A new method for wide frequency range dynamic olfactory stimulation and characterization

Sensory receptors often receive strongly dynamic, or time varying, inputs in their natural environments. Characterizing their dynamic properties requires control and measurement of the stimulus over a frequency range that equals or exceeds the receptor response. Techniques for dynamic stimulation of olfactory receptors have lagged behind other major sensory modalities because of difficulties in controlling and measuring the concentration of odorants at the receptor. We present a new method for delivering olfactory stimulation that gives linear, low-noise, wide frequency range control of odorant concentration. A servo-controlled moving bead of silicone elastomer occludes the tip of a Pasteur pipette that releases odorant plus tracer gas into a flow tube. Tracer gas serves as a surrogate indicator of odorant concentration and is measured by a photoionization detector. The system has well-defined time-dependent behavior (frequency response and impulse response functions) and gives predictable control of odorant over a significant volume surrounding the animal. The frequency range of the system is about 0-100 Hz. System characterization was based on random (white noise) stimulation, which allows more rapid and accurate estimation of dynamic behavior than deterministic signals such as sinusoids or step functions. Frequency response functions of Drosophila electroantennograms stimulated by fruit odors were used to demonstrate a typical application of the system.     

尾叶桉异戊二烯排放量初步研究

应用光离子化气相色谱仪(GC-PID)测定尾叶桉3月份的异戊二烯排放量,同时采用Li-6400光合作用测定仪同步测定叶片净光合速率、蒸腾速率、气孔导度和胞间CO2浓度,及其相关环境因子,并对截枝后尾叶桉异戊二烯排放量的变化进行分析。结果表明,春季尾叶桉异戊二烯白天的排放量变化呈单峰型曲线,最大值出现在14:00左右,截枝后尾叶桉的异戊二烯排放量下降。     

Temporal integrity of an airborne odor stimulus is greatly affected by physical aspects of the odor delivery system

There is currently a debate about the role played by temporal patterns in neural activity in olfactory coding. An accurate analysis of this question, however, is only possible if the temporal properties of a stimulus itself are well defined. So far, no technique with sufficient temporal resolution has been available to accomplish this. Using a photoionization detector (PID), we show that the configuration of the odor delivery apparatus and the airflow settings greatly influence the integrity of a stimulus profile within an odor delivery apparatus. In a situation where pulsatile odor stimuli are applied to a stationary preparation, we tested the effect of 1) axial and off-center location within the airstream, 2) airflow of the odor delivery, 3) exit tube length, 4) exit tube diameter, 5) orientation of the odor delivery device in relation to the exhaust flow, and 6) exhaust tube air speed. This has important implications for the study of time in olfaction; significant planning must be incorporated into the design of the experiment to provide a well-defined odor delivery system.     

Determination of 17 illicit drugs in oral fluid using isotope dilution ultra-high performance liquid chromatography/tandem mass spectrometry with three atmospheric pressure ionizations

The collection of oral fluid for drug testing is easy and non-invasive. This study developed a drug testing method using ultra-high performance liquid chromatography/tandem mass spectrometry (UHPLC-MS/MS) in selected-reaction monitoring (SRM) mode. We tested the method on the analysis of four opiates and their metabolites, five amphetamines, flunitrazepam and its two metabolites, and cocaine and its four metabolites in oral fluid. 100-μL samples of oral fluid were diluted with twice the amount of water then spiked with isotope-labeled internal standards. After the samples had undergone high-speed centrifugation for 20 min, we analyzed the supernatant. The recovery of the sample preparation ranged from 81 to 108%. We compared the performance of electrospray ionization (ESI), atmospheric pressure chemical ionization (APCI) and atmospheric pressure photoionization (APPI). The ion suppression of most analytes on ESI (28-78%) was lower than that of APCI and APPI. A post-column flow split (5:1) did not reduce the matrix effect on ESI. Direct APPI performed better than dopant-assisted APPI using toluene. ESI, APCI and APPI limits of quantitation mostly ranged from 0.11 to 1.9 ng/mL, 0.02 to 2.2 ng/mL and 0.02 to 2.1 ng/mL, respectively, but were much higher on amphetamine and ecgonine methyl ester (about 2.7-4.7 ng/mL, 8.7-14 ng/mL, and 10-19 ng/mL, respectively). Most of the bias percentages (accuracy) and relative standard deviations (precision) on spiked samples were below 15%. This method greatly simplifies the process of sample preparation and shortens the chromatographic time to only 7.5 min per run and is able to detect analytes at sub-ppb levels.     

Shake-up and shake-off excitations with associated electron losses in X-ray studies of proteins

Photoionization of an atom by X-rays usually removes an inner shell electron from the atom, leaving behind a perturbed "hollow ion" whose relaxation may take different routes. In light elements, emission of an Auger electron is common. However, the energy and the total number of electrons released from the atom may be modulated by shake-up and shake-off effects. When the inner shell electron leaves, the outer shell electrons may find themselves in a state that is not an eigen-state of the atom in its surroundings. The resulting collective excitation is called shake-up. If this process also involves the release of low energy electrons from the outer shell, then the process is called shake-off. It is not clear how significant shake-up and shake-off contributions are to the overall ionization of biological materials like proteins. In particular, the interaction between the outgoing electron and the remaining system depends on the chemical environment of the atom, which can be studied by quantum chemical methods. Here we present calculations on model compounds to represent the most common chemical environments in proteins. The results show that the shake-up and shake-off processes affect approximately 20% of all emissions from nitrogen, 30% from carbon, 40% from oxygen, and 23% from sulfur. Triple and higher ionizations are rare for carbon, nitrogen, and oxygen, but are frequent for sulfur. The findings are relevant to the design of biological experiments at emerging X-ray free-electron lasers.     

