Application of Protein Feed Processed by Microbial Fermentation to Dairy Cow

Methionine(Met)and lysine(Lys)have been reported as the first two limiting amino acids(AA)for maximum milk yield and milk protein production.Supplying these AA may improve microbial protein synthesis and therefore improve milk production without adding excess N to the environment.This observation utilized fermented soybean meal(SBM),cottonseed meal(CSM),rapeseed meal(RSM)and corn by Bacillus subtilis 168 and Leuconostoc mesenteroides as core feedstuffs to produce special biological protein feed for dairy cow.The results showed that the milk production,milk protein percentage,milk fat percentage and milk DM percentage of test groups in trial period were significantly more than those of the control group(P<0.01),the results showed that adding fermenting protein feed in dairy cow diets could significantly improve milk yield,milk protein and milk fat content.The economic benefits of actual application were analyzed,the group of 0.5%was the best compared with the other groups.     

Immune responses and enhanced disease resistance in Chinese drum, Miichthys miiuy (Basilewsky), after oral administration of live or dead cells of Clostridium butyrium CB2

Clostridium butyrium CB2 isolated from chickens was tested as a potential fish probiotic in the Chinese drum, Miichthys miiuy . Fish were fed live (CB), dead CB2 (D-CB) cells (10⁸ cells g⁻¹) or spent culture supernatant (SCS), for 30 days and challenged with Vibrio anguillarum or Aeromonas hydrophila . Survival was higher in both the CB and the CB-D fed groups, but the SCS group was not significantly different from the control. After feeding live or dead CB2 cells, there was increase in phagocytic activity of the head kidney macrophages, the lysozyme activity of serum and gut mucosa and immunoglobulin (Ig) level. The SCS group showed no obvious change in immune parameters. The results suggest that live or dead Clostridium butyrium CB2 has an immunomodulatory effect on fish.     

The immune response and protective efficacy of oral alginate microparticle Aeromonas sobria vaccine in soft-shelled turtles (Trionyx sinensis)

Aeromonas sobria is an important bacterial pathogen of soft-shelled turtles (Trionyx sinensis). The effect of an oral alginate microparticle A. sobria vaccine (MAV) on immune response and protection efficacy of soft-shelled turtles was investigated. The formalin-killed cultures of A. sobria strain Z-1 (FKC) were incorporated into alginate microparticles, and the encapsulated vaccine was orally used for immunizing turtles. The serum agglutinating antibody titers, bactericidal activity of blood leucocytes and relative percent survival (RPS) post-challenge with virulent live bacteria of control and immunized turtles were detected. The serum agglutinating antibody titers and bactericidal activity of the leucocytes in the turtles orally immunized with MAV reached a level equivalent to those of the turtles injected with FKC and were significantly higher than those of control group. The RPS of turtles immunized orally with the MAV and intraperitoneally with FKC was 89.5 and 94.7%, respectively, while 95% of the control turtles had died. These results demonstrated that oral alginate microparticle A. sobria vaccine was immunogenic, and could confer protective immunity in soft-shelled turtles.     

New alleles of chicken CD8 alpha and CD3d found in Chinese native and western breeds

Chinese native chicken breeds provide useful resources for the study of genetic diversity. In this study, the alleles of CD8 alpha and CD3d cDNA from Chinese native and introduced western breeds of chicken were analyzed at the sequence level. Six alleles were found, due to 13 amino acid replacements in the extracellular domain of the CD8 alpha sequence. There were four alleles detected in the Chinese strains, and alleles 5 and 6 were identified for the first time. Allele 6 was shared by Langshan, Beijing Fatty and Recessive White Feather chickens. Allele 2, found in the Bigbone strain, was the same as that found in the Leghorn strain H.B15.H7, and allele 3 in the Xianju breed was also the same as in the Leghorn strain H.B15.H12. Two Leghorn lines (RPL line 7 and AY519197) and the Camellia possessed an allele (alleles 1, 4 and 5), respectively, that was not found in the other lines. Nine out of 13 amino acid replacements were situated in the putative major histocompatibility complex (MHC) class I binding CDR1 (positions 30, 33 and 34), CDR2 (positions 58, 62, 63 and 65) and CDR3 (positions 90 and 106). Except for the Xianju breed, the CD8 alpha cDNA of Chinese native chicken breeds shared high homology. Two alleles were found in CD3d. Three additional nucleotides were found at positions 64, 65 and 66 in the newly discovered allele 2. This led to a difference of four amino acids (at residues 22, 23, 24 and 25) in the extracellular domain of CD3d cDNA from the Gushi, Recessive White Feather and ISA chickens compared with these of the White Leghorn and T11.15 (NM_205512). Five hybridoma clones (1C9, 1H5, 4B11, 6G5 and 13C5) against chicken CD8 alpha were generated by DNA immunization. Two monoclonal antibodies (mAbs), 6G5 and 4B11, showed reactivity to the splenocytes from five Chinese native chicken breeds, the Recessive White Feather chicken and the Leghorn (AY519197), while mAbs 1C9, 1H5 and 13C5 showed no reaction with these breeds.     

