Cases reported "Disease Models, Animal"

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1/14. Usefulness of procalcitonin in Pseudomonas burn wound sepsis model.

    Procalcitonin (PCT), a precursor of calcitonin, and endotoxin were determined in the burn wound sepsis model in which 21 Sprague-Dawley rats were scalded approximately 30% on their back. On day 2 post burn, the wounds were inoculated 1 x 10(8) colony-forming units of pseudomonas aeruginosa. On day 5 post burn P. aeruginosa was detected by blood culture in 10 of the 21 rats (47.6%). The mortality rate 7 days after burn was 90.5%. Significant correlations were observed between serum endotoxin levels and serum PCT levels on day 5 post burn (r = 0.860, p<0.001). It was suggested that endotoxin may induce the release of PCT and that measuring the levels of PCT may be useful in diagnosing burn wound sepsis.
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2/14. Scopulariopsosis and hypersensitivity pneumonitis in an addict.

    Granulomatosis caused by fungal spores of a soil saprophyte is a newly recognized pulmonary complication of intravenous drug addiction. Brown, non-budding spores were histologically identified in necrotic tissue, inside giant cells of sarcoidlike granulomata, and in the vicinity of focal angiitic lesions. The fungus was identified by culture as the dematiaceous scopulariopsis brumptii. Cultural and histopathologic studies of lung biopsy specimens established the diagnosis. We showed precipitating antibodies to fungal antigen in the serum, prepared from the patient's isolate. Similar granulomatous pulmonary lesions were experimentally produced in mice by a single intravenous injection of spores of S. brumptii. The spores remained viable but did not show evidence of growth in the animal's tissue. Precipitating antibodies to fungal antigen and immediate wheal and late necrotizing type of skin reactions were shown in the challenged mice. The studies support the notion that pulmonary hypersensitivity to fungal spores was mediated by an Arthus'-type phenomenon.
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3/14. Pathogenesis of cerebral cryptococcus neoformans infection after fungemia.

    The pathogenesis of cerebral infection after cryptococcus neoformans fungemia in outbred mice was investigated. Confocal microscopy and cultures on ficoll-hypaque gradient-separated blood cells were used to detect yeasts in the cytoplasms of monocytes. In semithin brain sections, poorly capsulated yeasts were seen in macrophages in the leptomeningeal space, in monocytes circulating in leptomeningeal capillaries, or in the endothelial cells themselves, strengthening the hypothesis that monocytes and endothelial cells play key roles in the pathogenesis of cryptococcal meningitis. Similar fungal loads and cellular reactions were seen in mice and in 1 patient with acquired immune deficiency syndrome (AIDS), all with acute cryptococcal meningoencephalitis, and in mice and in 1 patient with AIDS, all with cured cryptococcal infection. Immunostaining revealed both the presence of cryptococcal polysaccharide in various brain cells and antigenic variability both from yeast cell to yeast cell and over time. Thus, our data established the relevance and interest that this experimental model has for investigation of the pathogenesis of human cryptococcal meningitis.
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4/14. Regulation of cutaneous pigmentation by titration of human melanocytes in cultured skin substitutes grafted to athymic mice.

    pigmentation of healed cultured skin substitutes in burn patients is frequently irregular and unpredictable which compromises solar protection and the patient's self-image. To address these morbidities, human fibroblasts were inoculated on a collagen-glycosaminoglycan substrate followed 1 day later by the addition of keratinocytes at 1.1 x 10(6)/cm2 combined with either 0, 1.1 x 10(2), 1.1 x 10(3), or 1.1 x 10(4) melanocytes/cm2. The skin substitutes were incubated in vitro for 3 weeks and grafted to athymic mice. in vitro, the number of L-Dopa-positive melanocytes in the skin substitutes increased proportionately to the number of melanocytes inoculated. The melanocytes localized to the basal epidermis when labeled for MEL-5. The skin substitutes with 1.1 x 10(4) melanocytes/cm2 were significantly darker than other groups in vitro by chromameter evaluation. By 12 weeks after grafting, the cultured skin ranged from no pigment in the control group, to 75% pigmented area in the 1.1 x 10(3) melanocytes/cm2 group, to complete pigmentation in the 1.1 x 10(4) melanocytes/cm2 group. In vivo, the mean chromameter values were significantly darker for the grafts with 1.1 x 10(3) and 1.1 x 10(4) melanocytes/cm2. These results suggest that complete restoration of cutaneous pigmentation can be accomplished by addition of between 0.1 and 1.0 x 10(4) melanocytes/cm2 to skin substitutes.
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5/14. Animal models for lysosomal storage diseases: a new case of feline mucopolysaccharidosis vi.

