Clinical outcome following acute ischaemic stroke relates to both activation and autoregulatory inhibition of cytokine production

Background

As critical mediators of local and systemic inflammatory responses, cytokines are produced in the brain following ischaemic stroke. Some have been detected in the circulation of stroke patients, but their role and source is unclear. Focusing primarily on interleukin(IL)-1-related mechanisms, we serially measured plasma inflammatory markers, and the production of cytokines by whole blood, from 36 patients recruited within 12 h and followed up to 1 year after acute ischaemic stroke (AIS).

Results

Admission plasma IL-1 receptor antagonist (IL-1ra) concentration was elevated, relative to age-, sex-, and atherosclerosis-matched controls. IL-1β, soluble IL-1 receptor type II, tumour necrosis factor (TNF)-α, TNF-RII, IL-10 and leptin concentrations did not significantly differ from controls, but peak soluble TNF receptor type I (sTNF-RI) in the first week correlated strongly with computed tomography infarct volume at 5–7 days, mRS and BI at 3 and 12 months. Neopterin was raised in patients at 5–7 d, relative to controls, and in subjects with significant atherosclerosis. Spontaneous IL-1β, TNF-α and IL-6 gene and protein expression by blood cells was minimal, and induction of these cytokines by lipopolysaccharide (LPS) was significantly lower in patients than in controls during the first week. Minimum LPS-induced cytokine production correlated strongly with mRS and BI, and also with plasma cortisol.

Conclusion

Absence of spontaneous whole blood gene activation or cytokine production suggests that peripheral blood cells are not the source of cytokines measured in plasma after AIS. Increased plasma IL-1ra within 12 h of AIS onset, the relationship between sTNF-RI and stroke severity, and suppressed cytokine induction suggests early activation of endogenous immunosuppressive mechanisms after AIS.     

The capsule polysaccharide structure and biogenesis for non-O1 Vibrio cholerae NRT36S: genes are embedded in the LPS region

Background  

In V. cholerae, the biogenesis of capsule polysaccharide is poorly understood. The elucidation of capsule structure and biogenesis is critical to understanding the evolution of surface polysaccharide and the internal relationship between the capsule and LPS in this species. V. cholerae serogroup O31 NRT36S, a human pathogen that produces a heat-stable enterotoxin (NAG-ST), is encapsulated. Here, we report the covalent structure and studies of the biogenesis of the capsule in V. cholerae NRT36S.     

入侵种喜旱莲子草和莲子草的营养生长和光合作用对温度的响应

比较温度对入侵种喜旱莲子草(Alternantheraphiloxeroides(Mart.)Griseb.)和其本土近缘种莲子草(A.sessilis(L.)DC.)的营养生长、叶片光合作用及叶绿素荧光的影响。实验将生长均衡的这两种植物放在不同温度(10℃、15℃、20℃、25℃、30℃)的光照培养箱中处理28d。结果表明,喜旱莲子草营养生长的速率和对温度的响应明显不同于莲子草:前者主茎生长的有效积温明显低于后者,分别是11.6d℃和27.0d℃;而新叶萌发的有效积温高于后者,分别是12.1d℃和6.7d℃。入侵种主茎和叶的发育起点温度都比本土种低,分别是10.4℃、11.0℃和12.8℃、14.9℃,表明喜旱莲子草的发育对低温反应不及莲子草敏感。对两种植物叶片的光合作用和叶绿素荧光的测定结果还表明:入侵种比本土种有较高的最大净光合速率和光饱和点,尤其在25℃时;10-30℃的温度范围内喜旱莲子草的最大光化学效率Fv/Fm没有显著变化,而莲子草在10℃低温条件下Fv/Fm值显著降低。较快的主茎生长速率、较宽的温度适应范围以及较高的光合能力可能使喜旱莲子草比本地种具有更强的竞争力,从而在其入侵过程中起了重要作用。     

