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The Spanish team of the Human Proteome Project (SpHPP) marked the annotation of Chr16 and data analysis as one of its priorities. Precise annotation of Chromosome 16 proteins according to C-HPP criteria is presented. Moreover, Human Body Map 2.0 RNA-Seq and Encyclopedia of DNA Elements (ENCODE) data sets were used to obtain further information relative to cell/tissue specific chromosome 16 coding gene expression patterns and to infer the presence of missing proteins. Twenty-four shotgun 2D-LC-MS/MS and gel/LC-MS/MS MIAPE compliant experiments, representing 41% coverage of chromosome 16 proteins, were performed. Furthermore, mapping of large-scale multicenter mass spectrometry data sets from CCD18, MCF7, Jurkat, and Ramos cell lines into RNA-Seq data allowed further insights relative to correlation of chromosome 16 transcripts and proteins. Detection and quantification of chromosome 16 proteins in biological matrices by SRM procedures are also primary goals of the SpHPP. Two strategies were undertaken: one focused on known proteins, taking advantage of MS data already available, and the second, aimed at the detection of the missing proteins, is based on the expression of recombinant proteins to gather MS information and optimize SRM methods that will be used in real biological samples. SRM methods for 49 known proteins and for recombinant forms of 24 missing proteins are reported in this study.

Background: No validated instruments have been developed to measure work disability in Crohn's disease (CD). The aim of our study was to develop and validate a CD perceived work disability questionnaire (CPWDQ)., Methods: Development phase: an initial questionnaire containing 52 items was obtained from patients' interviews plus additional sources; it was completed by 106 patients and the 16 most significant items were selected using a psychometric method in order to create the CPWDQ. Validation phase: The validation assessed the questionnaire's convergent validity, discriminant validity, test-retest reproducibility, and internal consistency in 108 patients. Spearman rank correlation, t-test, intraclass correlation, and Cronbach's alpha were used for the analysis., Results: Convergent validity was confirmed by good correlations between the CPWDQ and: clinical activity (r = 0.59, P < 0.01), the Short Inflammatory Bowel Disease Questionnaire, IBDQ-9, (r = 0.76, P < 0.001), Euroqol-5D (r = 0.53, P < 0.01), and overall work impairment (WPAI&#95;CD) r = 0.66 (P < 0.01). Discriminant validity: CPWQ scores were higher in patients expected to have more severe disability, that is, in patients with active disease (n = 38) 32.3 ± 7.3 versus inactive (n = 70) 22.6 ± 5.9 (P < 0.001), in those requiring previous sick leave 30.7 ± 7.5 (n = 45) versus no sick leave 22.6 ± 6.6 (n = 63) (P < 0.01), and in those requiring hospitalization 32.2 ± 8.6 (n = 18) versus no hospitalization 24.7 ± 7.1 (n = 90) (P < 0.01). Internal consistency was also good (Cronbach's alpha = 0.89). Reproducibility: CPWDQ measures obtained 2 weeks apart showed an excellent intraclass correlation coefficient: 0.89 (95% CI: 0.83-0.93)., Conclusions: The CPWDQ seems to be a valid, reliable tool for measuring subjective work disability in CD., (Copyright © 2011 Crohn's & Colitis Foundation of America, Inc.)

Resistance to anti-retroviral therapy is one of the main problems in the favorable outcome of treatment in HIV patients, as well as toxicity and adherence to treatment. Resistance has increased in recent years, and it is evaluated through genotyping and phenotypic tests. Information provided by these studies is crucial when deciding the most appropriate treatment. However, genotype interpretation is complex and subject to frequent change, because of the incorporation of new drugs and the appearance of new resistance patterns. This review aims ,understanding the fundamental concepts of antiretroviral resistance (ARV), which examines the general principles, mechanisms and patterns of resistance, both for the traditional family of anti-retrovirals, as well as for the most recently licensed drugs.

Objectives: The Work Productivity and Activity Impairment Questionnaire (WPAI) is an instrument that measures work impairment in the setting of different diseases. One previous study validating the WPAI in Crohn's disease (CD) patients suggested that its reproducibility may be unsatisfactory. This study evaluated the validity and reproducibility of the Spain Spanish version of the WPAI questionnaire in CD patients., Methods: In a prospective study, convergent validity was tested comparing WPAI with quality of life and CD activity indices. Known-groups validity was assessed by comparing WPAI scores in CD patients with active versus inactive disease. Reproducibility of repeated tests was evaluated by comparing two WPAI measures obtained two to four weeks apart in stable patients., Results: One hundred eight patients were included in the various steps of the analysis. The WPAI showed adequate known groups validity and convergent validity. The reproducibility of the test and, specifically, that of the evaluation of presenteeism (the impairment of productivity while working) were good., Conclusions: This study confirms the validity of WPAI for measuring work impairment in CD patients. In contrast to previous studies, the test reproducibility was adequate., (Copyright © 2011 International Society for Pharmacoeconomics and Outcomes Research (ISPOR). Published by Elsevier Inc. All rights reserved.)

