Regeneration of active receptor recognition domains on the B subunit of cholera toxin by formation of hybrids from chemically inactivated derivatives

In order to test the hypothesis that binding sites of cholera toxin for its receptor, the monosialoganglioside GM1, are shared between adjacent beta-polypeptide chains, two inactive chemical derivatives of the B subunit of cholera toxin (CTB) were prepared and were subsequently used for the construction of hybrid CTB pentamers. One inactive derivative consisted of CTB specifically modified in the single essential Trp-88 residue of each beta-chain. This residue was modified by formylation, a treatment preserving the structural integrity of CTB. The other inactive derivative consisted of CTB specifically succinylated in three amino groups located in or near the receptor binding site. Using [1,4-14C]succinic anhydride for the site-specific succinylation and analysis of radiolabeled tryptic fragments of S-carboxymethylated [14C]sssCTB revealed that the amino groups specifically modified were the alpha-amino group of Thr-1 and the epsilon-amino groups of respectively Lys-34 and Lys-91. Upon submitting equal amounts of formylated CTB and site-specific succinylated CTB to a denaturation-renaturation cycle, hybrid pentamers were formed which in contrast to the parental compounds were able to bind GM1. The affinity of hybrid CTB for GM1, as estimated by a competitive solid-phase radiobinding assay was unexpectedly high and only 2.5-fold lower than that of its native counterpart. The number of active binding sites on hybrid CTB was determined from: (i) titration with the oligosaccharide moiety of GM1 (oligo-GM1) and monitoring the reversal of the Trp fluorescence quenching by iodide ions and (ii) rapid gel filtration over a superdex HR column of a mixture of hybrid CTB and an excess of 3H-labeled oligo-GM1. The data are in agreement with the formation of one active binding per four reconstituted binding sites in hybrid CTB, which is consistent with a random association of CTB monomers during the denaturation-renaturation cycle.     

Practical accuracy analysis of some existing effective channellength and series resistance extraction methods for MOSFET's

A method to analyze the accuracy of the extracted values for the channel length (L_eff) and series resistance (R_s) of MOSFET devices is presented. The analysis is based on a statistical argument being the variance σ of the extracted results. This variance is found to be a good measure for the accuracy of the particular extraction method used. It is shown that, in the case of deep submicron technologies, errors as large as 200 nm for ΔL can be made for these extraction methods depending on the process design and the process control. The use of a single transistor method is suggested as a possible solution to the low accuracy of the L-array methods     

Plasma mevalonate concentrations in uremic patients

Mevalonic acid (mevalonate or MVA), is an obligate precursor in the biosynthetic pathway of cholesterol. It is partially metabolized by the kidneys and its plasma concentrations are an index of endogenous cholesterol synthesis. The aim of the present study was to evaluate plasma MVA concentrations in uremic patients with different degrees of chronic renal failure (CRF; group A), and the effects of a single hemodialysis treatment on plasma MVA in a group of patients with end-stage renal disease (ESRD; group B). CRF patients exhibited a higher mean basal mevalonate concentration (13.3 +/- 6.5 ng/ml) than control subjects (4.68 +/- 1.32 ng/ml; P < 0.001). A statistically significant direct correlation was evident in CRF patients between mevalonate and creatinine plasma levels (r = 0.86; P < 0.001). A single hemodialysis treatment was associated with a significant reduction of plasma mevalonate concentrations four hours after the hemodialysis session (-57%; P < 0.001) and an increase up to the basal values 24 hours after the end of the treatment. In conclusion, our results demonstrated: (i) higher plasma MVA concentrations in patients with decreased renal function; (ii) a direct relationship between plasma MVA levels and the degree of kidney failure as expressed by creatinine plasma concentrations; and (iii) a clear cut reduction of elevated plasma MVA levels after a single hemodialysis treatment.     

Stability, in-Vitro and in-Vivo Release Studies from Metronidazole Ointments

The influence of ointment formulation on the stability, the in-vitro release and the in-vivo absorption through the skin of rabbits was investigated. The choice of the selected ointments has no influence on the drug stability with the exception of an acidified emulsion base. A good correlation between in-vitro release and in-vivo absorption was found revealing that metronidazole was quickly released and effectively absorbed from a polyethylene glycol base.     

Specific bronchial provocation tests with flour in the diagnosis of occupational bronchial asthma

The gold standard in the diagnosis of occupational asthma is the specific bronchial provocation test (sBPT), but other diagnostic criteria have been proven to have a similar sensitivity, mainly in asthma due to high molecular weight compounds. In order to assess wether some clinical findings can predict the positive response to sBPT, we studied 37 subjects (14 millers and 23 bakers) with suspected occupational asthma who underwent sBPT with wheat flour dust (dust exposure in a small cabin: geometric mean 12.1 mg/m3 for up to 30 min). A positive response to sBPT (FEV1 > 20%) was elicited in 20 subjects (11 early, 4 late, and 5 dual responses). There was no significant difference between subjects with positive or negative sBPT as regards mean age, smoking, length of employment, duration of symptoms, atopy (skin positivity to one or more common allergens) and PD20FEV1 methacholine. The percentage of subjects with work-related symptoms was significantly higher in subjects with positive sBPT with respect to subjects with negative sBPT (81% versus 41.2%, p < 0.01 by chi 2 test); furthermore, FEV1 was significantly lower in subjects with positive sBPT. The percentage of positive skin response to wheat flour extract (mean wheal diameter > or = 3 mm) was mildly but not significantly higher in subjects with positive sBPT (68.4% versus 41.2%). None of the following clinical factors (age < 35 years, asthma symptoms pre-existing occupational exposure, non smokers, atopy and bronchial hyperresponsiveness to methacholine), alone or in combination, were associated with higher prevalence of positive sBPT. We conclude that the response to sBPT in subjects with suspected occupational asthma due to flour dust can not be adequately predicted by other clinical, allergologic and functional data. Therefore, sBPT with flour dust should always be performed in subjects with suspected occupational asthma.     

