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.     

Human neonatal ovary: proposal of a three-dimensional model

A model of human neonatal ovary is presented, derived from morphometric, evaluations carried out on left ovaries removed from five full-term neonates with a 46, XX karyotype, free from malformations of the genital apparatus. According to this model, the gonad can be represented by a triaxial ellipsoid with a central medullary core surrounded by a cortical stratum of constant thickness. The germinal population, consisting of follicles and primitive cortical tissue, occupies the cortex, intermingled with the interstitium or stroma. In the cortex it is then possible to describe an outer layer formed by primitive cortical tissue, and an inner portion occupied by follicles. The primary and secondary follicles fill the portion near the medulla and the primordial ones are contained in the middle and outer zones. Since the variability observed among ovaries is slight, we can propose a mean model of neonatal ovary in which the spatial relationships among the different components, the total number of follicles and their position in the cortex can be calculated.     

Effect of the approximately 2400-year solar climatic cycle on the lives of peoples

2400-year solar cycle ascertained by us in the long-term series of various natural characteristics (e.g. the radiocarbon content in samples of well-known ages, oscillations of the sea and ocean levels etc.) includes three parts roughly equal in duration: a phase of high activity, a phase of depression and a phase of moderate activity. In climate oscillations, which are connected with this cycle, the next phenomena are established: a little climatic optimum, a little Ice Age, and a temperate climate epoch.     

Clinical and historical predictors of dental caries on radiographs

Jet injection can be used to introduce genes into the cells of differentiated tissues of living animals and organ cultures. When a solution of plasmid DNA is jet injected into a selected tissue or organ, cells lying in or near the path of the jet injection are transfected with the DNA and the introduced gene(s) are expressed. Since there is minimal morbidity from each jet injection, multiple injections can be performed at the same or nearby sites. Both mRNA and protein expression from transfected genes can be quantitated using standard methods. In addition, the technique is an efficient means of DNA immunization. Methodology for using jet injection to transfer plasmid DNA into the cells of skin, fat, mammary gland, and muscle are described.     

The efficacy of Augmentin in suppurative complications in neurosurgery]

The results of clinical and laboratory studies on the use of augmentin in severe purulent complications after neurosurgical operations are presented. The laboratory studies carried out with the use of an automatic system Cobas Bact (Roch) showed that the numbers of the augmentin resistant strains of Staphylococcus and Enterobacteriaceae among the pathogens were 47 and an average of 64.5%, respectively. Gram-negative bacteria resistant to augmentin were 1.5 to 2 times less frequent than those resistant to amoxycillin. Still, they were much more frequent than those resistant to cefotaxime and ceftriaxone. Clinical efficacy of augmentin was studied in treatment of 39 patients with various affections of the brain such as tumors, trauma, vascular malformations and inflammatory processes. The postoperative complications were represented by meningitis, pneumonia, sepsis and their associations. The use of augmentin in the severe intra- and extracranial complications was favourable in 82.1% of the cases.     

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.