Recycling of MUC1 is dependent on its palmitoylation

MUC1 is a mucin-like transmembrane protein expressed on the apical surface of epithelia, where it protects the cell surface. The cytoplasmic domain has numerous sites for phosphorylation and docking of proteins involved in signal transduction. In a previous study, we showed that the cytoplasmic YXXphi motif Y20HPM and the tyrosine-phosphorylated Y60TNP motif are required for MUC1 clathrin-mediated endocytosis through binding AP-2 and Grb2, respectively (Kinlough, C. L., Poland, P. A., Bruns, J. B., Harkleroad, K. L., and Hughey, R. P. (2004) J. Biol. Chem. 279, 53071-53077). Palmitoylation of transmembrane proteins can affect their membrane trafficking, and the MUC1 sequence CQC3RRK at the boundary of the transmembrane and cytoplasmic domains mimics reported site(s) of S-palmitoylation. [3H]Palmitate labeling of Chinese hamster ovary cells expressing MUC1 with mutations in CQC3RRK revealed that MUC1 is dually palmitoylated at the CQC motif independent of RRK. Lack of palmitoylation did not affect the cold detergent solubility profile of a chimera (Tac ectodomain and MUC1 transmembrane and cytoplasmic domains), the rate of chimera delivery to the cell surface, or its half-life. Calculation of rate constants for membrane trafficking of wild-type and mutant Tac-MUC1 indicated that the lack of palmitoylation blocked recycling, but not endocytosis, and caused the chimera to accumulate in a EGFP-Rab11-positive endosomal compartment. Mutations CQC/AQA and Y20N inhibited Tac-MUC1 co-immunoprecipitation with AP-1, although mutant Y20N had reduced rates of both endocytosis and recycling, but a normal subcellular distribution. The double mutant chimera AQA+Y20N had reduced endocytosis and recycling rates and accumulated in EGFP-Rab11-positive endosomes, indicating that palmitoylation is the dominant feature modulating MUC1 recycling from endosomes back to the plasma membrane.     

A novel IL-1 receptor, cloned from B cells by mammalian expression, is expressed in many cell types.

cDNA clones corresponding to an Mr approximately 80,000 receptor (type I receptor) for interleukin-1 (IL-1) have been isolated previously by mammalian expression. Here, we report the use of an improved expression cloning method to isolate human and murine cDNA clones encoding a second type (Mr approximately 60,000) of IL-1 receptor (type II receptor). The mature type II IL-1 receptor consists of (i) a ligand binding portion comprised of three immunoglobulin-like domains; (ii) a single transmembrane region; and (iii) a short cytoplasmic domain of 29 amino acids. This last contrasts with the approximately 215 amino acid cytoplasmic domain of the type I receptor, and suggests that the two IL-1 receptors may interact with different signal transduction pathways. The type II receptor is expressed in a number of different tissues, including both B and T lymphocytes, and can be induced in several cell types by treatment with phorbol ester. Both IL-1 receptors appear to be well conserved in evolution, and map to the same chromosomal location. Like the type I receptor, the human type II IL-1 receptor can bind all three forms of IL-1 (IL-1 alpha, IL-1 beta and IL-1ra). Vaccinia virus contains an open reading frame bearing strong resemblance to the type II IL-1 receptor.     

Estimation of the distribution of blood vessel diameters from the arteriovenous passage of microspheres

We present an estimator of the probability density of capillary diameters based on a simple probabilistic model of microsphere passage through a microvascular bed. With this model, an estimate of the density is obtained as the solution of an integral equation. The density is approximated by a linear combination of B-splines with the coefficients restricted to be nonnegative. Equations for maximum likelihood estimates of the coefficients are obtained. The proposed method is applied to data from the dog intestinal circulation.     

玉米叶片生长部位渗透调节和生长的生物物理参数变化

玉米叶片生长部位随着水分胁迫加剧ψ_w降低、LER减慢。LER从最大到零,快速干旱处理的ψw从-0.55降至-0.85 MPa;缓慢干旱处理ψ_w从-0.88降至-1.13 MPa。在任何一种LER下,缓慢干旱处理的ψ_s比快速干旱处理更低,生长停止时,前者为-1.57 MPa,而后者为-1.30MPa。缓慢干旱叶片尽管在更低ψ_w下,仍能维持一定膨压,保持一定的生长速率。经历长时间水分胁迫会改变细胞延伸生长的生物物理参数,增大临界膨压(0.08～0.09 MPa)。这是水分胁迫植株,在一定ψ_p下生长速率减慢的原因。