Orthovoltage radiation and weekly low dose of doxorubicin for the treatment of incompletely excised soft-tissue sarcomas in 39 dogs.

The efficacy and toxicity of orthovoltage radiation therapy and concurrent low doses of doxorubicin for the treatment of incompletely excised soft-tissue sarcomas in 39 dogs was investigated retrospectively. The 39 dogs had 40 soft-tissue sarcomas and received 51 Gy orthovoltage radiation in 17 daily 3 Gy fractions; they also received 10 mg/m(2) doxorubicin once a week administered intravenously one hour before the dose of radiation. The median follow-up time was 910 days. The tumours recurred locally in seven of the dogs, in five of them within the radiation field; the median time to their recurrence was 213 days (range 63 to 555 days). Six of the dogs developed a distant metastasis after a median time of 276 days (range eight to 826 days). The one-year and two- to four-year tumour control rates were 84 per cent and 81 per cent, respectively, and the one-, two- and three- to four-year survival rates were 85 per cent, 79 per cent and 72 per cent, respectively. Tumours with a mitotic rate of more than 9 per 10 high-power fields were significantly more likely to recur, and the dogs with such tumours survived for significantly shorter periods.

Family history as a risk factor for ulcerative colitis-associated colon cancer in cotton-top tamarin.

BACKGROUND & AIMS: Little is currently known about the relationship between family history of colon cancer and ulcerative colitis-associated colon cancer. A nested case-control study was performed to evaluate the association between family history of colon cancer and spontaneously occurring colon cancer in cotton-top tamarins (Saguinus oedipus). METHODS: Subjects were chosen from a colony of cotton-top tamarins held in captivity between 1968 and 1995. The cancer status of parents and grandparents was compared for 48 animals with colon cancer and 58 controls, all with histological confirmation of ulcerative colitis. Multivariate odds ratios were calculated using logistic regression. RESULTS: A parental history of colon cancer was positively associated with risk of colon cancer (multivariate odds ratio, 2.7; 95% confidence interval, 1.1-6.3). Risk also increased as an animal's total number of family members with colon cancer increased (multivariate odds ratio, 1.7 for each increase in the total number of family members with cancer; 95% confidence interval, 1.1-2.8). CONCLUSIONS: The results suggest that cotton-top tamarins with ulcerative colitis are at significant increased risk for developing colon cancer if they have a family history of colon cancer. Further investigation of this relationship in both tamarins and humans is warranted.

A prospective study of the epidemiology of colitis and colon cancer in cotton-top tamarins (Saguinus oedipus).

BACKGROUND & AIMS: Spontaneous colitis and colon cancer in the cotton-top tamarin have been shown to resemble human ulcerative colitis and its associated cancer. The effect of environment and diet on the evolution of the disease was studied in animals from birth to 5 years of age. METHODS: Newborn tamarins were assigned to three groups reared in (1) a colony in which colitis was highly prevalent and fed a standard diet; (2) an isolation unit and fed a standard diet or one of two semipurified diets; and (3) a multispecies nursery, returned to the colony, and fed the same semipurified diets. Mucosal biopsy specimens from the descending colon were taken at 4-month intervals. RESULTS: Acute colitis and chronic mucosal changes were significantly higher in the colony than in the isolation unit. Diet had no effect on acute colitis, but chronic mucosal changes were significantly higher in animals fed a standard diet than semipurified diets. CONCLUSIONS: Data suggest that acute colitis was associated with environment. Factors in the environment, including a transmissible agent, are discussed. Chronic mucosal changes were modified by diet. Cancer was associated with acute colitis and chronic changes and seems to be associated with diet.

Spontaneous renal lymphosarcoma in a juvenile cynomolgus monkey (Macaca fascicularis).

Spontaneous lymphosarcoma, likely of renal origin, was diagnosed in a naive, juvenile, male cynomolgus monkey (Macaca fascicularis). Histologically, renal architecture was effaced by dense infiltrating sheets of plump cells with indistinct borders and scant amphophilic cytoplasm. Mitoses were uncommon. Similar neoplastic infiltrates were also present in the right renal cortex, one adrenal medulla, the prostate, seminal vesicles, myocardium, and the pulmonary interstitium. Serological tests were negative for infection with Simian Immunodeficiency Virus (SIV), Simian T-Lymphotropic Virus-1 (STLV-1), and Simian Retrovirus-5.

Disseminated B virus infection in a cynomolgus monkey.

