Evaluation of maternal bromoform supplementation in late gestation on blood parameters of cows and their progeny.

Bromoform supplementation has been successful in reducing enteric methanogenesis in ruminants; however, the impacts on the health of these animals are still limited. The current study evaluates the impact of maternal bromoform supplementation on the health of late-gestation cows and their progeny. Pregnant Angus cows (n = 42) were allocated into a control or bromoform group (n = 21 cows per treatment). Bromoform extracted from Asparagopsis armata (7,372 mg/kg) was supplemented once daily. Blood samples were collected from cows before supplementation (baseline). Within 24 h of parturition, blood and colostrum samples were collected from each cow and blood from neonates. Colostrum brix was measured to indicate immunoglobulin content. All data was analysed using the MIXED procedure in SAS. Supplementation of cows with bromoform resulted in increased blood urea to creatinine ratio (P = 0.048), base excess (P = 0.049), total carbon dioxide (TCO2; P = 0.048) and a decrease in blood glutamate dehydrogenase (GLDH; P = 0.031) compared to the control group. For cows in the bromoform group, a trend was observed for higher levels of partial pressure of carbon dioxide (pCO2; P = 0.070) and bicarbonate (HCO3-; P = 0.052), and lower levels of partial pressure of oxygen (pO2; P = 0.058) compared to the control group. Blood gamma-glutamyl transferase (GGT) was elevated in offspring of cows fed bromoform (P = 0.050). The lower blood pO2 of pregnant cows fed bromoform and elevated blood GGT levels in offspring are not well understood and highlight the need for further investigation. Additionally, the low-dose bromoform supplementation affected various blood gas parameters of cows and calves, demonstrating the importance of monitoring these parameters when using different doses of halogenated compounds in livestock.

Pathologic findings in weedy (Phyllopteryx taeniolatus) and leafy (Phycodurus eques) seadragons.

A retrospective study of the pathologic findings in weedy (Phyllopteryx taeniolatus) and leafy (Phycodurus eques) seadragons was performed on specimens submitted to 2 reference laboratories from 1994 to 2012 to determine the range and occurrence of diseases affecting aquarium-held populations. One hundred two and 94 total diagnoses were recorded in weedy and leafy seadragons, respectively. Two of the more common etiologic diagnoses in both species were mycobacteriosis and scuticociliatosis, whereas myxozoanosis was common in weedy seadragons. Metazoan parasite infections were less common etiologic diagnoses. There were no correlations between mycobacteriosis and ciliate protozoan infections in either species. Myxozoanosis was usually found in combination with other diseases and, except for 1 case, was restricted to weedy seadragons. Phaeohyphomycosis, nonmycobacterial bacterial infections, and trauma were also important but less frequent diagnoses. Intestinal coccidiosis was found in weedy but not leafy seadragons. Mineralization of the swim bladder was detected in 26 of 197 leafy seadragons and only 2 of 257 weedy seadragons. Although weedy and leafy seadragons share certain diseases of significance to exhibit populations, there are diseases unique to each species about which the veterinary pathologist, clinician, or diagnostician should be aware.

Induction of T-cell immunity against Ras oncoproteins by soluble protein or Ras-expressing Escherichia coli.

BACKGROUND: Point mutations in the ras proto-oncogene that activate its oncogenic potential occur in approximately 30% of human cancers. Previous studies have demonstrated that T-cell immunity against some forms of mutant Ras proteins could be elicited, and some effectiveness against tumors expressing activated Ras has been reported. PURPOSE: The goal of this study was to determine if immunization of mice with two forms of mutant Ras protein can induce high levels of Ras mutation-specific T-cell immunity in vitro and tumor regression in vivo. METHODS: Mice (BALB/c or C3H/HeJ) were immunized subcutaneously at 2-week intervals with purified Ras oncoproteins mixed with the immunologic adjuvants Antigen Formulation or QS-21, both of which have been shown to enhance the induction of T-cell-mediated immunity when included as components of soluble protein vaccines. In some experiments, mice were immunized directly with heat-killed Escherichia coli that had been induced to express one of the mutant Ras proteins. Spleen cells plus lymph node cells from Ras-immunized mice were tested in vitro for lysis of syngeneic Ras-expressing tumor cells and proliferation in response to mutant Ras peptides. For some of the cytolytic activity experiments, the spleen cells were grown under TH1 conditions (growth in presence of interleukin 2, interferon gamma, and an antibody directed against interleukin 4 to stimulate a cell-mediated immune response) or TH2 conditions (growth in presence of interleukins 2 and 4 to stimulate a humoral immune response). The specificity of immunity was examined in vivo by challenge of Ras-immunized mice with syngeneic tumor cells expressing mutant Ras oncoproteins (HaBalb, i.e., BALB/c mouse cells expressing Ras with arginine substituted at amino acid position 12 [Arg 12 Ras]; C3HL61, i.e., C3H/HeJ mouse cells expressing Ras with leucine substituted at position 61 [Leu 61 Ras]). Ten mice per group were used in each experiment. RESULTS: Proliferative and cytolytic T-cell responses directed against the Arg 12 Ras protein were generated in BALB/c mice, resulting in protection against challenge with cells expressing Arg 12 Ras and therapeutic benefit in mice bearing established tumors expressing this protein. In C3H/HeJ mice, high levels of cytolytic and proliferative responses were induced against Leu 61 Ras. Immunization with heat-killed E. coli genetically engineered to express Leu 61 Ras also led to the induction of anti-Ras T-cell immunity. T cells grown under TH1 conditions were cytolytic against Ras-transformed tumor cells, whereas those grown under TH2 conditions were not. CONCLUSIONS: Immunization as described here leads to Ras mutation-specific antitumor immunity in vitro and in vivo, with therapeutic efficacy in an established tumor model.

