Circulating IgSF Proteins Inhibit Adhesion of Antibody Targeted Microspheres to Endothelial Inflammatory Ligands

Proposed methods for detecting circulatory system disease include targeting ultrasound contrast agents to inflammatory markers on vascular endothelial cells. For antibody-based therapies, soluble forms of the targeted adhesion proteins of the immunoglobulin superfamily (IgSF) reduce adhesion yet were left unaccounted in prior reports. Microspheres labeled simply with a maximum level of antibodies can reduce the diagnostic sensitivity by adhering to proteins expressed normally at a low level, while sparsely coated particles may be rendered ineffective by circulating soluble forms of the targeted proteins. A new microdevice technique is applied to simultaneously measure the adhesion profile to a series of IgSF-protein-coated surfaces. In this investigation, we quantify the in vitro binding characteristics of 5-μm microspheres to oriented intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1) protein-coated surfaces in the presence of human serum at physiological concentrations. Defined regions of a slide were coated with recombinant chimeric Fc-human ICAM-1 and VCAM-1 in variable ratios but constant total concentration. Monoclonal human anti-ICAM-1 or anti-VCAM-1 antibodies in competition with non-binding mouse anti-rabbit antibodies coat the microsphere surface at a constant surface density with variable yet controlled surface activities. Using multiple slide surface IgSF protein and microsphere antibody concentrations, an adhesion profile was developed for the microspheres with and without IgSF proteins from human serum, which demonstrated that exposure to serum reduced microsphere binding, on average, more than 50% compared to the no-serum condition.. The serum effects were limited to antibodies on the microsphere, since binding inhibition was reversed after rinsing serum from the system and fresh antibody-coated microspheres were introduced. This analysis quantifies the binding effects of soluble IgSF proteins from human serum on antibody-based targeted ultrasound detection and drug delivery methods.     

Synthesis,crystal structure,and photoluminescence of zinc complex [Zn(Qina)<Subscript>2</Subscript>(DMSO)<Subscript>2</Subscript>] · 2DMSO

A novel complex [Zn(Qina)₂(DMSO)₂] · 2DMSO (I) (Qina = quinaldic acid, DMSO = dimethyl sulfoxide) has been synthesized and characterized by X-ray diffraction, elementary analysis, and IR spectrum. The unit cell parameters for complex I: a = 8.2884(11), b = 22.015(3), c = 9.0685(12) Å, β = 100.61(0)°, V = 1626.41(40) ų, Z = 2, space group, P2₁/n. In the complex I, the Zn(II) atom was six-coordinated to form a distorted octahedral geometry by two Qina ligands and two dimethyl sulfoxide molecules. The crystal structure is stabilized by hydrogen bonds, C - H... π, π-π stacking interaction between adjacent complexes. The binding of the complex I with calf thymus DNA has been investigated by fluorescence spectroscopic studies. The result suggested that the complex intercalates into DNA base pairs.     

Enantioselective <Emphasis Type="Italic">O</Emphasis>-allylic alkylation of Morita-Baylis-Hillman carbonates with oxime

The enantioselective O-allylic alkylation of acetophenone oxime with various Morita-Baylis-Hillman (MBH) carbonates has been accomplished by the catalysis of a commercially available cinchona alkaloid (DHQD)2PHAL. The corresponding O-allylic products were obtained in moderate to excellent yields up to 96% ee.     