A cynomolgus monkey (Macaca fascicularis) was euthanatized 1 week following dystocia because of severe peritonitis. Histologic examination revealed lesions characteristic of herpesvirus infection in lungs, liver, spleen, bone marrow, uterus, and adrenal gland, and on the serosal surface of intestines, pancreas, and reproductive tract. Immunohistochemical studies on liver and lungs revealed Herpes B-like antigens in the lesions. B virus was isolated from serum. As systemic B-virus infection was not diagnosed before death of the monkey, these findings underscore the need for universal precautions when handling blood, fluids, or tissues from macaques.

Simian virus 40-induced disease in rhesus monkeys with simian acquired immunodeficiency syndrome.

Simian virus 40 (SV40) disease was diagnosed in four rhesus monkeys that died with SIV-induced acquired immunodeficiency syndrome (AIDS). One juvenile monkey seroconverted for SV40 6 months after inoculation with SIV and developed severe bilateral tubulointerstitial nephritis. In contrast, progressive multifocal leukoencephalopathy (PML) occurred in two adult monkeys that were seropositive for SV40 before SIV inoculation, as well as a third adult that was naturally infected with SIV and seropositive for SV40 5 years before death. Large intranuclear inclusions containing abundant polyomavirus particles were limited to either renal tubular epithelial cells or oligodendrocytes. In situ DNA hybridization for SV40 large T antigen further demonstrated that SV40 nucleic acid was localized to either kidney or brain tissue. By immunohistochemical analysis, areas of central nervous system inflammation and demyelination were shown to contain CD68+ macrophages (gitter cells), aggregates of CD8+ T lymphocytes, and numerous gemistocytic astrocytes that labeled for glial fibrillary acidic protein. These observations indicate that rhesus monkeys with SIV-induced AIDS are predisposed to polyomaviral disease, in which SV40 nucleic acid is observed in renal tissue in primary infections and brain tissue after viral reactivation. Furthermore, this organ-specific replication suggests that tissue-tropic strains of SV40 may develop in immunodeficient monkeys.

Ulcerative colitis-linked antineutrophil cytoplasmic antibody in the cotton-top tamarin model of colitis.

The presence of antineutrophil cytoplasmic antibodies (ANCA) in serum has recently been shown to be a good disease marker for ulcerative colitis. An animal model of colitis expressing serum ANCA would allow prospective studies of the relationship between ANCA expression and development of colitis. The cotton-top tamarin model of spontaneous colitis was investigated for presence of ANCA to evaluate its potential as a model for the study of the immune response in human ulcerative colitis. Results show that cotton-top tamarin neutrophils have antigens recognized by immunoglobulin G (IgG) from the ANCA-positive serum of patients with ulcerative colitis. However, cotton-top tamarins do not generate serum IgG reactive to cotton-top tamarin or human neutrophils even when equal amounts of IgG are used for antigen detection. As new animal models of spontaneous and induced chronic colitis are developed, the presence of ANCA in serum, using the species' neutrophils, will be useful in relating the immune response of the model to that in human ulcerative colitis.

Effect of simian immunodeficiency virus infection on tumor necrosis factor-alpha production by alveolar macrophages.

We studied the release of tumor necrosis factor-alpha (TNF alpha), a vital immunoregulatory cytokine, by alveolar macrophages (M phi s) infected with simian immunodeficiency virus (SIV) in vitro or collected from SIV-infected macaques. For in vitro studies, M phi s were harvested by bronchoalveolar lavage from 5 normal animals and infected in flasks with SIV (10(4)TCID50/2.5 x 10(6) M phi s). After 7 to 10 days, cytopathic effect was prominent and 68 +/- 2% of M phi s were immunoreactive for p27 core protein. Uninfected (control) and SIV-infected M phi s were then cultured for 24 hours in 96-well plates (10(5) M phi s/well) while challenged with lipopolysaccharide (LPS; 100 micrograms/ml). TNF alpha was assayed in culture supernatants by an enzyme-linked immunosorbent assay (detection limit, 50 pg/ml) and results were expressed as pg TNF alpha/ml/10(3) M phi s (mean +/- SEM). TNF alpha was not detected in unstimulated wells. TNF alpha release by control and SIV-infected M phi s was similar (6.6 +/- 0.7 and 7.9 +/- 1.1 pg/ml/10(3) M phi s, respectively). We also studied TNF alpha release by alveolar M phi s from 8 animals infected with SIV (3 asymptomatic, 5 with acquired immune deficiency syndrome virus (AIDS]. One animal with AIDS had p27+ M phi s. Alveolar M phi s from asymptomatic animals released significantly more TNF alpha (10.3 +/- 1.1 pg/ml/10(3) M phi s) than did animals with AIDS or uninfected macaques (5.2 +/- 0.8 and 7.0 +/- 0.6 pg/ml/10(3) M phi s, respectively) (p less than 0.01). However, M phi s from monkeys with AIDS failed to respond to LPS after 7 to 10 days in culture. In summary, in vitro infection with SIV does not cause constitutive TNF alpha release or alter the response of cultured M phi s to LPS. When kept in culture, M phi s collected from asymptomatic, SIV-infected animals retain their response to LPS, whereas M phi s from animals with AIDS lose the capacity to produce TNF alpha. Furthermore, M phi s cytokine production is exaggerated before overt clinical disease, but not as a direct result of infection with SIV.

