Enzymatic reduction of azo and indigoid compounds

A customer- and environment-friendly method for the decolorization azo dyes was developed. Azoreductases could be used both to bleach hair dyed with azo dyes and to reduce dyes in vat dyeing of textiles. A new reduced nicotinamide adenine dinucleotide-dependent azoreductase of Bacillus cereus, which showed high potential for reduction of these dyes, was purified using a combination of ammonium sulfate precipitation and chromatography and had a molecular mass of 21.5 kDa. The optimum pH of the azoreductase depended on the substrate and was within the range of pH 6 to 7, while the maximum temperature was reached at 40°C. Oxygen was shown to be an alternative electron acceptor to azo compounds and must therefore be excluded during enzymatic dye reduction. Biotransformation of the azo dyes Flame Orange and Ruby Red was studied in more detail using UV-visible spectroscopy, high-performance liquid chromatography, and mass spectrometry (MS). Reduction of the azo bonds leads to cleavage of the dyes resulting in the cleavage product 2-amino-1,3 dimethylimidazolium and N∼1∼,N∼1∼-dimethyl-1,4-benzenediamine for Ruby Red, while only the first was detected for Flame Orange because of MS instability of the expected 1,4-benzenediamine. The azoreductase was also found to reduce vat dyes like Indigo Carmine (C.I. Acid Blue 74). Hydrogen peroxide (H2O2) as an oxidizing agent was used to reoxidize the dye into the initial form. The reduction and oxidation mechanism of Indigo Carmine was studied using UV-visible spectroscopy

Foreword

The postsynaptic density protein-95/disks large/zonula occludens-1 (PDZ) protein domain family is one of the most common proteinprotein interaction modules in mammalian cells, with paralogs present in several hundred human proteins. PDZ domains are found in most cell types, but neuronal proteins, for example, are particularly rich in these domains. The general function of PDZ domains is to bring proteins together within the appropriate cellular compartment, thereby facilitating scaffolding, signaling, and trafficking events. The many functions of PDZ domains under normal physiological as well as pathological conditions have been reviewed recently. In this review, we focus on the molecular details of how PDZ domains bind their protein ligands and their potential as drug targets in this context

Optical BVI Imaging and HI Synthesis Observations of the Dwarf Irregular Galaxy ESO 364-G 029

As part of an effort to enlarge the number of well-studied Magellanic-type galaxies, we obtained broadband optical imaging and neutral hydrogen radio synthesis observations of the dwarf irregular galaxy ESO 364-G 029. The optical morphology characteristically shows a bar-like main body with a one-sided spiral arm, an approximately exponential light distribution, and offset photometric and kinematic centers. The HI distribution is mildly asymmetric and, although slightly offset from the photometric center, roughly follows the optical brightness distribution, extending to over 1.2 Holmberg radii (where mu_B = 26.5 mag/arcsec^2). In particular, the highest HI column densities closely follow the bar, one-arm spiral, and a third optical extension. The rotation is solid-body in the inner parts but flattens outside of the optical extent. The total HI flux F_HI = 23.1 pm 1.2 Jy km/s, yielding a total HI mass M_HI= (6.4 pm 1.7) x 10^8 Msun (for a distance D = 10.8 pm 1.4 Mpc) and a total HI mass-to-blue-luminosity ratio M_HI/L_B = (0.96 pm 0.14) Msun / Lsun,B (distance independent). The HI data suggest a very complex small-scale HI structure, with evidence of large shells and/or holes, but deeper observations are required for a detailed study. Follow-up observations are also desirable for a proper comparison with the Large Magellanic Cloud, where despite an optical morphology very similar to ESO 364-G 029 the HI bears little resemblance to the optical.Comment: 10 pages, 11 figures, accepted by A&

A Constrained Fuzzy Knowledge-Based System for the Management of Container Yard Operations

The management of container yard operations is considered by yard operators to be a very challenging task due to the many uncertainties inherent in such operations. The storage of the containers is one of those operations that require proper management for the efficient utilisation of the yard, requiring rapid retrieval time and a minimum number of re-handlings. The main challenge is when containers of a different size, type, or weight need to be stored in a yard that holds a number of pre-existing containers. This challenge becomes even more complex when the date and time for the departure of the containers are unknown, as is the case when the container is collected by a third-party logistics company without any prior notice being given. The aim of this study is to develop a new system for the management of container yard operations that takes into consideration a number of factors and constraints that occur in a real-life situation. One of these factors is the duration of stay for the topmost containers of each stack, when the containers are stored. Because the duration of stay for containers in a yard varies dynamically over time, an ‘ON/OFF’ strategy is proposed to activate/deactivate the duration of stay factor constraint if the length of stay for these containers varies significantly over time. A number of tools and techniques are utilised for developing the proposed system including: discrete event simulation for the modelling of container storage and retrieval operations, a fuzzy know ledge-based model for the stack allocation of containers, and a heuristic algorithm called ‘neighbourhood’ for the container retrieval operation. Results show that by adopting the proposed ‘ON/OFF’ strategy, 5% of the number of re-handlings, 2.5% of the total retrieval time, 6.6% of the total re-handling time and 42% of the average waiting time per truck are reduced

