Infrared spectroscopic evidence supporting heterogeneous site binding models for humic substances

Infrared spectroscopy was used to corroborate predictions made by newly developed heterogeneous site binding models for humic substances. Experimental conditions to acquire the spectra of soil humic substances (humic and fulvic acid and a polysaccharide fraction) in an aqueous state using horizontal attenuated total reflectance Fourier transform infrared spectroscopy (HATR-FTIR) were established. Elimination of the water spectrum from that of the sample was achieved by spectral subtraction of the water peak at 2020 cm(-1). A KSCN internal standard with an absorption band at 2067 cm(-1) was used to verify the efficacy of the subtraction procedure. Spectral artifacts produced by the water spectrum subtraction and from contaminants within the humic materials have been identified. Three fulvic and one humic acid solution were examined in solutions of varying pH. Results show that the observed proportion of ionized carboxylate in relation to pH is consistent with models that assume electrostatic effects and a continuous distribution of proton association constants (log KH). The spectroscopic data were in accordance with calculations made using the generic humic and fulvic acid NICA-Donnan model parameters.     

Quantification of particle number emission factors for motor vehicles from on-road measurements

The database on particle number emission factors has been very limited to date despite the increasing interest in the effects of human exposure to particles in the submicrometer range. There are also major questions on the comparability of emission factors derived through dynamometer versus on-road studies. Thus, the aims of this study were (1) to quantify vehicle number emission factors in the submicrometer (and also supermicrometer) range for stop-start and free-flowing traffic at about 100 km h(-1) driving conditions through extensive road measurements and (2) to compare the emission factors from the road measurements with those obtained previously from dynamometer studies conducted in Brisbane. For submicrometer particles the average emission factors for Tora Street were estimated at (1.89 +/- 3.40) x 10(13) particles km(-1) (mean +/- standard error; n = 386) for petrol and (7.17 +/- 2.80) x 10(14) particles km(-1) (diesel; n = 196) and for supermicrometer particles at 2.59 x 10(9) particles km(-1) and 1.53 x 10(12) particles km(-1), respectively. The average number emission factors for submicrometer particles estimated for Ipswich Road (stop-start traffic mode) were (2.18 +/- 0.57) x 10(13) particles km(-1) (petrol) and (2.04 +/- 0.24) x 10(14) particles km(-1) (diesel). One implication of the conclusion that emission factors of heavy duty diesel vehicles are over 1 order of magnitude higher than emission factors of petrol-fueled passenger cars is that future control and management strategies should in particular target heavy duty vehicles, as even a moderate decrease in emissions of these vehicles would have a significant impact on lowering atmospheric concentrations of particles. The finding that particle number emissions per vehicle-km are significantly larger for higher speed vehicle operation has an important implication on urban traffic planning and optimization of vehicle speed to lower their impact on airborne pollution. Additionally, statistical analysis showed that neither the measuring method (dynamometer or on-road), nor data origin (Brisbane or elsewhere in the world), is associated with a statistically significant difference between the average values of emission factors for diesel, petrol, and vehicle fleet mix. However, statistical analyses of the effect of fuel showed that the mean values of emission factors for petrol and diesel are different at a 5% significance level.     

Non-targeted analysis of tea by hydrophilic interaction liquid chromatography and high resolution mass spectrometry

Tea is the second most consumed beverage in the world and its consumption has been associated with numerous potential health benefits. Factors such as fermentation methods, geographical origin and season can affect the primary and secondary metabolite composition of tea. In this study, a hydrophilic interaction liquid chromatography (HILIC) method coupled to high resolution mass spectrometry in both positive and negative ionisation modes was developed and optimised. The method when combined with principal component analysis to analyse three different types of tea, successfully distinguished samples into different categories, and provided evidence of the metabolites which differed between them. The accurate mass and high resolution attributes of the mass spectrometric data were utilised and relative quantification data were extracted post-data acquisition on 18 amino acids, showing significant differences in amino acid concentrations between tea types and countries. This study highlights the potential of HILIC chromatography combined with non-targeted mass spectrometric methods to provide a comprehensive understanding of polar metabolites in plant extracts.     