Distinctive unclassified mesenchymal tumor of the digit of dogs.

Four examples of a mesenchymal tumor of undetermined histogenesis occurred in three mixed-breed dogs and one Yorkshire terrier. All tumors occurred as solitary, soft to firm, solid, tan, and ulcerated masses in the digits of dogs aged 11 to 15 years. The compact cellular tumor had cells with anisokaryotic round, oval, or irregular nuclei, some of which were multinucleated. The neoplastic cells appeared to arise in the tissue near the third phalanx in the area of dense collagenous trabeculae located proximal to the fat pad and sweat glands. The unclassifiable cells had some features of histiocytes by transmission electron microscopy, but failed to stain for lysozyme and alpha-1-antichymotrypsin, markers for monocyte-macrophage derived cells. Immunohistochemically, the cells stained for vimentin but not for cytokeratins, desmin, S-100 protein, epithelial membrane antigen, alpha-lactalbumin, lysozyme, alpha-1-antichymotrypsin, alpha-lactalbumin, casein, and heavy and light chain immunoglobulins. The combined findings of light and transmission electron microscopy and immunohistochemistry exclude tumor histogenesis from an epithelial cell, melanocyte, mast cell, plasma cell, Schwann cells, and Merkel cell.

Identification of transforming genes of subgroup A and C strains of Herpesvirus saimiri.

Herpesvirus saimiri is an oncogenic herpesvirus that induces rapidly progressing lymphomas in New World primates. Using retrovirus vectors for gene transfer, specific open reading frames of H. saimiri were tested for their ability to transform rodent cells in culture. One open reading frame, designated STP-C488 (for saimiri-transformation-associated protein of the subgroup C strain 488), phenotypically transformed Rat-1 cells, resulting in formation of foci, growth at reduced serum concentration, and growth to higher cell densities. Cells transformed by STP-C488 formed invasive tumors in nude mice. The STP-A11 reading frame of strain 11 (subgroup A) was much less potent in its transforming ability than STP-C488. These results demonstrate the oncogene nature of these two open reading frames and provide a means for studying their transforming functions independent of the rest of the H. saimiri genome.

Macrophage-tropic variants of SIV are associated with specific AIDS-related lesions but are not essential for the development of AIDS.

The importance of macrophage infection for the development of acquired immune deficiency syndrome (AIDS) was investigated. Molecularly cloned simian immunodeficiency virus (SIV)mac239 replicates very poorly in cultured macrophages yet it causes AIDS in rhesus monkeys. Three of five rhesus monkeys that died with AIDS following SIVmac239 infection showed no disease manifestations directly associated with macrophage infection, such as encephalitis and granulomatous interstitial pneumonia. Simian immunodeficiency virus recovered from the peripheral blood of these three animals at or near the time of death replicated very poorly if at all in cultured macrophages, and tissues taken at autopsy showed little or no infection of macrophages by immunohistochemical staining. However two of the five rhesus monkeys that died with AIDS following SIVmac239 infection displayed a characteristic SIV-related meningoencephalitis and/or granulomatous pneumonia, lesions associated with macrophage infection. Simian immunodeficiency virus recovered from the peripheral blood of these two animals near the time of death replicated extremely well in cultured macrophages, indicating the emergency of macrophage-tropic variants in vivo. Furthermore tissues taken at autopsy from these two showed many infected macrophages by immunohistochemical staining. These results indicate that AIDS and death can occur without obvious involvement of macrophage infection. However the presence of macrophage-tropic viral strains appears to influence the disease course and disease manifestations.

Cutaneous herpesvirus infection in a mallard duck (Anas platyrhynchos).