A Novel endo-1,4-β-Mannanase from <Emphasis Type="Italic">Bispora antennata</Emphasis> with Good Adaptation and Stability over a Broad pH Range

An endo-β-1,4-mannanase encoding gene, man5, was cloned from Bispora antennata CBS 126.38, which was isolated from a beech stump. The cDNA of man5 consists of 1,299 base pairs and encodes a 432-amino-acid protein with a theoretical molecular mass of 46.6 kDa. Deduced MAN5 exhibited the highest amino acid sequence identity of 58% to a β-mannanase of glycoside hydrolase family 5 from Aspergillus aculeatus. Recombinant MAN5 was expressed in Pichia pastoris and purified to electrophoretic homogeneity. The specific activity of the final preparation towards locust bean gum was 289 U mg⁻¹. MAN5 showed optimal activity at pH 6.0 and 70 °C and had good adaptation and stability over a broad range of pH values. The enzyme showed more than 60% of peak activity at pH 3.0–8.0 and retained more than 80% of activity after incubation at 37 °C for 1 h in both acid and alkaline conditions (pH 4.0–11.0). The K _m and V _max values were 1.33 mg ml⁻¹ and 444 μmol min⁻¹ mg⁻¹ and 1.17 mg ml⁻¹ and 196 μmol min⁻¹ mg⁻¹ for locust bean gum and konjac flour, respectively. Of all tested metal ions and chemical reagents, Co²⁺, Ni²⁺, and β-mercaptoethanol enhanced the enzyme activity at 1 mM, whereas other chemicals had no effect on or partially inhibited the enzyme activity. MAN5 was highly resistant to acidic and neutral proteases (trypsin, α-chymotrypsin, collagenase, subtilisin A, and proteinase K). By virtue of the favorable properties of MAN5, it is possible to apply this enzyme in the paper and food industries.     

Theoretical investigation of the Anti-Parkinson drug rasagiline and its salts: conformations and infrared spectra

The conformational analysis of rasagiline [N-propargyl-1(R)-aminoindan] was performed by the density functional theory (DFT) B3LYP method using the 6–31++G (d,p) basis set. A single point energy calculations based on the B3LYP optimized geometries were also performed at MP2/6-31++G (d, p) level. The vibrational frequencies of the most stable conformer of rasagiline was calculated at the B3LYP level and vibrational assignments were made for normal modes on the basis of scaled quantum mechanical force field (SQM) method. The influence of mesylate and ethanedisulfonate salts on the geometry of rasagiline free base and its normal modes are also discussed.      

Limitations of the scalp-hair biologic monitor in assessing selenium status in epidemiological investigations

Scalp hair is routinely used to assess exposure to toxic trace elements and nutritional status of some required trace elements. The advantages and disadvantages of hair as a biologic monitor have been comprehensively discussed in the literature for many years. Among the concerns is distinguishing between exogenous and endogenous contributions. Nested in this issue is the longitudinal distribution of a trace element along the hair strand. The typical observation for many elements of interest is that the element concentration increases from the root end to the distal end; and this is attributed to continuing contamination from exogenous sources. In this study we used neutron activation analysis to measure 14 trace elements in 6 mm segments of full-length scalp hair from three healthy members of the same household having light-urban environmental exposure. To extend the data set for selenium, we included three adult female subjects with longer than average scalp hair. From these trace-element concentrations we calculated the root-to-distal end ratios as a profile diagnostic of trace-element distributions. Ratios fall into three diagnostic categories, >1, ≈1, and <1 corresponding to profiles having decreasing root-to-distal concentrations, unchanging concentrations, and increasing concentrations, respectively. Of the 14 elements measured, only Se has R > 1, Zn and S have R ≈ 1, and the remaining 11 elements all have R < 1 in the order: As > I > Hg ≈ Au ≈ Mg ≈ Mn ≈ Sb ≈ Ca > Cu > Al ≈ Ag. R _Se is greater than 1 and increases with hair length (P = 0.02) corresponding to a continuous longitudinal loss of Se in stark and puzzling contrast to the other elements measured. An analogous loss of Se in the nail monitor was not observed leading us to conclude that the nail is less prone to misclassification of selenium status in epidemiological studies.     

