Lectin-like characteristics of recombinant human interleukin-1beta recognizing glycans of the glycosylphosphatidylinositol anchor

We found that 35S-labeled recombinant human interleukin-1beta (rhIL-1beta) binds phosphatidylinositol-specific phospholipase C-treated human placental alkaline phosphatase, phosphatidylinositol-specific phospholipase C-treated trypanosome surface variant glycoproteins, and urinary uromodulin immobilized on plates or immobilized on CNBr-activated Sepharose 4B. The interaction between rhIL-1beta and these glycoproteins was lectin-like, since it was inhibited in the presence of specific saccharides, i.e. mannose 6-phosphate or synthetic Ac-NH.CH2.CH2. PO4--->6Manalpha1-->(+/-2Manalpha1-->+/-6Manalpha1-->) propyl at about 1 microM. On the other hand, a wide variety of compounds including biantennary sugar chains derived from these glycoproteins as well as ethanolamine phosphate, inositol phosphate, mannose 6-sulfate, mannose 1-phosphate, glucose 6-phosphate, and mannitol 6-phosphate did not show any inhibitory effect at concentrations up to 1 mM. These results indicate that rhIL-1beta interacts with these glycoproteins via the mannose 6-phosphate diester of glycans on the glycosylphosphatidylinositol (GPI) anchor. Furthermore, when monolayers of polarized Madin-Darby canine kidney cells on polycarbonate filter membranes were incubated with 35S-rhIL-1beta in either the apical or basolateral chamber, 35S-interleukin-1beta was found to bind specifically to the apical membranes with a Ka value of 4.6 x 10(7) M-1, and the specific interaction was inhibited by 1 microM mannose 6-phosphate. Since the mannose 6-phosphate diester moiety exists only in the GPI glycans on plasma membranes, it was evident that interleukin-1beta can directly interact with the mannose 6-phosphate diester component of the intact glycan of GPI anchors on plasma membranes.     

Distinct promoter sequences of two precursor genes for salmon gonadotropin-releasing hormone in masu salmon

Three phase partitioning (TPP) uses t-butanol and ammonium sulfate to precipitate enzymes and proteins from aqueous solutions. The method is useful both upstream with crude samples and downstream where a scaleable simple step is needed. About 25 enzymes and proteins have been isolated by various laboratories using TPP-t-butanol. The relation of t-butanol used in TPP, with n-butanol used as an extraction agent from Morton's work, is reviewed. Some t-butanol appears bound to TPP-precipitated proteins which are actually protein-t-butanol coprecipitates. They float above denser aqueous salts because bound t-butanol increases their buoyancy, similar to the behavior of many lipoproteins. On redissolving TPP-precipitated enzymes, total and specific activities usually are regained and sometimes increased. Sulfate ion-in large concentrations-likely exerts itself through its kosmotropic action as in conventional salting out. t-Butanol likewise appears to be a kosmotrope and crowding agent at room temperature or above, whereas C1 and C2 cosolvents (e.g., ethanol) do not so behave except at near or below zero temperatures. However, kosmotropy is not the entire origin of TPP, nor probably of conventional salting out. Electrostatic forces, capacity to force protein conformation tightening and protein hydration shifts, also contribute. Electrostatic forces, and the tendency for salt ions to bind and tighten protein molecule conformation, are indicated by the sharp pH dependency of both conventional salting out and TPP, around pH regions where proteins undergo conformation changes. Sulfate anion is densely-perhaps extraordinarily-hydrated, adding much to its effective size, and therefore it has a tendency to crowd or exclude proteins, when sulfate concentrations are in the 0.5 to 3 M range.     

Structural properties and electrical behaviour of thin silicon oxynitride layers

Thin SiO₂ and SiO_xN_y layers were grown on silicon using Rapid Thermal Processing (RTP) in either O₂ or N₂O ambient. Subsequent annealing or nitridation was performed in order to improve the electrical stability. The composition of the films, in particular the incorporation of nitrogen and hydrogen, has been studied. We obtained the distribution of states at the Si/insulator interface through the evaluation of CV measurements and investigated the charge trapping in the layers analysing the voltage–time behaviour during Fowler–Nordheim constant current injection. Furthermore, assuming a trap assisted tunneling mechanism, the influence of near interface trap states on the current voltage characteristic was used to derive an effective insulator state distribution.     

Electrochemically induced asymmetrical etching in InAlAs/InGaAs heterostructures for MODFET gate-groove fabrication

We have achieved a self-controlled asymmetrical etching in metalorganic chemical vapor deposition-grown InAlAs/InGaAs heterostructures, which can be suitable for fabricating modulation-doped field-effect transistors (MODFETs) with gate-groove profiles for improved performance. The technology is based on electrochemical etching phenomena, which can be effectively controlled by using different surface metals for ohmic electrodes. When surface metals of Pt and Ni are deposited on the source and the drain, respectively, the higher electrode potential of Pt results in slower etching on the source side than on the drain side. Thus, asymmetry of gate grooves can be formed by wet-chemical etching with citric-acid-based etchant. This represents a new possibility to conduct “recess engineering” for InAlAs/InGaAs MODFETs.     