Investigations of mortalities involving wild mallard ducks (Anas platyrhynchos) revealed the presence of a herpesvirus associated with skin lesions on the plantar surface of the foot web of one duck. Ultrastructurally, the paracrystalline arrays of viral core particles and unencapsidated nucleoids in the nucleus, and the enveloped viruses in cytoplasmic vacuoles are compatible with a herpesvirus. This appears to be the first report of cutaneous lesions in a mallard duck attributable to a herpesvirus. Whether this lesion is due to duck virus enteritis (DVE), suggesting that cutaneous shedding is possible, or due to another uncharacterized herpesvirus of ducks is unclear.

Study of long-term cultures of simian immunodeficiency virus (SIVmac 251)-infected peripheral blood lymphocytes.

A culture of rhesus monkey peripheral blood lymphocytes was divided into two parts; one was kept as an uninfected control, and the other was infected with a strain of simian immunodeficiency virus (SIVmac251) originally isolated from a rhesus monkey that died of a malignant lymphoma associated with acquired immune deficiency syndrome. Both cultures were sampled at successive intervals from 1 to 40 days postinfection. Each sample was subjected to in situ hybridization for detection of viral mRNA, immunocytochemical detection of viral core protein (p27), reverse transcriptase assay, electron microscopy, and immunophenotypic characterization of infected cells. These techniques were used to define viral growth kinetics of this novel lentivirus in peripheral blood lymphocytes. The first evidence of SIVmac251 replication was obtained by an in situ hybridization signal for viral mRNA at 2 days postinoculation. This was followed by detection of viral p27 core protein by immunocytochemistry on day 4. Reverse transcriptase activity above control values was not detected until day 8. Budding particles were not found in the infected cultures until 14 days postinfection. Results of in situ hybridization, immunocytochemistry, and reverse transcriptase assay indicated that two bursts of viral replication occurred during the course of this study. The first, at 3 weeks postinfection, was due to infection and subsequent depletion of CD4+ lymphocytes, while the second, 3 weeks later, resulted from a cycle of replication in CD8+ lymphocytes and the remaining CD4+ cells, culminating in the death of all cells on day 39 postinoculation.

Chordoma in a cat.

A chordoma within the deep musculature adjacent to C3 and C4 was excised from a 14-year-old castrated domestic cat. Metastatic chordoma developed in a prescapular lymph node 10 months later. At necropsy 11 months after complete excision of the primary tumor, metastases were found in both retropharyngeal lymph nodes.

Soluble and membrane-associated interleukin 2 receptor-alpha expression in rhesus monkeys infected with simian immunodeficiency virus.

More than 80% of rhesus monkeys infected with simian immunodeficiency virus (SIV) were found to have elevated levels of soluble interleukin-2 receptor (IL-2R) in their serum during the course of infection. All long-term survivors had stably elevated levels of soluble IL-2R. The highest levels of soluble IL-2R correlated with the expression of IL-2R on tissue macrophages. Although IL-2R expression was induced on alveolar macrophages by infection with SIV in vitro, expression of IL-2R on tissue macrophages in vivo was not associated with concurrent SIV protein expression in the same cells. Moreover, in animals with high soluble IL-2R levels, there was an inverse relationship between the numbers of cells expressing IL-2R and cells expressing viral protein. The results suggest that the induction of IL-2R may be an indirect or secondary effect of SIV infection. Changes in expression of macrophage-elaborated factors, such as that of IL-2R described in this report, may play a crucial role in some of the pathologic features of acquired immunodeficiency syndrome.

Immunologic and pathologic manifestations of the infection of rhesus monkeys with simian immunodeficiency virus of macaques.

The striking similarities between simian immunodeficiency virus (SIV)-induced disease in macaque monkeys and HIV-induced disease in humans make the SIV-induced macaque monkey an extraordinarily important model for the study of AIDS. The most significant difference between these lentivirus-induced syndromes is the more rapid progression of disease in SIV-infected monkeys. The immunologic and pathologic manifestations of SIV infections in rhesus monkeys are described.

Use of simian immunodeficiency virus for vaccine research.

Rhesus monkeys were immunized with purified, disrupted, noninfectious simian immunodeficiency virus (SIV) in adjuvant induced SIV neutralizing antibodies. Two of six previously vaccinated macaques were protected against infection when challenged with 200-1,000 animal infectious doses of uncloned, pathogenic SIV and both have remained free of signs of virus infection for 19 and 30 months. Prior vaccination appeared to be of benefit in decreasing the virus load and in delaying the onset of AIDS in animals that became infected. Nonetheless, two of four previously vaccinated monkeys that became infected following challenge eventually developed AIDS and died 505 and 538 days after infection. Thus, for a vaccine to be truly effective against AIDS, it may have to protect absolutely against initial infection.