    Two long-haired Siamese cats are reported with clinical manifestations of human mucopolysaccharidosis vi (Maroteaux-Lamy disease): facial dysmorphia, dysostosis multiplex, paralysis. urine of the two affected animals contained a high concentration of glycosaminoglycans, as detected by the dimethylmethylene blue test. Qualitative analysis, performed by thin-layer chromatography of the cetylpyridinium chloride-precipitable material, showed dermatan sulphate. Excessive incorporation of [35S]sulphate in the intracellular mucopolysaccharide of cultured fibroblasts and deficiency of arylsulphatase B in such cells indicate that these cats are affected by Maroteaux-Lamy disease. They should thus be considered the first European case of feline mucopolysaccharidosis vi.
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6/14. Prospects for human gene therapy.

    Retroviral vectors containing marker genes and the sequence for human proteins have been used to transduce cultured lymphocytes, which have then been reinfused into patients. Circulating hematopoietic progenitor cells from human fetal cord blood obtained at the time of term and premature deliveries as early as 19 weeks of gestation have been shown to express such transduced genes in vitro. Cord blood cells from fetal sheep sampled and transduced ex vivo and transfused back in utero expressed marker genes up to two years after birth. Although the efficiency of gene transfer into cells and their long-term expression need to be improved, the potential exists for treating some genetic diseases after prenatal diagnosis either in utero or shortly after birth.
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7/14. Failure of therapeutic coma and ketamine for therapy of human rabies.

    The recent success in treating a human rabies patient in Milwaukee prompted the use of a similar therapeutic approach in a 33-year-old male Thai patient who was admitted in the early stages of furious rabies. He received therapeutic coma with intravenous diazepam and sodium thiopental to maintain an electroencephalographic burst suppression pattern, which was maintained for a period of 46 h, as well as intravenous ketamine (48 mg/kg/day) as a continuous infusion and ribavirin (48 to 128 mg/kg/day) via a nasogastric tube. He never developed rabies virus antibodies and he died on his 8th hospital day. At least three other patients have been treated unsuccessfully with a similar therapeutic approach. Because of the lack of a clear scientific rationale, high associated costs, and potential complications of therapeutic coma, the authors recommend caution in taking this approach for the therapy of rabies outside the setting of a clinical trial. More experimental work is also needed in cell culture systems and in animal models of rabies in order to develop effective therapy for human rabies.
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8/14. salmonella infection in total hip replacement: tests to predict the outcome of antimicrobial therapy.

    We report a hematogenous implant infection with salmonella dublin in a renal transplant patient with total hip replacement. A 16-month treatment with cotrimoxazole failed, as evidenced by culture and electron microscopy, despite persisting low MIC after therapy. Data from a foreign body animal model and in vitro tests, which take into account the properties of adhering and stationary-phase bacteria, explain the failure of a long-term treatment with cotrimoxazole. The patient was subsequently cured by ciprofloxacin which was successful in these tests. No relapse was noted after a follow-up of 1 year.
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9/14. Clinical and experimental keratitis due to Curvularia lunata (Wakker) Boedijn var. aeria (Batista, Lima and Vasconcelos) Ellis.

    A case of mycotic keratitis due to Curvularia lunata var. aeria is reported for the first time from india. Direct microscopic examination of the corneal scrapings in KOH (50 g1(-1)), stained with lactophenol cotton blue, revealed brown septate hyphae and the cultures were found positive for fungus. It was identified as C. lunata (Wakker) Boedijn var. aeria (Batista, Lima and Vasconcelos) Ellis (IMI 253204) and its growth was better at 28 degrees C than at 37 degrees C. in vitro, the fungus was most sensitive to amphotericin b followed by miconazole nitrate, clotrimazole and triphenyltin isoselenocyanate 4-picoline which had the same activity. lactones were least active and tolciclate and arnebins and inactive. The fungus produced experimental keratomycosis in the rabbit cornea and the infection severity as well as histopathological changes were more pronounced in rabbits that received penicillin, streptomycin and cortisone.
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10/14. Establishment of a human B-CLL xenograft model: utility as a preclinical therapeutic model.

    Chronic lymphocytic leukemia (CLL), a proliferative disease of mature looking B lymphocytes, is the commonest leukemia in western countries. It remains incurable by available treatment modalities. We report on the establishment of a permanent, EBV-negative, B-CLL line (WSU-CLL) from the peripheral blood of a patient with CLL. The cells grow as suspension in liquid culture, express IgG lambda and other B cell markers and show lg heavy and light gene rearrangements. Karyotypic analysis shows 45,X,del(3)(p14;p24),t(4;12;12) (q31;q22;p13), t(5;12) (q31;p13), add(16)(q24)X2, t(18;21) (q12;p12). WSU-CLL forms colonies when grown on soft agar. A xenograft model was established by injecting the WSU-CLL cells subcutaneously (s.c.) in severe combined immune deficient (SCID) mice. When the s.c. tumor was transplanted in vivo to other SCID mice, the success rate was 100% with a doubling time of 7.3 days. The CLL-SCID xenograft model was used to test the efficacy of selected standard chemotherapy drugs and new therapeutic agents against WSU-CLL. The cell line and the xenograft described can be used as a model to facilitate the development of new therapeutic agents against CLL in man.
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