Ferrocenyl-substituted α,β-unsaturated ketones in synthesis of tetrahydropyrimidinones

Ferrocenyltetrahydropyrimidin-2-ones were prepared by reactions of linear and cyclic ,-unsaturated ketones of the ferrocene series with urea in i-PrOH in the presence of t-BuOK. The structures of the compounds prepared were studied by ¹H and ¹³C NMR and IR spectroscopy, and also by single crystal X-ray diffraction.Translated from Zhurnal Obshchei Khimii, Vol. 74, No. 11, 2004, pp. 1887–1893.Original Russian Text Copyright © 2004 by E. Klimova, Lorez, T. Klimova, Garcia, Hernandez, Ramirez.This revised version was published online in April 2005 with a corrected cover date.     

Source apportionment of groundwater pollutants in Apulian agricultural sites using multivariate statistical analyses: case study of Foggia province

Background

Ground waters are an important resource of water supply for human health and activities. Groundwater uses and applications are often related to its composition, which is increasingly influenced by human activities.In fact the water quality of groundwater is affected by many factors including precipitation, surface runoff, groundwater flow, and the characteristics of the catchment area. During the years 2004-2007 the Agricultural and Food Authority of Apulia Region has implemented the project “Expansion of regional agro-meteorological network” in order to assess, monitor and manage of regional groundwater quality. The total wells monitored during this activity amounted to 473, and the water samples analyzed were 1021. This resulted in a huge and complex data matrix comprised of a large number of physical-chemical parameters, which are often difficult to interpret and draw meaningful conclusions. The application of different multivariate statistical techniques such as Cluster Analysis (CA), Principal Component Analysis (PCA), Absolute Principal Component Scores (APCS) for interpretation of the complex databases offers a better understanding of water quality in the study region.

Results

Form results obtained by Principal Component and Cluster Analysis applied to data set of Foggia province it’s evident that some sampling sites investigated show dissimilarities, mostly due to the location of the site, the land use and management techniques and groundwater overuse. By APCS method it’s been possible to identify three pollutant sources: Agricultural pollution 1 due to fertilizer applications, Agricultural pollution 2 due to microelements for agriculture and groundwater overuse and a third source that can be identified as soil run off and rock tracer mining.

Conclusions

Multivariate statistical methods represent a valid tool to understand complex nature of groundwater quality issues, determine priorities in the use of ground waters as irrigation water and suggest interactions between land use and irrigation water quality.

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Characterization of immobilized nuclease P1

To improve the efficiency of the use of nuclease P1, enzyme immobilization technology was applied using nuclease P1. Characterization of immobilized nuclease P1 on different supports was studied. The results showed that the optimum pH and temperature of nuclease P1 immobilized via different supports were enhanced. The immobilized enzyme was obviously stable when stored for long periods and was reusable. The best results were obtained when nuclease P1 was immobilized on chitosan nanoparticles. The nanoparticles were applied to protect the activity of nuclease P1 and improved enzyme activity by 13.17% over that of free nuclease P1 at the same conditions. The Michaelis constant Km and V _max were determined for free and immobilized enzyme as well.     

Sensitive measurement of ultratrace phenols in natural water by purge-and-trap with in situ acetylation coupled with gas chromatography-mass spectrometry

A novel purge-and-trap method coupled with gas chromatography-mass spectrometry (GC-MS) is developed for the analysis of trace and ultratrace phenols based on their derivatization with acetic anhydride. Parameters affecting the extraction efficiency, such as purge temperature, concentration of sodium chloride, purge time, and volume of derivatization reagent, were investigated. The optimized conditions were addition of 150 μL acetic anhydride, purge time of 25 min at the purge temperature of 60 °C with 30% NaCl. The linear range was 0.2–100 μg L⁻¹ for phenols. The limits of detection (LODs) ranged from 0.08 to 0.15 μg L⁻¹ and the relative standard deviations (RSDs) for most of the phenols at the 10 μg L⁻¹level were below 10%. Natural water samples collected from a pool were successfully analyzed using the proposed method. The recovery of spiked water samples was 72.9–84.2%.