Passive immune protection of turkeys against Coryza

Antisera made against whole cells of Bordetella avium protected turkeys against disease signs of turkey coryza, but antiserum against the dermonecrotic heat-labile toxin (DHLT) did not. Neither antiserum against whole cells nor antiserum against DHLT protected turkeys against colonization of the trachea by B. avium. At least 25 bands in whole cell lysate of B. avium separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) reacted in immunoblots with protective antiserum. These bands occurred at < 40 kDa and > 105.3 kDa. DHLT had an isoelectric point (pI) in the range pH 6.3-6.7. Following SDS-PAGE of isoelectric-focused fractions, two bands were recognized by anti-DHLT with immunoblots of pI 6.3, pI 6.5, and pI 6.7 fractions separated by SDS-PAGE.     

Item assessment in the development of a diagnostic motor performance test for myopathy in children

The authors conducted an assessment study of newly developed motor performance items for the diagnosis of myopathy. 17 children who had had muscle biopsy for this diagnosis in the recent past were administered 14 items based on the measurement of static, dynamic and explosive muscle strength and muscle endurance. Individual items did not have perfect discriminative power, but the results were encouraging enough to warrant detailed study with combinations of the items, so that in future fewer children will have to undergo unnecessary muscle biopsies.     

IL-1 in gingival crevicular fluid following closed root planing and papillary flap debridement

Interleukin (IL)-1 alpha and beta are cytokines which can mediate inflammatory, bone resorbing, and reparative effects in the periodontium, but few longitudinal data exist exploring their role following periodontal therapy. This study examined gingival crevicular fluid (GCF) concentrations of IL-1 alpha and IL-1 beta at sites with shallow sulci (SS) or inflamed moderate/advanced pockets (M/AP) before and 6 months after treatment with closed scaling/root planing (SC/RP) or papillary flap debridement (PFD), all in the same subject (n = 14 patients). No significant differences were noted in IL-1 alpha or beta concentrations (determined with two-site enzyme-linked immunosorbent assays) between SS and M/AP sites at baseline. While both therapies improved clinical parameters of periodontal disease, IL-1 alpha concentration increased significantly (p < 0.05) in M/AP-PFD sites 6 months after treatment, but were unchanged in other groups. IL-1 beta concentrations were numerically lower after therapy, except for a significant increase (p < 0.05) in M/AP-PFD sites. These data suggest that surgical wound healing in an inflamed, plaque-infected site (M/AP-PFD) results in prolonged production of IL-1, which may be a reflection of the extent of tissue trauma and delayed wound healing. In spite of increased IL-1 levels, these sites demonstrated significant short-term improvement in clinical attachment level (+ 1.8 mm, p < or = 0.001) postoperatively.     

Murine cytomegalovirus-inhibitory effects of ImuVert

ImuVert, a sterile preparation composed primarily of Serratia marcescens membrane vesicles and ribosomes, was significantly inhibitory to murine cytomegalovirus (MCMV) infections in BALB/c mice. Antiviral activity was manifested as increased survivor number and decreased recoverable virus titers in spleens, lungs and salivary glands. Treatments were intraperitoneal (i.p.) beginning 24 h pre, 4 h post- or 24 h post-virus inoculation and then repeated 4 days later. Doses of 5, 16 or 50 micrograms/mouse were effective; 160 micrograms/mouse, which caused host weight loss in toxicity controls, was not inhibitory to the infection. A single i.p. treatment of mice substantially augmented natural killer (NK) cell activity and increased total B-cells, while reducing total T- and T-helper cells. A late (48 h) decline in T-cell function and transient increases in B-cell function were observed in the treated animals. Serum interferon was not induced. Mice pretreated with anti-asialo GM1 antibody to reduce their NK cell populations, then infected with MCMV and treated with ImuVert were protected to the same degree as normal animals. Severe combined immunodeficient mice infected with MCMV and treated with ImuVert were not protected from the infection. These data suggest ImuVert to act by a mechanism other than NK cell activation in preventing MCMV infections.     