Cloning and analysis of MAGE-1-related genes.

The spectrum of MAGE gene expression in the human melanoma cell line DM150 was examined using reverse transcription polymerase chain reaction and cDNA cloning. We have isolated five full-length cDNAs from DM150 which were identified as MAGE-1, MAGE-3, MAGE-12 and two previously undescribed MAGE genes, MAGE-3b and MAGE-X2. DNA sequence analysis of the coding regions of the MAGE-3b and MAGE-X2 genes revealed 83% and 88% identity with MAGE-1, while MAGE-3b was 98% homologous with the full length MAGE-3 clone. The predicted amino acid sequences of MAGE-X2 and MAGE-3b contain consensus HLA-A1 peptide binding motifs, suggesting that, like MAGE-1, they may code for tumor-associated antigens. In addition, a nonamer peptide encoded by both the MAGE-3 and MAGE-12 genes was shown by direct binding studies to contain an aggretope for HLA-A2.

Evidence for in vivo Light-induced Synthesis of Ribulose-1,5-diP Carboxylase and Phosphoribulokinase in Greening Barley Leaves.

WHEN ACTINOMYCIN D, PUROMYCIN, STREPTOMYCIN, CHLORAMPHENICOL, AND CYCLOHEXIMIDE, KNOWN INHIBITORS OF PROTEIN SYNTHESIS, WERE APPLIED TO LEAVES OF INTACT SEEDLINGS OR DETACHED LEAVES OF BARLEY PRIOR TO THEIR GREENING, THE SAME GENERAL RESPONSE RESULTED: the light-induced increase in activity of ribulose 1,5-diphosphate carboxylase was prevented while that of phosphoribulokinase was only partially suppressed; synthesis of chlorophyll was arrested. This is taken as preliminary evidence that de novo synthesis of protein may be responsible for the observed increase in ribulose-1,5-diphosphate carboxylase activity during greening. However, other factors may be involved with the light-induced stimulation of phosphoribulokinase.Carbohydrate metabolites and substrates of the enzymes failed to induce the formation of ribulose-1,5-diphosphate carboxylase and phosphoribulokinase in the dark. No evidence was found for the presence of inhibitors in etiolated seedlings or activators in illuminated leaves of barley. Carboxylase activity almost equal to that of the illuminated water control was stimulated by MgCl(2) in the dark; MgCl(2) had no effect on the activity of the kinase.

Effects of Light Intensity on Photosynthetic Carboxylative Phase Enzymes and Chlorophyll Synthesis in Greening Leaves of Hordeum vulgare L.

The effects of various light intensities on in vivo increases in activities of phosphoriboisomerase, phosphoribulokinase and ribulose-1, 5-diP carboxylase and on synthesis of chlorophyll were studied in greening leaves of Hordeum vulgare L.Each enzyme was already present in dark-grown plants, but further increases in activities required both a light treatment of the intact plant and a favorable temperature. The amount of enzymatic activity and chlorophyll developed was governed by light intensity.Measured activities of phosphoriboisomerase and ribulose 1,5-diP carboxylase were highly correlated with synthesis of chlorophyll at all intensities studied. Measured activity of phosphoribulokinase was correlated with synthesis of chlorophyll only at saturating or near saturating light intensities. At decreasing light intensities the response curves of this enzyme differed from those of chlorophyll and of phosphoriboisomerase and ribulose-1, 5-diP carboxylase. A lag period of phosphoribulokinase increased with decreasing light intensity. After the lag period a rapid rate of increase occurred which did not level off during 48 hours of illumination. Thus, a different control mechanism may be operative in inducing increased activity of this enzyme.