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.     

Synthesis and characterization of molybdenum disulphide formed from ammonium tetrathiomolybdate

An investigation has been carried out into the possibility of in situ formation of MoS2 within porous anodic films on aluminium, to improve subsequent tribological behaviour, by re-anodizing in thiomolybdate electrolyte. Acidification of thiomolybdate was employed to simulate the conditions for formation of the sulphide at the anodic film/electrolyte interface, followed by appropriate vacuum heat treatments to study possible temperature effects on the sulphide due to either friction or Joule heating during anodizing. The products of both acidification and heat treatment, characterized by X-ray powder diffraction and scanning electron microscopy, were compared with those formed by direct thermal decomposition of ammonium tetrathiomolybdate crystals. The precipitate formed by acidification was mainly amorphous molybdenum trisulphide (MoS3), which on heat treatment at 450 and 850°C yielded 3R-MoS2. 3R-MoS2 also formed by the thermal decomposition of thiomolybdate crystals. Thermogravimetric and differential thermal analyses showed that the decomposition of MoS3 to MoS2 occurred in the range 220–370°C and revealed the sequence of reaction steps. The findings suggest that mainly amorphous MoS3 is formed as a consequence of changes in the pH of the film/electrolyte interface during re-anodizing but the product is relatively easily transformed to crystalline MoS2 on moderate heating which may occur during wear processes.     

Identification of the DRB1*1313 allele in a sibling pair

Amines such as dopamine, norepinephrine and epinephrine were analysed in the brain regions of O.mossambicus exposed to quinalphos, phenthoate and their combination for 96 hr. The three types of treatments significantly (P < 0.05) altered the amines level at various intervals in the brain regions.     

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.     

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.     

Late rectal sequelae following definitive radiation therapy for carcinoma of the uterine cervix: a dosimetric analysis

OBJECTIVE: The structure of the collagen scar during healing of a myocardial infarction is a determinant of the function of the remodeled tissue. We hypothesize that the passive deformations of both scar and normal tissue are related to the underlying collagen uncoiling as the tissue stretches, and that the unloaded tortuosity of the collagen may be a determinant of tissue stiffness at low ventricular pressure. Hence collagen uncoiling and tissue strain were measured during passive loading in normal tissue, and in healing infarct tissue. METHODS: Left ventricles of rats were infarcted by ligation of the left anterior descending artery for 2 weeks. Surface strains were measured during passive inflation in the scar region in one set of excised hearts, and other arrested hearts were fixed at different ventricular pressures, after which collagen tortuosity was measured in the infarcted and normal tissue. RESULTS: Passive loading strains were smaller in the scar in both the fiber and cross-fiber directions. Tortuosity decreased with load in normal and infarcted tissue, with fibrils tending to straighten more in the scar tissue at higher pressures (1.056 +/- 0.009 vs. 1.024 +/- 0.009 at P = 20 mmHg) with similar tortuosities at zero pressure (1.110 +/- 0.012 vs. 1.098 +/- 0.019). The decrease in tortuosity with strain was greater for the infarcted tissue. CONCLUSIONS: The greater stiffness of infarcted tissue at low pressure is not due to 'straightened' collagen fibers, and there may be a different three-dimensional structure of infarct vs. normal coiled collagen fibers which can affect the material properties of these tissues.