The influence of three levels of vitamin E in the diet of pigs on the subcellular deposition of u-tocopherol in the muscle

HDGF (hepatoma-derived growth factor) and the HRPs (HDGF-related proteins) comprise a family of six proteins which display high identity in their N-terminus, but differ at the C-terminus. Here we investigate the patterns of expression of HDGF and HRP-3, by generating antisera specifically recognizing each growth factor. Whereas HRP-3 protein is expressed only in brain, HDGF can be found in a broad range of tissues, with highest levels in brain, testis, lung and spleen. The expression of HDGF and HRP-3 was found to be regulated during brain development, with highest levels around birth, followed by a decline until postnatal day 9. Interestingly, expression of HRP-3 increases again in adult brain. In situ hybridization and immunohistochemistry of cerebellar, cerebral and hippocampal brain slices showed that expression of both growth factors is not limited to areas of high proliferative activity. Both mRNAs and proteins are expressed in neuronal as well as glial cells. Immunocytochemistry of cultured neocortical neurons revealed that HDGF and HRP-3 can be found in the nucleus as well as the cytoplasm. HDGF is restricted to the neuronal soma, whereas HRP-3 can also be found in neurites. Thus the expression of HDGF and HRP-3 in differentiated cells, post-mitotic neurons and primary cultures of rat neocortex points to functions in brain that might not be limited to proliferation. In addition, their simultaneous expression in the same cell and their different subcellular localization in cultured neurons suggest different functions of HDGF and HRP-3 within single cells.

Twenty samples of rough rice (Oryza sativa) (unpolished kernels) collected during the 1995 harvest season from Arkansas (seven samples) and Texas (13 samples) were obtained from rice fields known to include plants with symptoms of Fusarium sheath rot putatively caused by Fusarium proliferatum. Samples were analyzed for fumonisin B1 (FB1) at three laboratories using three different extracting solvents by high-performance liquid chromatography (HPLC) or enzyme-linked immunosorbent assay (ELISA) methods. Forty percent of the samples were positive for FB1 at levels ≤4.3 μg/g by HPLC. The same samples contained FB1 at ≤3.6 μg/g when measured by an ELISA method. Most samples that were positive for FB1 were positive for fumonisin B2 (FB2) and fumonisin B3 (FB3) by HPLC at levels ≤1.2 μg/g. Very good agreement was obtained among the two laboratories using HPLC methods and the third using ELISA. Shelling of the unpolished rice results in hull and brown rice fractions. In a sample that contained 4.3 μg/g in whole kernels, the fumonisin level was very high in hulls (≤16.8 μg/g) and low in brown rice (≤0.9 μg/g). Milling of brown rice results in bran and white rice fractions. Fumonisins were found in bran at a level of ≤3.7 μg/g but were below the level of detection by HPLC in white rice. The presence of fumonisins (FB1, FB2, and FB3) was confirmed by fast atom bombardment/mass spectrometry. This is the first report of fumonisins in naturally contaminated rice in the United States.

Monoclonal antibodies were prepared against the fumonisins, a group of mycotoxins produced by Fusarium moniliforme. Splenic lymphocytes from mice immunized with a fumonisin B1-cholera toxin conjugate were fused with NS-1 myeloma cells, and six hybridomas were selected. A direct competitive ELISA was devised whereby fumonisin B1-peroxidase and free fumonisin B1 competed for antibody binding. The detection limit for fumonisin B1 in this assay was 50 ng/mL. Antibodies also cross-reacted with fumonisins BP and B3. Mean concentrations of fumonisins B1, BP, and B3 required to inhibit 50% antibody binding for the six clones were 630,1800, and 2300 ng/mL, respectively. When the antibodies were applied to the direct ELISA in spiked (5-25 pglg) feed, the average recovery was 103 % , with mean intra-and interassay coefficients of variation of 11 and 15%, respectively. These antibodies should find wide usage in the ELISA screening of fumonisins in foods, feeds, and tissues.

Direct fermentation of unhydrolyzed potato starch to ethanol by monocultures of an amylolytic fungus, Aspergilus niger, and cocultures ofA. niger and Saccharomyces cerevisiae was investigated. Amylolytic activity, rate and amount of starch utilization, and ethanol yields increased several-fold in coculture versus the monoculture due to the synergistic metabolic interactions between the species. Optimal ethanol yields were obtained in the pH range 5 to 6 and amylolytic activity was obtained in the pH range 5 to 8. Ethanol yields were maximal when fermentations were conducted anaerobically. Increasing S. cerevisiae inoculum in the coculture from 4 to 12% gave a dramatic increase in the rate of ethanol production, and ethanol yields of >96% of the theoretical maximum were obtained within 2 days of fermentation. These results indicate that simultaneous fermentation of starch to ethanol can be conducted efficiently by using cocultures of the amylolytic fungus A. niger and a nonamylolytic sugar fermenter, S. cerevisiae.