Chemiluminescence from luminol solution after illumination of a 355 nm pulse laser

Chemiluminescence (CL) on the time scale of microseconds to milliseconds from luminol solution after illumination of a 355 nm pulse laser is reported. It was found that the CL is the emission from 3‐aminophthalate ion (AP*). In CL decay after the pulse laser illumination, a peak was observed from about 200 to 30 µs depending on the laser power and the luminol concentration. It seemed that there was a fast and slow decay process; their kinetics were greatly dependent on the laser power and the luminol concentration. Dissolved oxygen was involved in the CL and played the same role on the whole time scale of microseconds to milliseconds. Involvement of reactive oxygen species such as H₂O₂, ¹O₂, O₂^?? and OH in the CL was examined by adding their scavengers. Experimental results suggested that the possibility of involvement of H₂O₂ and ¹O₂ in the CL was low. The CL in time periods less than 50 µs might be related to ?OH. The ^?O₂^??‐induced CL increased with time after 50 µs and became dominant on the time scale of milliseconds. The CL was considered to be caused by both the photoionization and type I reaction mechanisms. Copyright © 2009 John Wiley & Sons, Ltd.     

Atmospheric pressure photoionization mass spectrometry of per-O-methylated oligosaccharides related to D-xylans

Three d-xylan type per-O-methylated trisaccharides with various types of linkages between the d-xylopyranose units were examined by atmospheric pressure photoionization (APPI) mass spectrometry in the positive ion mode. The most interesting feature of a thermospray mass spectrum using the APPI source with UV lamp switched off, is the exclusive production of [M+Na]⁺ adduct ions. [M+Na]⁺ cationized ions are the most abundant species in the case of APPI mass spectrometry. The second ionization process has no analogy in the case of substances studied using APPI to date. This aspect involves the addition of a water molecule to the molecular ion of a per-O-methylated saccharide, giving rise to [M+H₂O]⁺ adduct ions. The [M+H₂O]⁺ species are readily detected at m/z 544, and are clearly visible for all three isomers studied. The MS/MS spectrum of [M+Na]⁺ ions contains a base peak at m/z 375, produced by a Y-type cleavage of the trisaccharide, along with a hydrogen rearrangement on the terminal interglycosidically linkage glycosidic oxygen atom. The [M+H₂O]⁺ species fragment largely give rise to ions at m/z 175, 143 and, as a result, the m/z 111 ion is unique to nonreducing terminal units.     

Correlations between PID and FID Field Analytical Instruments in the Analysis of Soil Contaminated with Diesel Fuel

Research was conducted to determine if there is a correlation between the data gathered by field analytical instruments in analyzing soil contaminated with diesel fuel. One instrument was equipped with a flame ionization detector (FID) and the other a photoionization detector (PID). The results showed that the concentration readings of the PID and FID displayed a linear relationship for soil recently contaminated with diesel fuel. However, for soil containing weathered diesel fuel in the field, a logarithmic relationship between the PID and FID readings was displayed. It was also determined by laboratory experimentation that the PID and FID readings both exhibited log-linear decreases over time for uncovered diesel fuel-contaminated soil. It was concluded that the PID and FID can both individually be used to evaluate soil contaminated by diesel fuel and might be interchangeable depending on the needs of the researcher.     

Sensitive UHPLC-MS/MS quantification method for 4β- and 4α-hydroxycholesterol in plasma for accurate CYP3A phenotyping

4β-Hydroxycholesterol (4β-OHC) is formed by Cytochrome P450 (CYP)3A and has drawn attention as an endogenous phenotyping probe for CYP3A activity. However, 4β-OHC is also increased by cholesterol autooxidation occurring in vitro due to dysregulated storage and in vivo by oxidative stress or inflammation, independent of CYP3A activity. 4α-hydroxycholesterol (4α-OHC), a stereoisomer of 4β-OHC, is also formed via autooxidation of cholesterol, not by CYP3A, and thus may have clinical potential in reflecting the state of cholesterol autooxidation. In this study, we establish a sensitive method for simultaneous quantification of 4β-OHC and 4α-OHC in human plasma using ultra-high performance liquid chromatography coupled to tandem mass spectrometry. Plasma samples were prepared by saponification, two-step liquid-liquid extraction, and derivatization using picolinic acid. Intense [M+H]+ signals for 4β-OHC and 4α-OHC di-picolinyl esters were monitored using electrospray ionization. The assay fulfilled the requirements of the US Food and Drug Administration guidance for bioanalytical method validation, with a lower limit of quantification of 0.5 ng/ml for both 4β-OHC and 4α-OHC. Apparent recovery rates from human plasma ranged from 88.2% to 101.5% for 4β-OHC, and 91.8% to 114.9% for 4α-OHC. Additionally, matrix effects varied between 86.2% and 117.6% for 4β-OHC and between 89.5% and 116.9% for 4α-OHC. Plasma 4β-OHC and 4α-OHC concentrations in healthy volunteers, stage 3–5 chronic kidney disease (CKD) patients, and stage 5D CKD patients as measured by the validated assay were within the calibration ranges in all samples. We propose this novel quantification method may contribute to accurate evaluation of in vivo CYP3A activity.     

激光所致核酸损伤的分子机制

本文综述了近来有关紫外激光诱导核酸损伤(主要是碱基损伤、单链断裂和双链断裂)形成的化学过程的文献报导,主要是讨论了双光子激发核酸碱基,致使核酸产生单链断裂和双链断裂的光致电离过程,以及由此归纳出来的激光诱导核酸链断裂的化学模型