Cross-reactive anti-human monoclonal antibodies as a tool for B-cell identification in dogs and pigs

We have characterized a panel of commercially available anti-human monoclonal antibodies (mAbs) suitable for B-cell identification in pigs and dogs. The specificities of the mAbs were against CD20, CD21, CD22, and CD86. In addition to HM57, originally raised against human CD79alpha the broad cross-reactivity of which was documented more than 10 years ago, we recommend here a panel of several other mAbs as a useful tool for immunophenotyping and multicolor flow cytometry of canine and porcine B-lymphocytes. All six investigated antibodies did bind weakly to either canine or porcine lymphocytes (or both), but considerable weaker than for the human control cells. Four of them did bind to canine or porcine spleen section in immunohistochemistry. Monoclonal antibody against CD22 (clone RFB-4) was the only antibody in the tested panel the cross-reactivity of which was confirmed by Western blot. The advantages and limits of cross-reactive mAbs in studies on animal B-cells are discussed.     

松茸SCAR标记的建立

松茸群全球共报道15个种1变种,我国记载5个种1变种(臧穆,1990).长白山区有2种,即松茸(Tricholoma matsutake)和假松茸(T.bakamatsutake),同属于口蘑属.松茸是与松属(Pinus)树木共生的一种外生菌根菌,不仅菌肉肥厚、营养丰富、风味绝佳,而且具有抗癌作用(傅伟杰等,2005).     

The local immune response of mice after Helicobacter suis infection: strain differences and distinction with Helicobacter pylori

Helicobacter (H.) suis colonizes the stomach of pigs and is the most prevalent gastric non-H. pylori Helicobacter species in humans. Limited information is available on host immune responses after infection with this agent and it is unknown if variation in virulence exists between different H. suis strains. Therefore, BALB/c and C57BL/6 mice were used to compare colonization ability and gene expression of various inflammatory cytokines, as determined by real-time PCR, after experimental infection with 9 different H. suis strains. All strains were able to persist in the stomach of mice, but the number of colonizing bacteria at 59 days post inoculation was higher in stomachs of C57BL/6 mice compared to BALB/c mice. All H. suis strains caused an upregulation of interleukin (IL)-17, which was more pronounced in BALB/c mice. This upregulation was inversely correlated with the number of colonizing bacteria. Most strains also caused an upregulation of regulatory IL-10, positively correlating with colonization in BALB/c mice. Only in C57BL/6 mice, upregulation of IL-1β was observed. Increased levels of IFN-γ mRNA were never detected, whereas most H. suis strains caused an upregulation of the Th2 signature cytokine IL-4, mainly in BALB/c mice. In conclusion, the genetic background of the murine strain has a clear impact on the colonization ability of different H. suis strains and the immune response they evoke. A predominant Th17 response was observed, accompanied by a mild Th2 response, which is different from the Th17/Th1 response evoked by H. pylori infection.     

Entangling the spatial properties of laser beams

Position and momentum were the first pair of conjugate observables explicitly used to illustrate the intricacy of quantum mechanics. We have extended position and momentum entanglement to bright optical beams. Applications in optical metrology and interferometry require the continuous measurement of laser beams, with the accuracy fundamentally limited by the uncertainty principle. Techniques based on spatial entanglement of the beams could overcome this limit, and high-quality entanglement is required. We report a value of 0.51 for inseparability and 0.62 for the Einstein-Podolsky-Rosen criterion, both normalized to a classical limit of 1. These results are a conclusive optical demonstration of macroscopic position and momentum quantum entanglement and also confirm that the resources for spatial multimode protocols are available.     

An oncogene-induced DNA damage model for cancer development

Of all types of DNA damage, DNA double-strand breaks (DSBs) pose the greatest challenge to cells. One might have, therefore, anticipated that a sizable number of DNA DSBs would be incompatible with cell proliferation. Yet recent experimental findings suggest that, in both precancerous lesions and cancers, activated oncogenes induce stalling and collapse of DNA replication forks, which in turn leads to formation of DNA DSBs. This continuous formation of DNA DSBs may contribute to the genomic instability that characterizes the vast majority of human cancers. In addition, in precancerous lesions, these DNA DSBs activate p53, which, by inducing apoptosis or senescence, raises a barrier to tumor progression. Breach of this barrier by various mechanisms, most notably by p53 mutations, that impair the DNA damage response pathway allows cancers to develop. Thus, oncogene-induced DNA damage may explain two key features of cancer: genomic instability and the high frequency of p53 mutations.     

Summary of workshop findings for porcine B-cell markers.

