Transmembrane TNF is sufficient to initiate cell migration and granuloma formation and provide acute, but not long-term, control of Mycobacterium tuberculosis infection

TNF is critical for immunity against Mycobacterium tuberculosis infection; however, the relative contributions of the soluble and transmembrane forms of TNF in this immunity are unknown. Using memTNF mice, which express only the transmembrane form of TNF, we have addressed this question. Wild-type (WT), TNF-/-, and transmembrane TNF (memTNF) mice were infected with M. tuberculosis by aerosol. TNF-/- mice developed overwhelming infection with extensive pulmonary necrosis and died after only 33 days. memTNF mice, like WT mice, contained bacterial growth for over 16 wk, developed an Ag-specific T cell response, and initially displayed compact granulomas, comprised of both lymphocytes and macrophages. Expression of mRNA for the chemokines CXCL10, CCL3, CCL5, and CCL7 was comparable in both WT and memTNF mice. As the infection progressed, however, the pulmonary lesions in memTNF mice became larger and more diffuse, with increased neutrophil accumulation and necrosis. This was accompanied by increased influx of activated memory T cells into the lungs of memTNF mice. Eventually, these mice succumbed to infection with a mean time to death of 170 days. The expression of memTNF on T cells is functionally important because the transfer of T cells from memTNF, but not TNF-/- mice, into either RAG-/- or TNF-/- mice conferred the same survival advantage on the M. tuberculosis-infected recipient mice, as the transfer of WT T cells. Therefore, memTNF, in the absence of soluble TNF, is sufficient to control acute, but not chronic, M. tuberculosis infection, in part through its expression on T cells.     

中国斑翅跳小蜂属一新种(膜翅目,跳小蜂科)

记述了育自上海桃树上1种盾蚧中的斑翅跳小蜂属1新种:上海斑翅跳小蜂Epitetracnemus shanghaiensis sp.nov..文中给出了新种的形态特征图和该属分种检索表.研究标本保存于中国科学院上海生命科学研究院植物生理生态研究所.     

优化萝卜基因组DNA RAPD-PCR反应体系的正交设计法

以CTAB微量提取方法提取10个红萝卜雄性不育系A单株的基因组DNA,等浓度混合成基因池。以其为模板,用正交设计方法论定RAPD-PCR反应体系的各影响因子,用随机引物S42（5’GGACCCAACC3’）优化萝卜雄性不育系A基因组DNARAPD,PCR反应体系,16次试验即可获得最佳的RAPD-PCR反应体系。     

Geographical variation in species diversity: A comparison of marine polychaetes and nematodes

The aim of this study was to determine if marine species diversity was influenced by geographical location and whether it was higher at lower latitudes. Artificial collectors (made of nylon pan scourers) were employed as a standard substratum for the colonisation of marine invertebrates inhabiting subtidal (12 to 15 m) hard, rocky bottom substrata. These artificial substrate units (ASUs) were deployed at different latitudes including northern and southern temperate (South West England, UK and New Zealand), tropical (Trinidad and Tobago, West Indies) and polar (Signy Island, Antarctica) areas. The polychaetes, representative of the macrofauna and the nematodes, representative of the meiofauna fractions of the total invertebrate fauna collected were analysed.Neither polychaete nor nematode species diversity showed a trend based on latitude and each taxon showed a different pattern of diversity variation in relation to location. Polychaete diversity varied from area to area with highest species diversity occurring in the southern temperate (New Zealand). Nematode species diversity however was similar for the northern and southern temperate (UK and New Zealand) and the tropical area (Trinidad and Tobago). Thus, although the number of locations studied was limited, these data do not conform to a gradient in species diversity with latitude as has been previously supposed. The success of ASUs to compare species diversities in standardised habitat units augurs well for their future use in other ecological areas such as biogeographical or pollution studies.     

Exploring interactions by second-stage community analyses

Many biological data sets, from field observations and manipulative experiments, involve crossed factor designs, analysed in a univariate context by higher-way analyses of variance which partition out ‘main’ and ‘interaction’ effects. Indeed, tests for significance of interactions among factors, such as differing Before-After responses at Control and Impact sites, are the basis of the widely used BACI strategy for detecting impacts in the environment. There are difficulties, however, in generalising simple univariate definitions of interaction, from classic linear models, to the robust, non-parametric multivariate methods that are commonly required in handling assemblage data. The size of an interaction term, and even its existence at all, depends crucially on the measurement scale, so it is fundamentally a parametric construct. Despite this, certain forms of interaction can be examined using non-parametric methods, namely those evidenced by changing assemblage patterns over many time periods, for replicate sites from different experimental conditions (types of ‘Beyond BACI’ design) - or changing multivariate structure over space, at many observed times. Second-stage MDS, which can be thought of as an MDS plot of the pairwise similarities between MDS plots (e.g. of assemblage time trajectories), can be used to illustrate such interactions, and they can be formally tested by second-stage ANOSIM permutation tests. Similarities between (first-stage) multivariate patterns are assessed by rank-based matrix correlations, preserving the fully non-parametric approach common in marine community studies. The method is exemplified using time-series data on corals from Thailand, macrobenthos from Tees Bay, UK, and macroalgae from a complex recolonisation experiment carried out in the Ligurian Sea, Italy. The latter data set is also used to demonstrate how the analysis copes straightforwardly with certain repeated-measures designs.     