Effects of short- and long-term dexamethasone treatment on growth and growth hormone (GH)-releasing hormone (GRH)-GH-insulin-like growth factor-I axis in conscious rats

Although the inhibitory effects of a chronic excess of glucocorticoids (GC) on body growth and GH secretion are well established, the mechanisms involved remain unclear. In this study, we examined the chronic effects of a high dose of dexamethasone (DEX) on spontaneous GH secretion and insulin-like growth factor (IGF)-I in conscious rats. The animals were given daily i.p. injections of DEX (200 microg/day) for either one or four weeks. Body growth assessed by tibia length and serum IGF-I levels was significantly inhibited 1 week after treatment. By contrast, spontaneous GH secretion was not altered 1 week after the treatment. Neither hypothalamic GRH and somtatostain mRNA levels nor GH responses to GRH from single somatotropes were affected 1 week after the treatment. Four weeks after DEX treatment, body growth of the rats was noticeably suppressed. Interestingly, spontaneous GH secretion, hypothalamic GRH mRNA levels and GH responses to GRH were all inhibited 4 weeks after treatment. Pituitary GRH receptor mRNA levels were not altered 1 week after treatment, but increased after 4 weeks. These results indicate that a high dose of DEX initially impairs IGF-I production and subsequently inhibits spontaneous GH secretion in rats. Inhibition of spontaneous GH secretion resulting from chronic GC excess is due, at least in part, to the impairment of hypothalamic GRH synthesis and pituitary GH responsiveness. An increase in the pituitary GRH receptor may be caused by decreased GRH secretion.     

The significance of urinary N-acetyl-beta-D-glucosaminidase for predicting early stage diabetic nephropathy

We studied urinary N-acetyl-beta-D-glucosaminidase (NAG) in the early stage of diabetic nephropathy in 27 non-insulin-dependent diabetes mellitus (NIDDM) patients with a microalbumin level below 20 mg on 24-hour urine sample. Microalbumin and NAG excretion were measured in 24-hour urine samples collected on three separate occasions within seven days of admission. Creatinine clearance was determined simultaneously. There was a significant negative correlation between the creatinine clearance and 24-hour urinary NAG (r = -0.38, p < 0.05). Elevation of urinary NAG may indicate decreased renal function during early stage NIDDM nephropathy.     

Bladder pseudotumor in childhood

The application of frozen and radiation sterilized allogenic bone grafts for reconstructions in orthopaedic operations is described. Analysis of results of treatment of 1125 patients was performed. It was found that use of preserved bone allows to reduce the extend and duration of surgery. Nearly total substitution of grafts may be seen in 3 to 8 months after surgery.     

Subcutaneous soft tissue tumours at the site of implanted microchips in mice

An experiment using 4279 CBA/J mice of two generations was carried out to investigate the influence of parental preconceptual exposure to X-ray radiation or to chemical carcinogens. Microchips were implanted subcutaneously in the dorsolateral back for unique identification of each animal. The animals were kept for lifespan under standard laboratory conditions. In 36 mice a circumscribed neoplasm occurred in the area of the implanted microchip. Females were significantly more frequently affected than male mice. An influence of age or different treatment on the s.c. tumour incidence in two mice generations could not be observed. Macroscopically, firm, pale white nodules up to 25 mm in diameter with the microchip in its center were found. Microscopically, soft tissue tumours such as fibrosarcoma and malignant fibrous histiocytoma were detected.     

Intelligent control of the feeding of aluminum electrolytic cells using neural networks

To be efficient, the control of alumina feeding of the electrolytic cell must be based on cell resistance, alumina concentration, and cell state. Most control schemes now in use are based on cell resistance only, and, thus, constitute an open-loop control that lacks robustness because their decision criteria are not explicitly tied to concentration nor to cell state. This results in the cell operating at nonoptimal concentrations, and cell efficiency is diminished. An optimal operation requires a knowledge of concentration and an adjustment of the decision criteria as a function of concentration. A learning vector quantization (LVQ) type of neural network was built and trained to recognize the cell state. Knowing the state of the cell and its resistance, concentration can be estimated using predetermined regression functions. The decision criteria for the control logic are then consequently adapted. A closed-loop control scheme is thus obtained. Results show that, with its control so structured, the cell can operate at or near optimal concentrations independently of its state. This flexible and intelligent character of the neural control can provide a considerable advantage as compared to the standard control.