Detection and quantification of neurotensin in human brain tissue by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry

A method was developed for mass spectrometric detection of neurotensin (NT)-like immunoreactivity and quantification of NT in human brain tissue. The method is based on immunoprecipitation followed by analysis using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). The identity of the major component of the immunoprecipitates as neurotensin was confirmed by fragment ion analysis on an electrospray ionization quadrupole time-of-flight instrument. MALDI-TOF-MS quantification of NT was achieved using stable-isotope-labeled NT as the internal standard, yielding an error of less than 5%. The method allowed detection of low-femtomole amounts of NT, staring from low-milligram amounts of lyophilized brain tissue. In addition to NT, several other peptides were detected in the purified samples, most of which, according to their molecular masses, corresponded to fragments of NT. The method is demonstrated with quantification of NT from human hypothalamus tissue, and a comparison is made with results obtained from competitive radioimmunoassay.     

Multipotentiality among the intellectually gifted: "It was never there and already it's vanishing."

The theory of work adjustment was used as a conceptual framework in evaluating the concept of multipotentiality, taken from the psychological literature on counseling intellectually gifted individuals (viz., those with high-flat ability and preference profiles that may lead to career indecision and distress). An examination of over 1,000 intellectually gifted students (top 1%) in 4 separate cohorts, assessed with the Scholastic Aptitude Test, the Study of Values, and J. L. Holland's (1985) six interest themes, revealed little empirical support for the prevalence of multipotentiality within intellectually talented adolescents (     

Rediscovery of Cerebrosterol

24S-hydroxycholesterol was identified more than half a century ago and was initially given the name "cerebrosterol" due to the fact that it was abundant in the brain. A decade ago, we showed that the most important mechanism by which cholesterol is eliminated from the mammalian brain involves a hydroxylation into cerebrosterol followed by diffusion of this steroid over the blood-brain barrier. Using an (18)O(2) inhalation technique, we showed that about two-thirds of the cholesterol synthesis in rat brain is balanced by conversion into cerebrosterol. The hydroxylase responsible for the reaction was found to be dependent upon NADPH and oxygen, consistent with involvement of a species of cytochrome, P-450. The gene coding for the cytochrome P-450 responsible for the reaction was later cloned by the group of David Russell in Dallas and the enzyme was found to be located to neuronal cells in the brain. Recent studies by us and others on this new pathway for elimination of cholesterol from the brain have given new insights into the mechanisms by which cholesterol homeostasis is maintained in this organ. In addition, these studies have resulted in new diagnostic and prognostic tools in connection with neurological and neurodegenerative diseases. An overview of the studies is presented here and the possibility is discussed that the cholesterol 24S-hydroxylase in the brain may be a new drug target in connection with neurodegenerative diseases.     

Universally applicable model for the quantitative determination of lake sediment composition using fourier transform infrared spectroscopy

Fourier transform infrared spectroscopy (FTIRS) can provide detailed information on organic and minerogenic constituents of sediment records. Based on a large number of sediment samples of varying age (0-340,000 yrs) and from very diverse lake settings in Antarctica, Argentina, Canada, Macedonia/Albania, Siberia, and Sweden, we have developed universally applicable calibration models for the quantitative determination of biogenic silica (BSi; n = 816), total inorganic carbon (TIC; n = 879), and total organic carbon (TOC; n = 3164) using FTIRS. These models are based on the differential absorbance of infrared radiation at specific wavelengths with varying concentrations of individual parameters, due to molecular vibrations associated with each parameter. The calibration models have low prediction errors and the predicted values are highly correlated with conventionally measured values (R = 0.94-0.99). Robustness tests indicate the accuracy of the newly developed FTIRS calibration models is similar to that of conventional geochemical analyses. Consequently FTIRS offers a useful and rapid alternative to conventional analyses for the quantitative determination of BSi, TIC, and TOC. The rapidity, cost-effectiveness, and small sample size required enables FTIRS determination of geochemical properties to be undertaken at higher resolutions than would otherwise be possible with the same resource allocation, thus providing crucial sedimentological information for climatic and environmental reconstructions.     

