Long-term outcomes of the randomized controlled trial comparing 5-aminolaevulinic acid and Photofrin photodynamic therapy for Barrett's oesophagus related neoplasia

OBJECTIVE: Photodynamic therapy (PDT) was used as therapy for early neoplasia associated with Barrett's oesophagus (BE). This is 5-year follow-up of patients enrolled into randomised controlled trial of 5-aminolaevulinic acid (ALA) vs. Photofrin PDT. METHODS: Biopsies were taken from original Barrett's segment during endoscopic follow up using Seattle protocol. Endoscopic mucosal resection (EMR) ± radiofrequency ablation (RFA) was preferred therapy in patients who failed PDT and/or had recurrent neoplasia. RESULTS: Fifty eight of 64 patients enrolled in the original trial were followed up including 31 patients treated with ALA PDT (17 patients with ≤6 cm, 14 patients with >6 cm segment of BE) and 27 treated with Photofrin PDT (14 patients with ≤6 cm, 13 patients with >6 cm BE). Initial success was achieved in 65% (20/31) ALA and 48% (13/27) Photofrin patients (p = .289). Thirty five percent patients (7/20) relapsed in ALA group and 54% (7/13) relapsed in Photofrin group (p = .472). At a median follow-up of 67 months, no significant difference was found in long-term complete reversal of intestinal metaplasia (CR-IM) and complete reversal of dysplasia (CR-D) between ALA and Photofrin groups (78% vs. 63%; p = .18; 90% vs. 76%; p = .26). Original length of BE did not alter long-term outcome. Four patients from each group progressed to invasive oesophageal adenocarcinoma. Initial success of ALA PDT was associated with significantly better likelihood of long-term remission (p = .03). CONCLUSIONS: Initial response to PDT plays key role in long term outcome. RFA ± EMR have, however, become preferred minimally invasive ablative therapy for BE-related neoplasia due to poor efficacy of PDT

Age, puberty and attractiveness judgments in adolescents

Previous work has suggested that judgments of the attractiveness of some facial and vocal features change during adolescence. Here, over 70 Czech adolescents aged 12–14 made forced-choice attractivenessjudgments on adolescent faces manipulated in symmetry, averageness and femininity, and on adolescent opposite-sex voices manipulated in fundamental frequency (perceived as pitch), and completed questionnaires on pubertal development. Consistent with typical adult judgments, adolescents selected the symmetric, average and feminine male and female faces as more attractive significantly more often than the asymmetric, non-average and masculine faces respectively. Moreover, preferences for symmetric faces were positively associated with adolescents’ age and stage of pubertal development. Unexpectedly, voice pitch did not significantly influence adolescents’ attractivenessjudgments. Collectively, these findings present new evidence using refined methodology that adolescent development is related to variation in attractivenessjudgments

Novel APC mutations in Czech and Slovak FAP families: clinical and genetic aspects

BACKGROUND: Germline mutations in the adenomatous polyposis gene (APC) result in familial adenomatous polyposis (FAP). FAP is an autosomal dominantly inherited disorder predisposing to colorectal cancer. Typical FAP is characterized by hundreds to thousands of colorectal adenomatous polyps and by several extracolonic manifestations. An attenuated form of polyposis (AFAP) is characterized by less than 100 adenomas and later onset of the disease. METHODS: Here, we analyzed the APC gene for germline mutations in 59 Czech and 15 Slovak FAP patients. In addition, 50 apparently APC mutation negative Czech probands and 3 probands of Slovak origin were screened for large deletions encompassing the APC gene. Mutation screening was performed using denaturing gradient gel electrophoresis and/or protein truncation test. DNA fragments showing an aberrant electrophoretic banding pattern were sequenced. Screening for large deletions was performed by multiplex ligation dependent probe amplification. The extent of deletions was analyzed using following microsatellite markers: D5S299, D5S82, D5S134 and D5S346. RESULTS: In the set of Czech and Slovak patients, we identified 46 germline mutations among 74 unrelated probands. Total mutation capture is 62,2% including large deletions. Thirty seven mutations were detected in 49 patients presenting a classical FAP phenotype (75,5%) and 9 mutations in 25 patients with attenuated FAP (36%). We report 20 novel germline APC mutations and 3 large deletions (6%) encompassing the whole-gene deletions and/or exon 14 deletion. In the patients with novel mutations, correlations of the mutation localization are discussed in context of the classical and/or attenuated phenotype of the disease. CONCLUSION: The results of the molecular genetic testing are used both in the establishment of the predictive diagnosis and in the clinical management of patients. In some cases this study has also shown the difficulty to classify clinically between the classical and the attenuated form of FAP according to the established criteria. Interfamilial and/or intrafamilial phenotype variability was also confirmed in some cases which did not fit well with predicted genotype-phenotype correlation. All these findings have to be taken into consideration both in the genetic counselling and in the patient care