Structural and surface properties of thermally prepared Ti/RhxTi(1−x)Oy electrodes: I—influence of preparation parameters

Electrodes of Rh_xTi_(1−x)O_y nominal composition were prepared by thermal decomposition of the chloride or nitrate precursor salts dissolved in strongly acidic medium and applied by brush to both sides of a Ti° support. A systematic study of the influence of calcination temperature and time as well as oxygen flux was conducted. The coatings were characterised by SEM, EDAX, XRD, open circuit potential measurements and cyclic voltammetry (CV). Visible-ultraviolet spectrophotometry was employed to identify the chemical form of the precursor in solution while thermogravimetric analysis (TGA) was used to assess the decomposition temperature ranges. Optimisation of the coating preparation parameters showed coatings obtained from [Rh(H₂O)₆](NO₃)₃ precursor dissolved in HNO₃ 1:2 (v/v) and fired at 430 °C for 2 h in a 5 l min⁻¹ oxygen stream-furnished stable electrodes having the highest electrochemically active surface area.     

Room temperature synthesis of crystalline metal oxides

Crystalline titanium dioxide powders have been synthesized as either rutile or anatase from aqueous solutions at low temperatures (T≤100°C) and atmospheric pressure. First, a sol is prepared by the hydrolysis of a titanium alkoxide in an acidic solution. The sol is subsequently heated at different rates to produce the different crystalline phases of titanium dioxide. Powder characterization was carried out using X-ray diffraction, scanning electron microscopy and high resolution transmission electron microscopy. In general, the precipitate size was observed to be between 50 and 100 nm. Possible mechanisms involved in determining the crystal variants are discussed. This revised version was published online in November 2006 with corrections to the Cover Date.     

Photoelectronic and transport properties of polypyrrole doped with a dianionic dye

Polypyrrole (PPy) doped with dodecylsulfate (DS) and an organic dye (indigo carmine, IC) was electrochemically prepared and characterized by Raman spectroscopy and X-ray diffraction (XRD). The photoelectrochemical properties of PPy-DS and PPy-DS-IC in contact with an electrolytic solution containing a redox couple were studied using the theories for the semiconductor ∣ electrolyte interface. Results indicate that the system containing IC presents a higher photocurrent density under polychromatic illumination and faster response time when compared with PPy-DS. This fact was assigned to the formation of molecular-scale paths in PPy-DS-IC. The IC molecules lying perpendicularly between PPy chains, as confirmed by the X-ray analysis, could facilitate the mass transport at the interface and increase the ordering degree to provide better electronic charge transfer in the bulk. Although the photoelectrochemical devices presented here do not present all the properties of inorganic-based devices, we discuss some strategies to enhance the photoelectrochemical properties and response time of conducting polymers used in these type of systems.     

Studies on the electrical conductivity and galvanomagnetic characteristics of short carbon fibre filled thermoplastic elastomers

The electrical resistivity of short carbon fibre filled thermoplastic elastomers based on natural rubber-high density polyethylene (NR-HDPE) blend and styrene-isoprene-styrene (S-I-S) block copolymer has been studied by varying the concentration of fibre from the 0 to 40 phr level. A sharp drop in resistivity is observed in both cases after a critical concentration of fibre is reached, with the S-I-S system showing lower values of critical concentration. At higher concentrations, experimental values of the electrical conductivity agreed reasonably well with the calculated values from a theory based on the probability of formation of a conductive network. The effect of temperature on the resistivity of the composites has also been investigated in a temperature range of 27–120 °C and 27–90 °C for the NR-HDPE system and S-I-S system, respectively. The type of carrier in both the composites is found to be n-type, i.e. electrons, and the activation energy of hopping of the electrons for both the systems has been calculated. A Hall effect study has also been carried out to determine the carrier concentration and drift mobility of both the composites.     

Preparation of V/ZrO2 catalysts by the sol-gel method: Physical and structural characterization

Some zirconia-supported vanadium oxides have been prepared by sol-gel method and characterized by different techniques. The vanadium content exerts an effect on the area and pore-size of precursors and samples calcined. The presence of vanadium also affects the decomposition of precursors into ZrO₂ and to its phase transformation, as well as the formation of fibres, as shown by SEM-EDX. XPS analyses show the presence of two species of vanadium at the surface, V^5– and V⁴⁺, the percentage of each one depending on the load of vanadium in the catalyst.