequency. Only Str zooepidemicus was recovered significantly (P < 0.05) more often in cases than in controls. The AGID test was found useful to detect foals with presumed exposure to R equi, but positive tests results did not correspond well with bacterial culture results; positive AGID results were recorded in 34% of culture-negative foals.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Effects of hyperoxia on human endothelial cell proliferation and morphology in vitro.

The effects of hyperoxia on endothelial cells from human umbilical veins were studied in vitro by flow cytometric DNA analysis, cell proliferation, and morphology. Exposure of the cells to PO2 of approximately 400 and 500 mmHg (corresponding to O2 concentrations of 65 and 95%, respectively) caused, after 24 h, an increase in the proportion of S-phase cells from 18 to 33%. This was followed by a dramatic decrease of S-phase cells to about 4% after 48 h of O2 exposure. This decrease was also sustained after 72 h of O2 treatment. Concomitant with the depletion of S-phase cells was an increase from 13 to 21% in the proportion of cells in the G2M phase, indicating that cells were unable to undergo cell division after a 48-h exposure to 65 and 95% O2. Cells exposed to various PO2 levels (124, 248, 400, and 500 mmHg) showed a dose-dependent reduction in cell proliferation leading to a total inhibition of cell growth after exposure to PO2 of 400 and 500 mmHg. Cells exposed to 248 mmHg PO2 exhibited similar growth rates as the control cells during the first 3 days. Thereafter there was a 50% reduction in cell density as compared to the control cells. Light and scanning electron microscopy showed that the hyperoxic cells underwent morphologic changes characterized by cell contraction and swelling. Thus it is shown that exposure to very high PO2 tensions first stimulates the cells to enter the S phase of the cell cycle. Thereafter changes take place leading to a nearly complete reduction of cell proliferation.

Cell Count↗

Evaluation of a subunit vaccine for bovine adenovirus type 3.

The hexon subunit of bovine adenovirus serotype 3 (BAV-3) was purified by use of anion-exchange chromatography. A vaccine composed of the purified hexon in immune-stimulating complexes was administered and induced high titer of virus-neutralizing antibody in rabbits and calves.

Adenoviridae↗

An epizootic of Mycoplasma ovipneumoniae infection in captive Dall's sheep (Ovis dalli dalli).

In late spring of 1986, 10 of 23 Dall's sheep (Ovis dalli dalli) at the Metropolitan Toronto Zoo were moved to a new exhibit, where all developed severe respiratory signs refractory to anthelmintic and antibiotic therapy. In July, two animals died with chronic active bronch-pneumonia, and a third was euthanized because of pneumonia several months later. Bacteria were not isolated from the lungs of the first, steptococci and Pasteurella hemolytica were isolated from the other two, respectively; Mycoplasma ovipneumoniae was isolated from both. Pulmonary lesions in all three sheep were consistent with Mycoplasma sp. infection. Nasal swabs of the remaining animals yielded no consistent bacterial isolates; however, four of eight sheep were positive for M. ovipneumoniae. Viral cultures yielded an as yet unidentified herpesvirus. Sheep in the original and new herds had no serologic titers to parainfluenza-3, equine viral rhinopneumonitis, or infectious bovine rhinotracheitis, and had variable titers against bovine respiratory syncytial virus. No titers against M. ovipneumoniae were present in 13 sheep still in the original exhibit, but titers varied from 1:32 to 1:256 in eight pneumonic sheep. Sera taken from three sheep before or early in the outbreak were all negative for antibody to M. ovipneumoniae. Two of the affected Dall's sheep had been in contact with domestic sheep in the winter of 1985-1986, and M. ovipneumoniae was subsequently cultured from the domestic flock. Exposure to a new pathogen, and environmental and social stress in a new exhibit may have resulted in this severe disease in Dall's sheep.

Animals↗

Agents of equine viral encephalomyelitis: correlation of serum and cerebrospinal fluid antibodies.

A survey was conducted by testing 115 paired equine serum and cerebrospinal fluid samples by hemagglutination-inhibition for antibodies to Powassan and snowshoe hare viruses, and by virus neutralization for antibodies to equine herpesvirus type 1. Twenty-five samples were from horses with spontaneous neurological disease and the remainder from horses euthanized because of various nonneurological disorders. All sera and cerebrospinal fluids were negative for antibodies to Powassan virus. Fifty-one sera (44.3%) and 15 cerebrospinal fluids (13.0%) had antibodies to snowshoe hare virus. Ninety-eight sera (85.2%) and four cerebrospinal fluids (3.5%) were positive for antibodies to equine herpesvirus type 1. Powassan virus was inoculated intracerebrally into one, and intravenously into four ponies. Neurological signs associated with a nonsuppurative encephalomyelitis occurred in three ponies. Antibodies to Powassan virus were detected in sera of all animals but in cerebrospinal fluids of only two. Powassan virus was isolated from brain and spinal cord of only the intracerebrally inoculated animal.

Animals↗