Inactivation of promoter 1B of APC causes partial gene silencing: evidence for a significant role of the promoter in regulation and causative of familial adenomatous polyposis

Familial adenomatous polyposis (FAP) is caused by germline mutations in the adenomatous polyposis coli (APC) gene. Two promoters, 1A and 1B, have been recognized in APC, and 1B is thought to have a minor role in the regulation of the gene. We have identified a novel deletion encompassing half of this promoter in the largest family (Family 1) of the Swedish Polyposis Registry. The mutation leads to an imbalance in allele-specific expression of APC, and transcription from promoter 1B was highly impaired in both normal colorectal mucosa and blood from mutation carriers. To establish the significance of promoter 1B in normal colorectal mucosa (from controls), expression levels of specific transcripts from each of the promoters, 1A and 1B, were examined, and the expression from 1B was significantly higher compared with 1A. Significant amounts of transcripts generated from promoter 1B were also determined in a panel of 20 various normal tissues examined. In FAP-related tumors, the APC germline mutation is proposed to dictate the second hit. Mutations leaving two or three out of seven 20-amino-acid repeats in the central domain of APC intact seem to be required for tumorigenesis. We examined adenomas from mutation carriers in Family 1 for second hits in the entire gene without any findings, however, loss of the residual expression of the deleterious allele was observed. Three major conclusions of significant importance in relation to the function of APC can be drawn from this study; (i) germline inactivation of promoter 1B is disease causing in FAP; (ii) expression of transcripts from promoter 1B is generated at considerable higher levels compared with 1A, demonstrating a hitherto unknown importance of 1B; (iii) adenoma formation in FAP, caused by impaired function of promoter 1B, does not require homozygous inactivation of APC allowing for alternative genetic models as basis for adenoma formation

Molecular Analysis of the APC and MYH Genes in Czech Families Affected by FAP or Multiple Adenomas: 13 Novel Mutations

We give a nearly optimal sublinear-time algorithm for approximating the size of a minimum vertex cover in a graph G. The algorithm may query the degree deg(v) of any vertex v of its choice, and for each 1 <= i <= deg(v), it may ask for the i-th neighbor of v. Letting VC_opt(G) denote the minimum size of vertex cover in G, the algorithm outputs, with high constant success probability, an estimate VC_estimate(G) such that VC_opt(G) <= VC_estimate(G) <= 2 * VC_opt(G) + epsilon*n, where epsilon is a given additive approximation parameter. We refer to such an estimate as a (2,epsilon)-estimate. The query complexity and running time of the algorithm are ~O(avg_deg * poly(1/epsilon)), where avg_deg denotes the average vertex degree in the graph. The best previously known sublinear algorithm, of Yoshida et al. (STOC 2009), has query complexity and running time O(d^4/epsilon^2), where d is the maximum degree in the graph. Given the lower bound of Omega(avg_deg) (for constant epsilon) for obtaining such an estimate (with any constant multiplicative factor) due to Parnas and Ron (TCS 2007), our result is nearly optimal. In the case that the graph is dense, that is, the number of edges is Theta(n^2), we consider another model, in which the algorithm may ask, for any pair of vertices u and v, whether there is an edge between u and v. We show how to adapt the algorithm that uses neighbor queries to this model and obtain an algorithm that outputs a (2,epsilon)-estimate of the size of a minimum vertex cover whose query complexity and running time are ~O(n) * poly(1/epsilon)

Crystallization and preliminary crystallographic characterization of the extrinsic PsbP protein of photosystem II from Spinacia oleracea

Degradation-free crystalization of thrombin-digested recombinant His-tagged PsbP protein of photosystem II from Spinacia oleracea resulting in crystals diffracting to 2.06 Å

Air-tolerant C-C bond formation via organometallic ruthenium catalysis: diverse catalytic pathways involving (C5Me5)Ru or (C5H5)Ru are robust to molecular oxygen

Ruthenium-catalyzed substitutions of carbon pronucleophiles, various 2+2+2 cycloadditions, and addition of a diazo compound to an alkyne are shown to proceed in the presence of air. Notably diverse catalytic manifolds remain supported under conditions generally regarded as prohibitive. Building on rare reports from the literature we show that a range of organometallic transformations based on reaction intermediates derived from (C5Me5)Ru or (C5H5)Ru moieties are air-compatible. © 2009 Elsevier Ltd. All rights reserved