Based on cluster groups from the first-round analyses of the Third International Swine CD Workshop, 38 monoclonal antibodies (MAbs) including eight internal controls were analysed by flow cytometry (FCM) and immunohistochemistry (IH) in the second-round analysis of the B-cell section of this workshop. Targets in this section included peripheral blood lymphocytes and cells isolated from ileal Peyer's patches (PP), mesenteric lymph nodes (MLN) of adult animals, bone marrow cells from newborn piglets and thymus cells isolated from foetuses at day 105 of gestation.Immunohistochemistry of these 38 MAbs identified four sets, whose ligands were co-expressed with CD21, which showed a tissue distribution compatible with specificity for cells including those of the B-cell lineage. Another group of miscellaneous antibodies appeared to identify other cells, several antibodies were negative. Two-colour flow cytometry (2C-FCM) was carried out by pairing each antibody of interest with antibodies to SWC7, CD21, sIgM and a polyclonal rabbit anti-swine immunoglobulin antiserum (RaSwIg).The anti-CD21 MAb BB6-11C9 (no. 20) and IAH-CC51 (no. 19), established in previous workshops, as well as the cross-reactive anti-human CD21 B-1y4 (no. 146), clustered together in FCM analyses of the first round and showed similar cellular distribution in IH. A further cluster was formed by the standard CC55 (no. 55) and 2A10/8 (no. 102) submitted as SWC7 specific. The second SWC7 standard 2F6/8 (no. 100) clustered separately, but IH showed an identical pattern of reactivity to the other SWC7 MAb.Unfortunately, this work could not identify any other novel clusters with specificity for B-cells, as the statistical clustering of other MAbs could not be substantiated by IH or subsequent two-colour-FCM work. However, we could identify MAb with similar cellular distribution. The ligands for the cross-reactive anti-human CD40 G28.5 (no. 25) and STH224 (no. 153) were expressed on very similar targets, similarly the ligands for the MAb pair JM1H1 (no. 139) with BB6-10A10 (no. 142) and the MAb pair 3F7/11 (no. 115) with 1C2F10 (no. 187).     

Laboratory Study of Stormwater Biofiltration in Low Temperatures: Total and Dissolved Metal Removals and Fates

Stormwater biofilters, which are recommended for application in both Water-Sensitive Urban Design and Low Impact Development, can remove up to 80% or 90% of total metals found in stormwater. However, their winter operation is a common concern. That was addressed in this study by investigating the metal removal effectiveness of replicate laboratory biofilter mesocosms at 2°C, 8°C and 20°C. As recommended for cold climate bioretention, coarse filter media were implemented and in the top 100 mm layer topsoil was added to increase the sorption capacity. Cd, Cu, Pb and Zn concentrations measured in the biofilter effluent were far below those in the influent and this significantly improved the treated stormwater quality. Contrary to a common notion that coarse media in the main filter body impair dissolved metal sorption, satisfactory removals of dissolved metals were found in this study with most metal burdens retained in the top layer of the filter in which the sorption capacity was enhanced by topsoil. Some metal uptake by the plants was also detected. Temperature did not affect Cd, Pb and Zn removals in general, but Cu removals increased with decreasing temperatures. This was explained by increased biological activities in the filters at warmer temperatures, which may have led to an increased release of Cu with dissolved organic matter originating from root turnover and decomposition of organic litter and debris. Furthermore, plant uptake and biofilm adsorption may also be influenced by temperature. However, even in the worst case (i.e. at 20°C), Cu was removed effectively from the stormwater. Further research needs were identified including the effects of road salts on stormwater biofiltration during the winter period.     

Regulation of replication fork progression through histone supply and demand

DNA replication in eukaryotes requires nucleosome disruption ahead of the replication fork and reassembly behind. An unresolved issue concerns how histone dynamics are coordinated with fork progression to maintain chromosomal stability. Here, we characterize a complex in which the human histone chaperone Asf1 and MCM2-7, the putative replicative helicase, are connected through a histone H3-H4 bridge. Depletion of Asf1 by RNA interference impedes DNA unwinding at replication sites, and similar defects arise from overproduction of new histone H3-H4 that compromises Asf1 function. These data link Asf1 chaperone function, histone supply, and replicative unwinding of DNA in chromatin. We propose that Asf1, as a histone acceptor and donor, handles parental and new histones at the replication fork via an Asf1-(H3-H4)-MCM2-7 intermediate and thus provides a means to fine-tune replication fork progression and histone supply and demand.     

Effect of combined nitrogen and sulphur fertilization on yield and qualitative parameters of Camelina sativa [L.] Crtz. (false flax)

Abstract

Camelina (Camelina sativa (L.) Crtz., false flax) is described as a species requiring fewer inputs than other oilseed crops thus making it an interesting alternative in sustainable cropping systems. As information on the combined effects of nitrogen and sulphur on camelina yield and quality parameters is meagre, a pot fertilization experiment was carried out with nitrogen applied as NH₄NO₃ at three increasing rates equivalent to a range from 63 up to 127 kg ha^?1. These treatments were combined with sulphur additions applied as (NH₄)₂SO₄ to achieve a soil sulphate content of 25 or 45 mg kg^?1, respectively (equivalent to 75 and 135 kg S ha^?1). The medium and high nitrogen rates combined with the low sulphur fertilization level increased the number of branches per plant compared with the lowest nitrogen fertilization at the same sulphur rate. Camelina seed yield increased with increasing nitrogen doses at the same low sulphur level, whereas straw yield increased only at the highest rate of nitrogen. Thousand-seed mass increased at the highest nitrogen dose and with the low sulphur application rate. In addition, the increase in nitrogen fertilization reduced seed oil content from 39.8% to 37.1%. A highly negative correlation was observed between oil and protein content of seeds. Nitrogen fertilization increased total oil yield and total protein yield. Differences in crop parameters between the sulphur treatments were not significant statistically, although the higher sulphur treatment tended to increase seed yield as well as oil and protein content compared with the low sulphur treatment. Thus, the combined application of N and S as mineral fertilizers is only recommended when growing camelina on sulphur-deficient soils if the aim is to achieve both high oil and protein production.