Functional secretion of a type 1 antifreeze protein analogue by optimization of promoter, signal peptide, prosequence, and terminator in Lactococcus lactis

Lactococcus lactis is a food-grade microorganism of major commercial importance. Antifreeze protein is a potent cryogenic protection agent for the cryopreservation of food and pharmaceutical materials. In this study, extracellular expression of a novel recombinant type I antifreeze protein analogue (rAFP) in L. lactis was optimized. An efficient SlpA promoter (P SlpA) was fused to various signal peptides (SPs) and propeptide sequences to examine the extracellular expression levels of rAFP. An efficient signal peptide, SP sacB, fused to prosequence AE, enabled higher extracellular rAFP production; use of the SlpA terminator (Ter SlpA) was a further improvement. The extracellularly expressed rAFP successfully inhibited ice recrystallization and is thus potentially applicable for cryogenic preservation.     

Health of non-ambulatory, non-injured pigs at processing

Loss of pigs during or after transport is a welfare concern, but also an economic concern for producers. Transport losses include animals that are dead on arrival (DOA) at the plant, pigs that are injured, and pigs which are not obviously injured but unwilling or unable to walk (non-ambulatory, non-injured or NANI). The objective of this research was to assess the health of non-ambulatory, non-injured (NANI) pigs relative to control pigs at the processing plant by looking at a range of measures, including complete blood chemistry, anatomy, and pathology to determine potential factors associated with pigs going down. Data were collected from NANI and control pigs at five plants in the midwest USA. Feet and legs and internal organs were inspected and the severity of the pathology scored. Alveolar macrophages were collected and counted. Blood was collected for analysis of hematology, blood chemistry and cortisol concentrations. Titers to common porcine respiratory viruses were measured in pigs from one plant. Hoof and pad problems did not differ overall between control and NANI pigs, however the percentage of severe foot problems was greater (P < 0.05) in NANI compared with control pigs at plants A and E. The percentage of total ulcers, rhinitis, and empty stomachs differed (P < 0.05) between control and NANI pigs at individual plants, but not overall. Blood hematology and chemistry differed (P < 0.05) between NANI and control pigs. Cortisol concentrations did not differ between NANI and control pigs. Titers to swine influenza virus (SIV) H1N1 and H3N2 and porcine circovirus (PCV) were lower (P < 0.01) among NANI compared with control pigs. However, more (P < 0.01) NANI pigs were positive for SIV H1N1 and H3N2 compared with control pigs. Blood hematology, chemistry, and pathology indicate a large difference between NANI and controls pigs. No single health problem was higher among NANI pigs compared to plant-matched control pigs. Rather, several problems appear to contribute to pigs becoming NANI which may differ from one plant too another.     

The immune response of sheep to larval challenge with Ostertagia circumcincta and O. ostertagi.

One of three groups of sheep was challenged twice-weekly with infective-stage larvae (L3) of the sheep parasite O. circumcincta, another with the cattle parasite O. ostertagi while the third received no larval challenge. Positive faecal egg counts (FEC) and a rise in plasma pepsinogen levels were observed only in those animals given O. circumcincta. Anti-O. circumcincta L3 IgG titres were rapidly elevated during parasite challenge with either O. circumcincta or O. ostertagi. Throughout the experiment, no rise in anti-adult IgG titres or eosinophil numbers was observed in peripheral blood in any group. On evidence of self-cure of the trickle-infection, determined by a reduction in FEC, all groups were drenched and challenged with 15,000 O. circumcincta L3. No effect of previous challenge on parasite establishment or FEC was observed, although egg viability was significantly reduced in both groups given prior challenge. Significant differences in adult female worm length were observed between groups. Those recovered from animals previously challenged with O. circumcincta were shorter than from those given O. ostertagi which were in turn shorter than those from previously unchallenged animals. In utero egg counts were significantly lower in worms from animals previously challenged with O. circumcincta than in those from unchallenged control animals. The results indicate that a level of immunity to O. circumcincta can be conferred by exposure to O. ostertagi.     

The myrobalan (<Emphasis Type="Italic">Prunus cerasifera</Emphasis> L.): a useful diploid model for studying the molecular genetics of self-incompatibility in plums

A series of PCR methods were used to detect S-RNase alleles and SFB alleles and to determine S-genotypes in 25 accessions of myrobalan (Prunus cerasifera L.). Firstly, primers flanking the polymorphic second intron were used to identify S-RNases in agarose gels. These primers amplified one or two bands per accession in 25 accessions. Then consensus primers were designed for amplifying the polymorphic first intron, unique to Prunus S-RNases, for automated fluorescent detection. Each accession produced one or two peaks. New primers were then developed to amplify the intron in the SFB gene, for detection by fluorescence. Cross-referencing PCR bands and peaks indicated 15 S-alleles were present in the 25 accessions. Cloning, sequencing and comparison with published data indicated that the amplified products were S-RNase alleles. Sequence information was used to design primers specific for each S-RNase. Full and consistent S-genotypes were obtained by cross-comparing PCR data for 23 of the 25 accessions, and two accessions appeared to have a single allele. Pollen-tube microscopy indicated function of some but not all of the S-alleles sequenced.     

Myoblast cell interaction with polydopamine coated liposomes

Liposomes are widely used, from biosensing to drug delivery. Their coating with polymers for stability and functionalization purposes further broadens their set of relevant properties. Poly(dopamine) (PDA), a eumelanin-like material deposited via the "self"-oxidative polymerization of dopamine at mildly basic pH, has attracted considerable interest in the past few years due to its simplicity, flexibility yet fascinating properties. Herein, we characterize the coating of different types of liposomes with PDA depending on the presence of oleoyldopamine in the lipid bilayer and the dopamine hydrochloride concentration. Further, the interaction of these coated liposomes in comparison to their uncoated counterparts with myoblast cells is assessed. Their uptake/association efficiency with these cells is determined. Further, their dose-dependent cytotoxicity with and without entrapped hydrophobic cargo (thiocoraline) is characterized. Taken together, the reported results demonstrate the potential of PDA coated liposomes as a tool in biomedical applications.     

中国大豆花叶病抗源和抗性鉴别寄主的鉴定与评价

收集大豆抗大豆花叶病(SMV)抗源和鉴别寄主40份,通过接种Cho和Goodman划分的抗大豆花叶病毒株系SMV G1-G7,了解这些材料对该株系的抗性反应.同时比较了其中部分材料对中国学者划分的SMV Sc1-Sc17株系的抗性反应.结果感病材料无论对SMV G1-G7株系,还是SMV Sc1-Sc17株系均表现感病;但是,齐黄1、科丰1、早18和8101等对SMV G1-G7株系均表现抗病的材料,对SMV Sc1-Sc17株系却表现出部分抗病;而另外一些材料,如:诱变30、徐豆1、文丰5、铁6915、齐黄10和Harosoy等对SMV G1-G7株系的抗性反应却与北美的鉴别寄主相同.结果表明:无论是对 SMV G1-G7株系,还是对SMV Sc1-Sc17株系,抗病材料的抗性遗传基础是相似的;中国一些大豆花叶病毒株系的致病力强于国外的株系.因此,结合国外的SMV株系鉴定系统,创建一套统一的SMV株系鉴定系统是可行的.