Modified fibres with high-cellulose content were obtained with two new bacterial xylanases from families 11 and 5. These xylanases were applied separately or simultaneously in a complete ECF (Elemental Chlorine Free) bleaching sequence. Both xylanases improved delignification and bleaching during the sequence and a synergistic effect of the enzymes was observed on several pulp and paper properties. The xylanases boosted the release of xylooligosaccharides branched with hexenuronic acids (HexA), giving rise to fibres with a reduced HexA and xylose content. However, these effects depended on the xylanase used, being the family 11 enzyme more efficient than the family 5 xylanase. Effluent properties such as absorbance spectra UV/Vis, COD and colour were affected by the enzymatic sequences as a consequence of the dissolution of lignin and xylooligosaccharides. Some changes in the fibre morphology were also produced without affecting the final paper strength properties., (Copyright 2010 Elsevier Ltd. All rights reserved.)

A new bacterial xylanase belonging to family 5 of glycosyl hydrolases was identified and characterized. The xylanase, Xyn5B from Bacillus sp. strain BP-7, was active on neutral, nonsubstituted xylooligosaccharides, showing a clear difference from other GH5 xylanases characterized to date that show a requirement for methyl-glucuronic acid side chains for catalysis. The enzyme was evaluated on Eucalyptus kraft pulp, showing its effectiveness as a bleaching aid.

Paenibacillus barcinonensis is a soil bacterium bearing a complex set of enzymes for xylan degradation, including several secreted enzymes and Xyn10B, one of the few intracellular xylanases reported to date. The crystal structure of Xyn10B has been determined by x-ray analysis. The enzyme folds into the typical (beta/alpha)(8) barrel of family 10 glycosyl hydrolases (GH10), with additional secondary structure elements within the beta/alpha motifs. One of these loops -L7- located at the beta7 C terminus, was essential for xylanase activity as its partial deletion yielded an inactive enzyme. The loop contains residues His(249)-Glu(250), which shape a pocket opened to solvent in close proximity to the +2 subsite, which has not been described in other GH10 enzymes. This wide cavity at the +2 subsite, where methyl-2,4-pentanediol from the crystallization medium was found, is a noteworthy feature of Xyn10B, as compared with the narrow crevice described for other GH10 xylanases. Docking analysis showed that this open cavity can accommodate glucuronic acid decorations of xylo-oligosaccharides. Co-crystallization experiments with conduramine derivative inhibitors supported the importance of this open cavity at the +2 subsite for Xyn10B activity. Several mutant derivatives of Xyn10B with improved thermal stability were obtained by forced evolution. Among them, mutant xylanases S15L and M93V showed increased half-life, whereas the double mutant S15L/M93V exhibited a further increase in stability, showing a 20-fold higher heat resistance than the wild type xylanase. All the mutations obtained were located on the surface of Xyn10B. Replacement of a Ser by a Leu residue in mutant xylanase S15L can increase hydrophobic packing efficiency and fill a superficial indentation of the protein, giving rise to a more compact structure of the enzyme.

Background & Aims: Chronic disability and its consequences for social life and employment are important but often neglected aspects of Crohn's disease. No specific scores have been developed to evaluate chronic disability in patients with Crohn's disease; the medical criteria used by government authorities to award disability benefits have not been analyzed. We aimed to determine the courts' criteria for awarding disability benefits to patients with Crohn's disease in Spain., Methods: We systematically searched case law databases in Spain's regional Supreme Courts to identify sentences regarding awards of disability benefits to patients with Crohn's disease. Selected decisions were reviewed to extract variables related to the awarding of benefits. Univariate and multivariate analyses were performed to determine which variables predicted the awarding of benefits., Results: Two hundred eighty sentences were reviewed. The rate of judicial decisions in favor of the claimants varied considerably between the various tribunals. Multivariate analysis showed that adequate description of the disease (odds ratio, 8.6), fecal incontinence (odds ratio, 8.9), the number of associated diseases (odds ratio, 2.3), and the presence of an ostomy (odds ratio, not estimable) were independent predictors of the awarding of Social Security benefits., Conclusions: The amount of Social Security benefits awarded to patients with Crohn's disease varied depending on the tribunal. The most important predictors of a court's disability award were the adequate description of the patient's disease, fecal incontinence, associated diseases, and presence of an ostomy.