Piezo dispensed microarray of multivalent chelating thiols for dissecting complex protein-protein interactions

The fabrication of a novel biochip, designed for dissection of multiprotein complex formation, is reported. An array of metal chelators has been produced by piezo dispensing of a bis-nitrilotriacetic acid (bis-NTA) thiol on evaporated gold thin films, prestructured with a microcontact printed grid of eicosanethiols. The bis-NTA thiol is mixed in various proportions with an inert, tri(ethylene glycol) hexadecane thiol, and the thickness and morphological homogeneity of the dispensed layers are characterized by imaging ellipsometry before and after back-filling with the same inert thiol and subsequent rinsing. It is found that the dispensed areas display a monotonic increase in thickness with increasing molar fraction of bis-NTA in the dispensing solution, and they are consistently a few Angstr?ms thicker than those prepared at the same molar fraction by solution self-assembly under equilibrium-like conditions. The bulkiness of the bis-NTA tail group and the short period of time available for chemisorption and in-plane organization of the dispensed thiols are most likely responsible for the observed difference in thickness. Moreover, the functional properties of this biochip are demonstrated by studying multiple protein-protein interactions using imaging surface plasmon resonance. The subunits of the type I interferon receptor are immobilized as a composition array determined by the surface concentration of bis-NTA in the array elements. Ligand dissociation kinetics depends on the receptor surface concentration, which is ascribed to the formation of a ternary complex by simultaneous interaction of the ligand with the two receptor subunits. Thus, multiplexed monitoring of binding phenomena at various compositions (receptor densities) offers a powerful tool to dissect protein-protein interactions.     

Hydrogen-induced CO2 formation from ethylene deposits on Pt during consecutive O2 and H2 exposures

The oxidation of C₂H₄ deposits on polycrystalline Pt when exposed to consecutive O₂ and H₂ pulses at room temperature has been investigated in a long (L = 36 mm), shallow (d = 600–700 nm) micromachined glass–SiO₂–Pt channel. Hydrogen-induced CO₂ formation from species accumulated on the Pt surface was observed. Frequent switching of the O₂/H₂ exposure pulses was found to increase the efficiency of the oxidation of the carbonaceous deposits markedly. The observations may be of general interest for the regeneration of contaminated catalysts.     

Thermal performance of combined solar and pellet heating systems

Various pellet heating systems are marketed in Sweden, some of them in combination with a solar heating system. Several types of pellet heating units are available and can be used for a combined system. This article compares four typical combined solar and pellet heating systems. System 1 and 2 with a pellet stove, system 3 with a store integrated pellet burner and system 4 with a pellet boiler. The often lower efficiency of pellet heaters compared to oil or gas heaters increases the final energy demand. Consequently, heat losses of the various systems have been studied. The systems have been modeled in TRNSYS and simulated with parameters identified from measurements. For almost all systems the flue gas losses are the main heat losses except for system 3 where store heat losses prevail. Relevant are also the heat losses of the burner and the boiler to the ambient. Significant leakage losses are noticed for system 3 and 4. For buildings with an open internal design system 1 is the most efficient solution. Other buildings should preferably apply system 2 or 3. The right choice of the system depends also on whether the heater is placed inside or outside of the heated area. Unlike the expectations and results from other studies, the operation of the pellet heaters with modulating combustion power is not necessarily improving the performance. A large potential for system optimisation exists for all studied systems, which when applied could alter the relative merits of the different system types.     

Influence of microflora on texture and contents of amino acids, organic acids, and volatiles in semi-hard cheese made with DL-starter and propionibacteria

The microflora of semi-hard cheese made with DL-starter and propionic acid bacteria (PAB) is quite complex, and we investigated the influence of its variation on texture and contents of organic acids, free amino acids, and volatile compounds. Variation in the microflora within the normal range for the cheese variety Grevé was obtained by using a PAB culture in combination with different DL-starters and making the cheeses at 2 dairy plants with different time and temperature profiles during ripening. Propionic acid bacteria dominated the microflora during ripening after a warm room period at levels of log 8 to log 9 cfu/g, which was about 1 log unit higher than the total number of starter bacteria and about 2 log units higher than the number of nonstarter lactic acid bacteria. Eye formation was observed during the warm room period and further ripening (at 8 to 10°C). The amounts of acetate, propionate, total content of free amino acids, 2-propanol, and ethyl propionate in the ripened cheeses were related to the number of PAB. A decrease in the relative content of Asp and Lys and increase of Phe over the ripening time were different from what is observed in semi-hard cheese without PAB. The occurrence of cracks was higher in cheeses with more hydrolyzed α_S1- and β-casein, higher content of free amino acids, lower strain at fracture (shorter texture), and a greater number of PAB.