Triggering of cellulase biosynthesis by cellulose in Trichoderma reesei. Involvement of a constitutive, sophorose-inducible, glucose-inhibited beta-diglucoside permease

We prepared [U-14C]cellobiose by cultivating Acetobacter pasteurianus in the presence of [U-14C]glucose and hydrolyzing the [U-14C]cellulose formed with beta-glucosidase-free cellulase from Trichoderma reesei. This 14C-labeled cellobiose was used to investigate the presence of an uptake system for cellobiose in T. reesei. Evidence was obtained for the presence of a high affinity (Km for cellobiose 0.3 microM) but low activity (2.5 milliunits/mg fungal dry weight) cellobiose permease. The permease is formed constitutively, but higher levels are formed after addition of sophorose (glucosyl-beta-1,2-diglucoside), a reputed cellulase inducer. The permease appears to be specific for beta-diglucosides, as the uptake of [U-14C]cellobiose is inhibited by sophorose, gentiobiose (glucosyl-beta-1,3-glucoside), and cellobiose. Under these conditions, cellooligodextrines (n, 4-7; final concentration, 1 mM) are not inhibitors. Glucose, but no other monosaccharides, inhibits the permease. The hypersecretory mutant T. reesei RUT C-30 exhibits elevated permease activities, whereas in T. reesei QM 9979, a mutant strain defective in the induction of cellulases by cellulose or sophorose, strongly reduced permease activities were demonstrated. The results stress a hitherto not recognized point of control in the induction of cellulases by T. reesei at the level of uptake of cellulose oligosaccharides.     

Cortisone-induced osteoporosis: effects on bone adenylate cyclase

The majority of patients with soft tissue or bone sarcomas of the upper extremity can be treated today with limb-saving procedures using combined modality therapies. For patients with a tumor in the shoulder area, sometimes an interscapulothoracic amputation is the only radical surgical treatment. However, in selected cases, in which the tumor does not involve the neurovascular bundle, a limb-sparing alternative might be the Tikhoff-Linberg resection. Normal function of the hand and forearm, with reasonable function of the elbow, can be maintained by this procedure. Four case histories are reported.     

Large specimen bone embedment and cement line staining

Undecalcified embedment of large bone specimens is often challenging. A method is presented here that is suitable for methacrylate embedment of sections of canine vertebrae while retaining the ability to localize tartrate-resistant acid phosphatase and alkaline phosphatase activity. Specimens also retained tetracycline labelling, and sectioned preparations were readily stained with routine bone procedures. A modification of the Bodian silver stain, used for examining the nerves and spinal cord in these specimens, provided a useful stain for canaliculi and cement lines in trabecular and cortical bone. This stain is advantageous when both bone and nerve tissue are of interest, as in spinal fusion studies.     

High quality LTCC resonator for voltage-controlled oscillator

Design and technology of microwave conductor lines embedded in low-temperature cofired ceramic (LTCC) multilayer substrates are summarized with a focus on achieving the highest possible quality (Q) factor for a given line inductance. The work was initiated to test the integrability of base station voltage-controlled oscillators (VCOs) in ceramic multilayer substrates. This approach leads to a miniaturization of current versions by a factor of 2 to 4. However, base station specifications for phase noise and hence resonator Q are extremely demanding. Therefore, both the design and the processing technology were optimized. By choosing a twin-line design with two parallel lines vertically separated by a single LTCC layer, Q factors of 90 and 180 have been achieved for integrated 5.5 nH inductors at frequencies of 640 MHz and 1650 MHz, respectively. Application of this result to VCO modules in standard LTCC technology already yields low phase noise levels, e.g., -136 dBc/Hz at 100 kHz offset, which is suitable for base station applications. However, further noise reduction is expected from a dedicated high Q fabrication process that uses conventional via punching and filling steps to replace the ceramic material between the two lines by conductive silver paste. This raises the Q to 120 and 200, respectively, at the two frequencies and adds extra degrees of freedom to LTCC design for low-loss wireless solutions.     

External surface and lamellarity of lipid vesicles: a practice-oriented set of assay methods

Three methods are presented for the determination of external surface of large lipid vesicles of different lamellarity with 2% absolute accuracy. These methods (referred to as EPR, NBD and TNBS assays) use different marker lipids which provide signals (electron paramagnetic resonance, fluorescence of N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl) residues, and UV absorption increase of 2,4,6-trinitrobenzenesulfonic acid after reaction with aminolipids, respectively). The signals change upon addition of different membrane-impermeant reagents due to reaction with marker lipids at the external vesicle surface. They were applied to the same vesicle samples, including unilamellar and multilamellar vesicles, both at two different lipid compositions. External surface data matched for the three assays within 2%, but only after appropriate redesign or adaptation of so far published procedures. Main improvements related to slow influx of reagents (TNBS and NBD assays) or to redistribution of marker lipids (EPR assay), obscuring determination of outer vesicle surface from fast reaction between reagent and readily accessible marker lipids. Furthermore, suitable strategies were found to obtain accurate 100% values (reaction of all marker lipids present), required to relate external vesicle surface to total surface. This included corrections for light scattering (NBD assay) and for turbidity (TNBS assay). These three methods appear to close a gap in the methodology to determine external surface of vesicles for typical practical needs. In particular, the reliability range of the NBD assay could be extended to marker lipid densities as low as 1 marker lipid per 3000 lipids.     