Background: Immunotherapy can be used to treat allergic reactions to insect stings, specially bees and wasps., Aim: To report the experience with immunotherapy with aqueous extracts of hymenoptera venoms (bees and wasps)., Material and Methods: Ten patients aged 6 to 58 years were treated in an allergy center of a University Clinical Hospital. The medical indication for this treatment was, in all patients, anaphylactic reactions after hymenoptera stings. Immunotherapy was carried out using standardized vaccines (Aqueous extracts Venomvac LETI, Spain), applied in a traditional protocol, with subcutaneous injections. This protocol had two phases: a buildup phase (between weeks 1 and 13) and a monthly maintenance phase, from the 13th week. The monthly maintenance dose was 100 fig of hymenoptera specific venom extract., Results: Six patients had adverse reactions of different severity, during the treatment protocols and all had a good response to immediate therapeutic measures. After these events, they followed the protocol without problems. Two patients, treated with bee vaccines, suffered an accidental bee sting during the maintenance phase and they developed only local reactions., Conclusions: The lack of adverse reactions to bee stings in these two patients indicates the acquisition of clinical tolerance.

Background: Thiopurinic immunomodulators are effective for maintaining symptom remission in Crohn's disease. Little is known, however, about their effect on patients' quality of life or psychological well-being. The present study aimed to determine whether remission induced by thiopurinic immunomodulators returns levels of quality of life and psychological well-being to normal., Materials and Methods: A case-control study was performed. Cases were 33 patients with Crohn's disease treated with azathioprine or 6-mercaptopurine and in stable remission for at least 6 months. Sixty-six healthy individuals matched 2:1 by age and sex and 14 patients with active Crohn's disease were included as control groups. Quality of life was evaluated with the Short Form (SF-36) questionnaire, and the respective Hamilton rating scales were used for anxiety and depression. ANOVA with Bonferroni's correction was used for multiple comparisons., Results: SF-36 global scores were 85 in the study group, 85 in healthy controls (P = 1), and 58.6 in patients with active disease (P < 0.001 for the comparison with the other 2 groups). The differences between values were 0 (95% CI -4-4), 26.4 (95% CI 20-32), and 26.4 (95% CI 19-33), respectively. The respective anxiety and depression scores were 6.5, 5.5, and 16.2 and 3.7, 3.3, and 10.9. No significant differences were observed in any of the SF-36 domains between case and control groups, whereas in patients with active disease, all domains were significantly worse., Conclusions: Thiopurinic immunomodulator-induced remission restores normal levels of quality of life and psychological well-being in Crohn's disease patients.

Xylanase A from Bacillus sp. BP7, an enzyme with potential applications in biotechnology, was used to test Pir4, a disulfide bound cell wall protein, as a fusion partner for the expression of recombinant proteins in standard or glycosylation-deficient mnn9 strains of Saccharomyces cerevisiae. Five different constructions were carried out, inserting in-frame the coding sequence of xynA gene in that of PIR4, with or without the loss of specific regions of PIR4. Targeting of the xylanase fusion protein to the cell wall was achieved in two of the five constructions, while secretion to the growth medium was the fate of the gene product of one of the constructions. In all three cases localization of the xylanase fusion proteins was confirmed both by Western blot and detection with Pir-specific antibodies and by xylanase activity determination. The cell wall-targeted fusion proteins could be extracted by reducing agents, showing that the inclusion of a recombinant protein of moderate size does not affect the way Pir4 is attached to the cell wall. Also, the construction that leads to the secretion of the fusion protein permitted us to identify a region of Pir4 responsible for cell wall retention. In summary, we have developed a Pir4-based system that allows selective targeting of an active recombinant enzyme to the cell wall or the growth medium. This system may be of general application for the expression of heterologous proteins in S. cerevisiae for surface display and secretion.

The xynA gene encoding a xylanase from the recently isolated Bacillus sp. strain BP-7 has been cloned and expressed in Escherichia coli. Recombinant xylanase A showed high activity on xylans from hardwoods and cereals, and exhibited maximum activity at pH 6 and 60 degrees C. The enzyme remained stable after incubation at 50 degrees C and pH 7 for 3 h, and it was strongly inhibited by Mn(2+), Fe(3+), Pb(2+), and Hg(2+). Analysis of xylanase A in zymograms showed an apparent molecular size of 24 kDa and a pI of above 9. The amino acid sequence of xylanase A, as deduced from xynA gene, shows homology to alkaline pI-low molecular weight xylanases of family 11 such as XynA from Bacillus subtilis. Analysis of codon usage in xynA from Bacillus sp. BP-7 shows that the G+C content at the first and second codon positions is notably different from the mean values found for glycosyl hydrolase genes from Bacillus subtilis.