Heritable transgenesis of Parastrongyloides trichosuri: a nematode parasite of mammals

Germline transformation of a parasitic nematode of mammals has proven to be an elusive goal. We report here the heritable germline transformation of Parastrongyloides trichosuri, a nematode parasite whose natural hosts are Australian possums of the genus Trichosurus. This parasite can undergo multiple free-living life cycles and these replicative cycles can be maintained indefinitely in the laboratory. Transformation was achieved by microinjection of DNA into the ovary syncytium of either free-living or parasitic adult females. By selecting for the transgenic progeny of successive free-living life cycles, it was possible to establish and maintain transgenic lines. All three transgenic lines tested were shown capable of establishing patent infections in possums and to transmit the functional transgene to their progeny. The transgene, driven by the Pt hsp-1 promoter, was constitutively expressed in intestinal cells at all stages of both parasitic and free-living life cycles, although gene silencing appears to occur in some transgenic progeny. This is the first report of heritable transgenesis in a parasitic nematode of a mammal and we discuss a variety of previously inaccessible experimental avenues that will now be possible with this powerful model system.     

强光照和全黑暗条件下荒漠沙蜥视网膜内GAP-43表达的免疫组织化学

采用免疫组织化学技术研究了在强光照和全黑暗条件下荒漠沙蜥(Phrynocephalus prezewalskic)视网膜内生长相关蛋白GAP-43的表达变化。结果表明,在正常光照条件下,视网膜内GAP-43阳性表达部位主要存在于内网层;强光照条件下,GAP-43免疫染色部位主要出现在内网层、节细胞层和内核层的部分细胞核。在全黑暗条件下,在视纤维层和内网层呈阳性染色;提示视网膜在不同环境条件下GAP-43的不同定位,可能与其在相应的环境下参与不同的视觉功能有关。     

No evidence from FTIR difference spectroscopy that glutamate-189 of the D1 polypeptide ligates a Mn ion that undergoes oxidation during the S0 to S1, S1 to S2, or S2 to S3 transitions in photosystem II

In the recent X-ray crystallographic structural models of photosystem II, Glu189 of the D1 polypeptide is assigned as a ligand of the oxygen-evolving Mn(4) cluster. To determine if D1-Glu189 ligates a Mn ion that undergoes oxidation during one or more of the S(0) --> S(1), S(1) --> S(2), and S(2) --> S(3) transitions, the FTIR difference spectra of the individual S-state transitions in D1-E189Q and D1-E189R mutant PSII particles from the cyanobacterium Synechocystis sp. PCC 6803 were compared with those in wild-type PSII particles. Remarkably, the data show that neither mutation significantly alters the mid-frequency regions (1800-1200 cm(-)(1)) of any of the FTIR difference spectra. Importantly, neither mutation eliminates any specific symmetric or asymmetric carboxylate stretching mode that might have been assigned to D1-Glu189. The small spectral alterations that are observed are similar in amplitude to those that are observed in wild-type PSII particles that have been exchanged into FTIR analysis buffer by different methods or those that are observed in D2-H189Q mutant PSII particles (the residue D2-His189 is located >25 A from the Mn(4) cluster and accepts a hydrogen bond from Tyr Y(D)). The absence of significant mutation-induced spectral alterations in the D1-Glu189 mutants shows that the oxidation of the Mn(4) cluster does not alter the frequencies of the carboxylate stretching modes of D1-Glu189 during the S(0) --> S(1), S(1) --> S(2), or S(2) --> S(3) transitions. One explanation of these data is that D1-Glu189 ligates a Mn ion that does not increase its charge or oxidation state during any of these S-state transitions. However, because the same conclusion was reached previously for D1-Asp170, and because the recent X-ray crystallographic structural models assign D1-Asp170 and D1-Glu189 as ligating different Mn ions, this explanation requires that (1) the extra positive charge that develops on the Mn(4) cluster during the S(1) --> S(2) transition be localized on the Mn ion that is ligated by the alpha-COO(-) group of D1-Ala344 and (2) any increase in positive charge that develops on the Mn(4) cluster during the S(0) --> S(1) and S(2) --> S(3) transitions be localized on the one Mn ion that is not ligated by D1-Asp170, D1-Glu189, or D1-Ala344. An alternative explanation of the FTIR data is that D1-Glu189 does not ligate the Mn(4) cluster. This conclusion would be consistent with earlier spectroscopic analyses of D1-Glu189 mutants, but would require that the proximity of D1-Glu189 to manganese in the X-ray crystallographic structural models be an artifact of the radiation-induced reduction of the Mn(4) cluster that occurred during the collection of the X-ray diffraction data.     

A pharmacological study of NLP-12 neuropeptide signaling in free-living and parasitic nematodes

NLP-12a and b have been identified as cholecystokinin/sulfakinin-like neuropeptides in the free-living nematode Caenorhabditis elegans. They are suggested to play an important role in the regulation of digestive enzyme secretion and fat storage. This study reports on the identification and characterization of an NLP-12-like peptide precursor gene in the rat parasitic nematode Strongyloides ratti. The S. ratti NLP-12 peptides are able to activate both C. elegans CKR-2 receptor isoforms in a dose-dependent way with affinities in the same nanomolar range as the native C. elegans NLP-12 peptides. The C-terminal RPLQFamide sequence motif of the NLP-12 peptides is perfectly conserved between free-living and parasitic nematodes. Based on systemic amino acid replacements the Arg-, Leu- and Phe- residues appear to be critical for high-affinity receptor binding. Finally, a SAR analysis revealed the essential pharmacophore in C. elegans NLP-12b to be the pentapeptide RPLQFamide.