Deoxynivalenol exposure assessment in a cohort of pregnant women from Bradford, UK

Deoxynivalenol (DON) is a ubiquitous contaminant of cereal crops in temperate regions of the world. It causes growth faltering and immune suppression in animals. Limited information is available on DON exposure in UK subpopulations. The objective of this study was to provide DON exposure assessment in a subset of pregnant women scheduled for an elective caesarean in a large multi-ethnic mother/infant birth cohort from Bradford, UK. Women aged 16-44 years (n = 85) provided a urine sample for DON analysis in the last trimester of pregnancy, and concurrently completed a food-frequency questionnaire (FFQ). The urinary DON biomarker was detected in all measured samples (geometric mean (GM)?= 10.3 ng DON mg(-1) creatinine, range = 0.5-116.7 ng mg(-1)). Levels were higher in women classified as South Asian in origin (GM: 15.2 ng mg(-1); 95% CI = 10.7-21.5 ng mg(-1)) compared with non-South Asians (GM = 8.6 ng mg(-1); 95% CI = 6.6-11.8 ng mg(-1)), p = 0.02). Estimated DON intake from FFQ data and typical levels of DON contamination of food suggest that this was mainly due to higher levels of exposure from bread, particularly daily intake of DON from chapattis in South Asians (estimated mean = 2.4 μg day(-1); 95% CI = 1.2, 3.7 μg day(-1)) compared with non-South Asians (estimated mean = 0.2 μg day(-1); 95% CI = 0-0.4 μg day(-1)), p < 0.001. This is the first biomarker demonstration of DON exposure in pregnant women, and several urinary DON levels were the highest ever recorded in any study. A larger survey within this birth cohort is warranted to investigate any potential risk to mothers and their babies, from DON exposure during pregnancy.     

Precise measurements of intrauterine vascular structures at hysteroscopy in menorrhagia and during Norplant use

Using currently available equipment for panoramic hysteroscopy, the size of images viewed cannot be accurately judged because of the magnifying and distorting effects of the objective lens. This study has demonstrated that magnification by the hysteroscope lens can alter the apparent size of images by up to 27%. An additional effect of lens distortion can alter the apparent size of images viewed by up to 28%, depending on the position of the image in the field of view. These effects are independent and may be additive. Thus, the apparent size of intrauterine structures at hysteroscopy may bear little resemblance to their actual size. Image-correction methods are described which reduce the effects of image magnification on the apparent size of an object viewed through the hysteroscope to 7%, and the effect of distortion to 3 %. This technique can greatly improve the accuracy of measurement at hysteroscopy, and has been utilized in this study for the precise measurement of superficial endometrial vascular diameter (mean+/-SEM) in 34 Norplant users (120+/-11.6 microm) and 20 women with spontaneous menorrhagia (74+/-7.2 microm). It has also confirmed the presence of scattered dilated vessels (up to 777 microm in diameter) on the endometrial surface in some Norplant users.     

Arousal of negative emotions and symptom-specific reactivity in chronic low back pain patients.

Anger may have greater effects on chronic pain severity than other negative emotions and may do so by increasing muscle tension near the site of injury (symptom-specific reactivity). For patients with chronic low back pain (CLBP), relevant muscles are lower paraspinals (LP). Ninety-four CLBP patients and 79 controls underwent anger and sadness recall interviews. EMG and cardiovascular activity were recorded. Patients exhibited greater LP tension increases during anger and slower recovery than controls. Only patients showed greater LP reactivity during anger than sadness. For both groups, trapezius reactivity during anger and sadness did not differ. LP reactivity to anger correlated with everyday pain severity for patients. Anger-induced symptom-specific LP reactivity may be linked to chronic pain aggravation among CLBP patients. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Mechanized silica nanoparticles: a new frontier in theranostic nanomedicine