Detection of genetically modified maize MON810 and NK603 by multiplex and real-time polymerase chain reaction methods

In this study, the event-specific primers for insecticide-resistant maize, MON810, and herbicide-tolerance maize, NK603, have been designed. Simplex PCR and multiplex PCR detection method have been developed. The detection limit of the multiplex PCR is 0.5% for MON810 and NK603 in 50 ng of the template for one reaction. Quantitative methods based on real-time quantitative PCR were developed for MON810 and NK603. Plasmid pMulM2 as reference molecules for the detection of MON810 and NK603 was constructed. Quantification range was from 0.5 to 100% in 100 ng of the DNA template for one reaction. The precision of real-time Q-PCR detection methods, expressed as coefficient of variation for MON810 and NK603 varied from 1.97 to 8.01% and from 3.45 to 10.94%, respectively. The range agreed with European interlaboratories test results (25%). According to the results, the methods for quantitative detection of genetically modified maize were acceptable.     

Induction of apoptosis by the oolong tea polyphenol theasinensin A through cytochrome c release and activation of caspase-9 and caspase-3 in human U937 cells

This study examined the growth inhibitory effects of theasinensin A (from oolong tea) and black tea polyphenols, including theaflavin (TF-1), a mixture (TF-2) of theaflavin-3-gallate (TF-2a) and theaflavin-3'-gallate (TF-2b), and theaflavin-3,3'-digallate (TF-3) in human cancer cells. Theasinensin A, TF-1, and TF-2 displayed strong growth inhibitory effects against human histolytic lymphoma U937, with estimated IC50 values of 12 microM, but were less effective against human acute T cell leukemia Jurkat, whereas TF-3 and (-)-epigallocatechin-3-gallate (EGCG) had lower activities. The molecular mechanisms of tea polyphenol-induced apoptosis as determined by annexin V apoptosis assay, DNA fragmentation, and caspase activation were further investigated. Loss of membrane potential and reactive oxygen species (ROS) generation were also detected by flow cytometry. Treatment with tea polyphenols caused rapid induction of caspase-3, but not caspase-1, activity and stimulated proteolytic cleavage of poly(ADP-ribose) polymerase (PARP). Pretreatment with a potent caspase-3 inhibitor, Z-Asp-Glu-Val-Asp-fluoromethyl ketone, inhibited theasinensin A induced DNA fragmentation. Furthermore, it was found that theasinensin A induced loss of mitochondrial transmembrane potential, elevation of ROS production, release of mitochondrial cytochrome c into the cytosol, and subsequent induction of caspase-9 activity. These results indicate that theasinensin A allows caspase-activated deoxyribonuclease to enter the nucleus and degrade chromosomal DNA and induces DFF-45 (DNA fragmentation factor) degradation. The results suggest that induction of apoptosis by theasinensin A may provide a pivotal mechanism for their cancer chemopreventive function.     

Basis for structural diversity in homologous RNAs

Large RNA molecules, such as ribozymes, fold with well-defined tertiary structures that are important for their activity. There are many instances of ribozymes with identical function but differences in their secondary structures, suggesting alternative tertiary folds. Here, we report a crystal structure of the 161-nucleotide specificity domain of an A-type ribonuclease P that differs in secondary and tertiary structure from the specificity domain of a B-type molecule. Despite the differences, the cores of the domains have similar three-dimensional structure. Remarkably, the similar geometry of the cores is stabilized by a different set of interactions involving distinct auxiliary elements.     

Giant piezoelectricity on Si for hyperactive MEMS

Microelectromechanical systems (MEMS) incorporating active piezoelectric layers offer integrated actuation, sensing, and transduction. The broad implementation of such active MEMS has long been constrained by the inability to integrate materials with giant piezoelectric response, such as Pb(Mg(1/3)Nb(2/3))O(3)-PbTiO(3) (PMN-PT). We synthesized high-quality PMN-PT epitaxial thin films on vicinal (001) Si wafers with the use of an epitaxial (001) SrTiO(3) template layer with superior piezoelectric coefficients (e(31,f) = -27 ± 3 coulombs per square meter) and figures of merit for piezoelectric energy-harvesting systems. We have incorporated these heterostructures into microcantilevers that are actuated with extremely low drive voltage due to thin-film piezoelectric properties that rival bulk PMN-PT single crystals. These epitaxial heterostructures exhibit very large electromechanical coupling for ultrasound medical imaging, microfluidic control, mechanical sensing, and energy harvesting.     

Polycrystalline CVD Diamond Films with High Electrical Mobility

Advances in the deposition process have led to dramatic improvements in the electronic properties of polycrystalline diamond films produced by chemical vapor deposition (CVD). It is now possible to produce CVD diamond with properties approaching those of IIa natural diamonds. The combined electron-hole mobility, as measured by transient photoconductivity at low carrier density, is 4000 square centimeters per volt per second at an electric field of 200 volts per centimeter and is comparable to that of the best single-crystal IIa natural diamonds. Carrier lifetimes measured under the same conditions are 150 picoseconds for the CVD diamond and 300 picoseconds for single-crystal diamond. The collection distance at a field of 10 kilovolts per centimeter is 15 micrometers for the CVD diamond as compared to 30 micrometers for natural diamonds. The electrical qualities appear to correlate with the width of the diamond Raman peak. Also, although the collection distance at the highest fields in the films nearly equals the average grain size, there is no evidence of deleterious grain boundary effects.