This pilot study used an immune thrombocytopenic purpura (ITP)-specific bleeding score, the ITP Bleeding Scale (IBLS) to analyse the correlation of clinical and laboratory platelet variables with bleeding. MethodsA prospective Institutional Review Board-approved study was conducted on 100 visits for 65 consenting patients with ITP (30 on one and 35 on two visits) from December 2004 to December 2005. The patients were primarily adults with chronic ITP; a minority were children or adults with acute ITP.The IBLS (Table I) comprised 11 grades from 0 (none) to 2 (marked bleeding) assessed at nine anatomical sites by history over the previous week (Hx). In addition, two of these sites, skin and oral, were also assessed by physical examination (PE). The 'worst ever' bleeding experienced at each site was graded using the same system. Blood counts were analysed by using the Bayer-ADVIA TM 120 (Giacomini et al, 2001). Large platelets (20-60 fl) were examined as they were reported to be more haemostatically active (Karpatkin, 1978;Michel et al, 2005). Analysis focused on the six grades with most bleeding -skin and oral (Hx and PE), epistaxis and gynaecological (GYN) (Hx). Insufficient haemorrhages occurred at the other sites to enable inclusion in the analysis. Means, medians, ranges, the Kruskal-Wallis, Fisher's exact and chi-squared tests, and the Kappa statistic to establish inter-observer reliability (for 63/100 visits) were calculated. P-values <0AE05 were considered significant. ResultsThe median age was 31AE5 years; 20 patients were under 18 and eight over 65 years. Forty-two were female, 23 of reproductive age and assessed for GYN bleeding on 36 visits. Eighty-eight per cent had chronic ITP (median duration 6 years) and 40% were splenectomised. No correlation between IBLS and age, sex, duration of ITP or splenectomy status was found.The IBLS grades on the 100 study visits are presented in Tables II-IV. Seventy-two per cent of grade 1 haemorrhages occured in the skin. Grade 2 bleeding was more heterogeneous (28% skin, 46% oral, 13% GYN and 10% epistaxis). Patients with more skin bleeding also had more oral bleeding (PE and Hx, all P-values <0AE026).The median platelet count for all visits was 39AE5 · 10 9 /l (range: 6-623 · 10 9 /l). Twenty-five visits had a platelet count £ 20 · 10 9 /l and 46 visits had a platelet SummaryA method for objective quantification of bleeding symptoms in immune thrombocytopenic purpura (ITP) has not been established. The ITP Bleeding Scale (IBLS) is a novel bleeding assessment system comprising 11 site-specific grades. Implementation of the IBLS on 100 patient visits revealed that although platelet count and large platelet count correlated well with bleeding symptoms overall, this relationship disappeared in marked thrombocytopenia. The IBLS is a useful clinical tool for monitoring bleeding and may be used to aid the development of laboratory parameters that correlate with underlying bleeding propensity in thrombocytopenia.

Community differentiation is a fundamental topic of the social sciences, and its prehistoric origins in Europe are typically assumed to lie among the complex, densely populated societies that developed millennia after their Neolithic predecessors. Here we present the earliest, statistically significant evidence for such differentiation among the first farmers of Neolithic Europe. By using strontium isotopic data from more than 300 early Neolithic human skeletons, we find significantly less variance in geographic signatures among males than we find among females, and less variance among burials with ground stone adzes than burials without such adzes. From this, in context with other available evidence, we infer differential land use in early Neolithic central Europe within a patrilocal kinship system.

A proteomic approach has been applied to investigate changes in the extracellular matrix of Arabidopsis thaliana cell suspension cultures following treatments with two fungal pathogen elicitors, chitosan and extracts of Fusarium moniliforme. The oxidative burst and induction of glutathione S-transferase were used as markers for induction of the pathogen defence response. Changes in the cell wall and culture filtrate proteome were profiled. Proteins whose abundance changed reproducibly were analysed via matrix assisted laser desorption ionisation-time of flight (MALDI-TOF) mass spectrometry (MS). An increase in the level of two classical cell wall proteins (a putative endochitinase and a polygalacturonase inhibiting protein) and two novel proteins (a putative receptor-like protein kinase and a probable apospory-associated protein) were seen at 24 hours following elicitation. The level of an unknown protein and a hypothetical protein, which has some homology to serine carboxypeptidases, were decreased at 24 hours post-elicitation. In the culture filtrate extracts, we identified two pathogen elicitor responsive proteins, a xyloglucan endo-1,4-beta-D glucanases (XEG) and a peroxidase. Using a combination of two-dimensional polyacrylamide gel electrophoresis, immunoblotting with a phosphotyrosine-specific antibody, and MALDI-TOF MS we discovered that spots that represent putative lectin receptor-like kinase, a putative endochitinase and a XEG possess phosphorylated tyrosine residues. The identification of phosphorylated bona fide cell wall proteins and a putative extracellular receptor-like kinase with no transmembrane domain implicate the existence of an extracellular phosphorylation network which could be involved in intercellular communication.

SUMMARYIn healthy plants extracellular ATP (eATP) regulates the balance between cell viability and death. Here we show an unexpected critical regulatory role of eATP in disease resistance and defensive signalling. In tobacco, enzymatic depletion of eATP or competition with non-hydrolysable ATP analogues induced pathogenesisrelated (PR) gene expression and enhanced resistance to tobacco mosaic virus and Pseudomonas syringae pv. tabaci. Artificially increasing eATP concentrations triggered a drop in levels of the important defensive signal chemical salicylic acid (SA) and compromised basal resistance to viral and bacterial infection. Inoculating tobacco leaf tissues with bacterial pathogens capable of activating PR gene expression triggered a rapid decline in eATP. Conversely, inoculations with mutant bacteria unable to induce defence gene expression failed to deplete eATP. Furthermore, a collapse in eATP concentration immediately preceded PR gene induction by SA. Our study reveals a previously unsuspected role for eATP as a negative regulator of defensive signal transduction and demonstrates its importance as a key signal integrating defence and cell viability in plants.