Medicine can benefit significantly from advances in nanotechnology because nanoscale assemblies promise to improve on previously established therapeutic and diagnostic regimes. Over the past decade, the use of delivery platforms has attracted attention as researchers shift their focus toward new ways to deliver therapeutic and/or diagnostic agents and away from the development of new drug candidates. Metaphorically, the use of delivery platforms in medicine can be viewed as the "bow-and-arrow" approach, where the drugs are the arrows and the delivery vehicles are the bows. Even if one possesses the best arrows that money can buy, they will not be useful if one does not have the appropriate bow to deliver the arrows to their intended location. Currently, many strategies exist for the delivery of bioactive agents within living tissue. Polymers, dendrimers, micelles, vesicles, and nanoparticles have all been investigated for their use as possible delivery vehicles. With the growth of nanomedicine, one can envisage the possibility of fabricating a theranostic vector that could release powerful therapeutics and diagnostic markers simultaneously and selectively to diseased tissue. In our design of more robust theranostic delivery systems, we have focused our attention on using mesoporous silica nanoparticles (SNPs). The payload "cargo" molecules can be stored within this robust domain, which is stable to a wide range of chemical conditions. This stability allows SNPs to be functionalized with stimulus-responsive mechanically interlocked molecules (MIMs) in the shape of bistable rotaxanes and psuedorotaxanes to yield mechanized silica nanoparticles (MSNPs). In this Account, we chronicle the evolution of various MSNPs, which came about as a result of our decade-long collaboration, and discuss advances in the synthesis of novel hybrid SNPs and the various MIMs which have been attached to their surfaces. These MIMs can be designed in such a way that they either change shape or shed off some of their parts in response to a specific stimulus, such as changes in redox potential, alterations in pH, irradiation with light, or the application of an oscillating magnetic field, allowing a theranostic payload to be released from the nanopores to a precise location at the appropiate time. We have also shown that these integrated systems can operate not only within cells, but also in live animals in response to pre-existing biological triggers. Recognizing that the theranostics of the future could offer a fresh approach to the treatment of degenerative diseases including cancer, we aim to start moving out of the chemical domain and into the biological one. Some MSNPs are already being tested in biological systems.     

Production of active recombinant eIF5A: reconstitution in E.coli of eukaryotic hypusine modification of eIF5A by its coexpression with modifying enzymes

Eukaryotic translation initiation factor 5A (eIF5A) is the only cellular protein that contains the polyamine-modified lysine, hypusine [N(ε)-(4-amino-2-hydroxybutyl)lysine]. Hypusine occurs only in eukaryotes and certain archaea, but not in eubacteria. It is formed post-translationally by two consecutive enzymatic reactions catalyzed by deoxyhypusine synthase (DHS) and deoxyhypusine hydroxylase (DOHH). Hypusine modification is essential for the activity of eIF5A and for eukaryotic cell proliferation. eIF5A binds to the ribosome and stimulates translation in a hypusine-dependent manner, but its mode of action in translation is not well understood. Since quantities of highly pure hypusine-modified eIF5A is desired for structural studies as well as for determination of its binding sites on the ribosome, we have used a polycistronic vector, pST39, to express eIF5A alone, or to co-express human eIF5A-1 with DHS or with both DHS and DOHH in Escherichia coli cells, to engineer recombinant proteins, unmodified eIF5A, deoxyhypusine- or hypusine-modified eIF5A. We have accomplished production of three different forms of recombinant eIF5A in high quantity and purity. The recombinant hypusine-modified eIF5A was as active in methionyl-puromycin synthesis as the native, eIF5A (hypusine form) purified from mammalian tissue. The recombinant eIF5A proteins will be useful tools in future structure/function and the mechanism studies in translation.     

Phosphorylation Stoichiometries of Human Eukaryotic Initiation Factors

Eukaryotic translation initiation factors are the principal molecular effectors regulating the process converting nucleic acid to functional protein. Commonly referred to as eIFs (eukaryotic initiation factors), this suite of proteins is comprised of at least 25 individual subunits that function in a coordinated, regulated, manner during mRNA translation. Multiple facets of eIF regulation have yet to be elucidated; however, many of the necessary protein factors are phosphorylated. Herein, we have isolated, identified and quantified phosphosites from eIF2, eIF3, and eIF4G generated from log phase grown HeLa cell lysates. Our investigation is the first study to globally quantify eIF phosphosites and illustrates differences in abundance of phosphorylation between the residues of each factor. Thus, identification of those phosphosites that exhibit either high or low levels of phosphorylation under log phase growing conditions may aid researchers to concentrate their investigative efforts to specific phosphosites that potentially harbor important regulatory mechanisms germane to mRNA translation.