Tree-based partitioning of date for association rule mining

The most computationally demanding aspect of Association Rule Mining is the identification and counting of support of the frequent sets of items that occur together sufficiently often to be the basis of potentially interesting rules. The task increases in difficulty with the scale of the data and also with its density. The greatest challenge is posed by data that is too large to be contained in primary memory, especially when high data density and/or low support thresholds give rise to very large numbers of candidates that must be counted. In this paper, we consider strategies for partitioning the data to deal effectively with such cases. We describe a partitioning approach which organises the data into tree structures that can be processed independently. We present experimental results that show the method scales well for increasing dimensions of data and performs significantly better than alternatives, especially when dealing with dense data and low support thresholds. Shakil Ahmed received a first class BSc (Hons) degree from Dhaka University, Bangladesh, in 1990; and an MSc (first class), also Dhaka University, in 1992. He received his PhD from The University of Liverpool, UK, in 2005. From 2000 onwards he is a member of the Data Mining Group at the Department of Computer Science of the University of Liverpool, UK. His research interests include data mining, Association Rule Mining and pattern recognition. Frans Coenen has been working in the field of Data Mining for many years and has written widely on the subject. He received his PhD from Liverpool Polytechnic in 1989, after which he took up a post as a RA within the Department of Computer Science at the University of Liverpool. In 1997, he took up a lecturing post within the same department. His current Data Mining research interests include Association rule Mining, Classification algorithms and text mining. He is on the programme committee for ICDM'05 and was the chair for the UK KDD symposium (UKKDD'05). Paul Leng is professor of e-Learning at the University of Liverpool and director of the e-Learning Unit, which is responsible for overseeing the University's online degree programmes, leading to degrees of MSc in IT and MBA. Along with e-Learning, his main research interests are in Data Mining, especially in methods of discovering Association Rules. In collaboration with Frans Coenen, he has developed efficient new algorithms for finding frequent sets and is exploring applications in text mining and classification.     

Antisense Oligonucleotide-Based Rescue of Aberrant Splicing Defects Caused by 15 Pathogenic Variants in ABCA4

The discovery of novel intronic variants in the ABCA4 locus has contributed significantly to solving the missing heritability in Stargardt disease (STGD1). The increasing number of variants affecting pre-mRNA splicing makes ABCA4 a suitable candidate for antisense oligonucleotide (AON)-based splicing modulation therapies. In this study, AON-based splicing modulation was assessed for 15 recently described intronic variants (three near-exon and 12 deep-intronic variants). In total, 26 AONs were designed and tested in vitro using a midigene-based splice system. Overall, partial or complete splicing correction was observed for two variants causing exon elongation and all variants causing pseudoexon inclusion. Together, our results confirm the high potential of AONs for the development of future RNA therapies to correct splicing defects causing STGD1.     

Evaluation and characterization of ammoniumpolyphosphate–pentaerythritol‐based systems for intumescent coatings

Intumescent coatings are increasingly used as a method of passive fire protection on steel constructions. By forming a carbon network and releasing a blowing agent, the thin intumescent film swells 100‐fold at elevated temperatures. The highly insulating foam effectively prevents the load bearing steel from reaching its critical temperature at which it looses its mechanical properties and collapses. The role of the carbon donor in intumescent coatings has been studied. Comparison in temperature development, foaming ratios, and rheological behavior has been performed between formulations containing pentaerythritol (penta), di–penta, and tri–penta. A simulated fire test, in which the temperature development during intumescence was studied, showed that the formulations containing penta were considerably more efficient in keeping a low temperature throughout the process. A more rapid temperature development was displayed when using di–penta and tri–penta as the carbon donor. Rheometer tests indicate that penta formulations enter the intumescent process at a lower temperature and stays in it for a longer time than di–penta and tri–penta formulations. Furthermore, the crossover temperature and maximum phase angle are shifted towards higher temperatures by replacing penta with di–penta and with tri–penta in the formulations, respectively. © 2007 Wiley Periodicals, Inc. J Appl Polym Sci 104: 748–753, 2007