Abnormal hepatic mitochondrial respiration and cytochrome C oxidase activity in rats with long-term copper overload

BACKGROUND: Dietary copper overload in the rat is associated with morphological abnormalities and lipid peroxidation of hepatic mitochondria. This study was designed to determine if copper hepatotoxicity was associated with functional alterations in mitochondrial respiration in conjunction with lipid peroxidation. METHODS: Weanling male rats were pair-fed for 8 weeks on diets containing normal or high levels of copper in combination with sufficient vitamin E. Serum and liver samples were obtained, and hepatic mitochondria were isolated by differential centrifugation. RESULTS: Oxidant injury (decreased levels of hepatic glutathione and alpha tocopherol and increased levels of mitochondrial thiobarbituric acid-reacting substances) was present in the copper-overloaded rats. Serum aminotransferase levels correlated with concentrations of mitochondrial copper and thiobarbituric acid-reacting substances. Copper overload caused a decrease in state 3 respiration and the respiratory control ratio in hepatic mitochondria when several electron donors were used. Analysis of the oxidoreductase activities of the four mitochondrial electron transport protein complexes showed that complex IV (cytochrome C oxidase) activity was reduced by 60% in copper overload. CONCLUSIONS: Functional abnormalities of mitochondria accompany lipid peroxidation and the morphological alterations caused by copper overload, supporting the hypothesis that the mitochondrion is one of the major intracellular targets in copper hepatotoxicity.     

The C-terminus of c-ABL is required for proliferation and viability signaling in a c-ABL/erythropoietin receptor fusion protein

Activated ABL oncogenes cause B-cell leukemias in mice and chronic myelogenous leukemia in humans. However, the mechanism of transformation is complex and not well understood. A method to rapidly and reversibly activate c-ABL was created by fusing the extra-cytoplasmic and transmembrane domain of the erythropoietin (EPO) receptor with c-ABL (EPO R/ABL). When this chimeric receptor was expressed in Ba/F3 cells, the addition of EPO resulted in a dose-dependent activation of c-ABL tyrosine kinase and was strongly antiapoptotic and weakly mitogenic. To evaluate the contributions of various ABL domains to biochemical signaling and biological effects, chimeric receptors were constructed in which the ABL SH3 domain was deleted (triangle upSH3), the SH2 domain was deleted (triangle upSH2), the C-terminal actin-binding domain was deleted (triangle upABD), or kinase activity was eliminated by a point mutation, K290M (KD). The mutant receptors were stably expressed in Ba/F3 cells and analyzed for signaling defects, proliferation, viability, and EPO-induced leukemia in nude mice. When compared with the ability of the full-length EPO R/ABL receptor to induce proliferation and support viability in vitro, the triangle upSH3 mutant was equivalent, the triangle upSH2 mutant was moderately impaired, and the triangle upABD and KD mutants were profoundly impaired. None of these cell lines caused leukemia in mice in the absence of pharmacological doses of EPO. However, in mice treated with EPO (10 U/d), death from leukemia occurred rapidly with wild-type and triangle upSH3. However, time to death was prolonged by at least twofold for triangle upSH2 and greater than threefold for triangle upABD. This inducible model of ABL transformation provides a method to link specific signaling defects with specific biological defects and has shown an important role for the C-terminal actin-binding domain in proliferation and transformation in the context of this receptor/oncogene.     

Role of glutathione, glutathione S-transferases and multidrug resistance-related proteins in cisplatin sensitivity of head and neck cancer cell lines

Spontaneous proliferative liver lesions were found in 15 (13 males and 2 females) of 244 (122 of each sex) transgenic (Tg) mice carrying the human prototype c-H-ras gene (rasH2). The liver lesions included 3 foci of cellular alteration, 1 hepatocellular adenoma, 5 hepatocellular carcinomas, and 4 hepatic hemangiosarcomas in the males and 1 focus of cellular alteration and 1 hepatocellular carcinoma in the females. The mutation patterns of the human and endogenous mouse c-H-ras codon 61 in these proliferative liver lesions were analyzed by DNA amplification using polymerase chain reaction, single-strand conformation polymorphism (PCR-SSCP), and oligonucleotide dot blot hybridization. The hepatocellular carcinomas in 4 males and 1 female contained a point mutation in the mouse c-H-ras gene: 3, 1, and 1 carcinomas had a CAA to AAA transversion at the first base of codon 61, a CAA to CTA transversions, and a CAA to CGA transition at the second base of codon 61, respectively. No point mutations in the human c-H-ras transgene were detected in any hepatocellular carcinoma. All 4 hepatic hemangiosarcomas had a CAG to CTG transversion at codon 61 of the human c-H-ras gene, but no point mutations were detected in codon 61 of the mouse c-H-ras gene. No mutations in human or mouse c-H-ras codon 61 were detected in altered cell foci or hepatocellular adenoma. These results indicate that spontaneous liver tumors in rasH2 Tg mice contain different mutation patterns depending on the histologic type or cell origin of the tumors (i.e., hepatocellular carcinomas or hepatic hemangiosacomas). The absence of similar mutations in foci of cellular alteration and the hepatocellular adenoma suggests that the occurrence of codon 61 point mutations is a late event in the progression of hepatocellular neoplasia in rasH2 Tg mice.