Recommendations for reporting results of diagnostic genetic testing (biochemical, cytogenetic and molecular genetic)

Genetic test results can have considerable importance for patients, their parents and more remote family members. Clinical therapy and surveillance, reproductive decisions and genetic diagnostics in family members, including prenatal diagnosis, are based on these results. The genetic test report should therefore provide a clear, concise, accurate, fully interpretative and authoritative answer to the clinical question. The need for harmonizing reporting practice of genetic tests has been recognised by the External Quality Assessment (EQA), providers and laboratories. The ESHG Genetic Services Quality Committee has produced reporting guidelines for the genetic disciplines (biochemical, cytogenetic and molecular genetic). These guidelines give assistance on report content, including the interpretation of results. Selected examples of genetic test reports for all three disciplines are provided in an annexe.</p

53BP1 can limit sister-chromatid rupture and rearrangements driven by a distinct ultrafine DNA bridging-breakage process

Chromosome missegregation acts as one of the driving forces for chromosome instability and cancer development. Here, we find that in human cancer cells, HeLa and U2OS, depletion of 53BP1 (p53-binding protein 1) exacerbates chromosome non-disjunction resulting from a new type of sister-chromatid intertwinement, which is distinct from FANCD2-associated ultrafine DNA bridges (UFBs) induced by replication stress. Importantly, the sister DNA intertwinements trigger gross chromosomal rearrangements through a distinct process, named sister-chromatid rupture and bridging. In contrast to conventional anaphase bridge-breakage models, we demonstrate that chromatid axes of the intertwined sister-chromatids rupture prior to the breakage of the DNA bridges. Consequently, the ruptured sister arms remain tethered and cause signature chromosome rearrangements, including whole-arm (Robertsonian-like) translocation/deletion and isochromosome formation. Therefore, our study reveals a hitherto unreported chromatid damage phenomenon mediated by sister DNA intertwinements that may help to explain the development of complex karyotypes in tumour cells

Identification of a Common Gene Expression Response in Different Lung Inflammatory Diseases in Rodents and Macaques

To identify gene expression responses common to multiple pulmonary diseases we collected microarray data for acute lung inflammation models from 12 studies and used these in a meta-analysis. The data used include exposures to air pollutants; bacterial, viral, and parasitic infections; and allergic asthma models. Hierarchical clustering revealed a cluster of 383 up-regulated genes with a common response. This cluster contained five subsets, each characterized by more specific functions such as inflammatory response, interferon-induced genes, immune signaling, or cell proliferation. Of these subsets, the inflammatory response was common to all models, interferon-induced responses were more pronounced in bacterial and viral models, and a cell division response was more prominent in parasitic and allergic models. A common cluster containing 157 moderately down-regulated genes was associated with the effects of tissue damage. Responses to influenza in macaques were weaker than in mice, reflecting differences in the degree of lung inflammation and/or virus replication. The existence of a common cluster shows that in vivo lung inflammation in response to various pathogens or exposures proceeds through shared molecular mechanisms

Differential gene expression profile reveals deregulation of pregnancy specific β1 glycoprotein 9 early during colorectal carcinogenesis

BACKGROUND: APC (Adenomatous polyposis coli) plays an important role in the pathogenesis of both familial and sporadic colorectal cancer. Patients carrying germline APC mutations develop multiple colonic adenomas at younger age and higher frequency than non-carrier cases which indicates that silencing of one APC allele may be sufficient to initiate the transformation process. METHODS: To elucidate the biological dysregulation underlying adenoma formation we examined global gene expression profiles of adenomas and corresponding normal mucosa from an FAP patient. Differential expression of the most significant gene identified in this study was further validated by mRNA in situ hybridization, reverse transcriptase PCR and Northern blotting in different sets of adenomas, tumours and cancer cell lines. RESULTS: Eighty four genes were differentially expressed between all adenomas and corresponding normal mucosa, while only seven genes showed differential expression within the adenomas. The first group included pregnancy specific β-1 glycoprotein 9 (PSG9) (p < 0.006). PSG9 is a member of the carcinoembryonic antigen (CEA)/PSG family and is produced at high levels during pregnancy, mainly by syncytiotrophoblasts. Further analysis of sporadic and familial colorectal cancer confirmed that PSG9 is ectopically upregulated in vivo by cancer cells. In total, deregulation of PSG9 mRNA was detected in 78% (14/18) of FAP adenomas and 75% (45/60) of sporadic colorectal cancer cases tested. CONCLUSION: Detection of PSG9 expression in adenomas, and at higher levels in FAP cases, indicates that germline APC mutations and defects in Wnt signalling modulate PSG9 expression. Since PSG9 is not found in the non-pregnant adult except in association with cancer, and it appears to be an early molecular event associated with colorectal cancer monitoring of its expression may be useful as a biomarker for the early detection of this disease