Dual Gradient Research Articles

Abstract B142: Dissociation of preclinical primary human cancer xenografts for cell surface transportome profiling.

Abstract Background: Tumor cell metabolism is of growing interest in both basic and clinical cancer research. A better understanding of underlying molecular mechanisms involving metabolite transport in normal and tumor cells should help drug discovery and development. Specific exofacial ligands to metabolite transporters derived from the receptor binding domain (RBD) of retrovirus envelope proteins were developed and used to quantify cell surface metabolite transporters. While cell surface labelling can be readily performed on cultured cell lines, analysis of single cells from solid tumors is more challenging. In this study, we developed a robust method for the disaggregation of tumor cells from human breast cancers grown as xenografts in mice. This procedure was then used to analyse the expression profiles of 4 cell membrane metabolite transporters involved in cell proliferation and tumorigenesis: Glut1, ASCT2, PiT1, and PiT2. Materials and methods: Eight primary human breast cancer xenografts were used for ex vivo experiments (Marangoni et al 2007). We developed an optimized disaggregation protocol to obtain maximum viable cell recovery from the xenografts. The protocol was validated for presence of CD44+ tumor cells and for cell viability using caspase 3 and DAPI exclusion and subsequently, applied to the xenografts for flow cytometry analyses, immunohistochemistry and ex vivo cell culture. Expression profiles of 4 metabolite transporters were assessed in 5 different human breast cancer xenografts. Results: The optimal dissociation protocol developed for these tumors combined mild non-enzymatic (non-enzymatic dissociation buffer) and enzymatic (collagenase III/DNase I) steps, followed by cell purification on a dual density Ficoll gradient. Less than 10% of resulting DAPI negative tumor cells were caspase 3 positive. Dissociated cells showed sustained viability in in vitro cultures for at least 12 days. The numbers of CD44+ cells determined by flow cytometry corresponded to those observed by IHC. The expression profiles of Glut1, ASCT2, PiT1, and PiT2 were distinct for each of the five human breast cancers, and metabolite transporter profiles were highly conserved for xenografts derived from the same tumor. Conclusions: Mouse xenograft implants of human breast cancer tumors were used to optimize and validate a dissociation method for the production of viable single cells. Cell suspensions were then assessed for cell surface metabolite transporters expression by flow cytometry. The expression patterns of four metabolite transporters, Glut1, ASCT2, PiT1, and PiT2 showed distinctive signature profiles for each group of xenografts, indicative of specific metabolic adaptations that can be tracked with our ligands for each tumor. Reference: 1. Marangoni E et al, CCR 2007;13:3989–3998. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference: Molecular Targets and Cancer Therapeutics; 2011 Nov 12-16; San Francisco, CA. Philadelphia (PA): AACR; Mol Cancer Ther 2011;10(11 Suppl):Abstract nr B142.

Value of hippocampal magnetic resonance spectroscopy in temporal lobe epilepsy disease

Objective To study the application value of hippocampus proton magnetic resonance spectroscopy (1H-MRS) in temporal lobe epilepsy. Methods With Philips Achieva 1.5T dual gradient MR scanner, 38 patients with temporal lobe epilepsy and 20 healthy volunteers were underwent brain MRI and hippocampal 1H-MRS examination, observation indicators included the concentration of NAA, Cr, Cho and the value of NAA/(Cho+Cr), NAA/Cr and NAA/Cho. The NAA/(Cho+Cr), NAA.Cr and NAA/Cho values in hippocampus of epilepsy side of epilepsy group, the normal side of epilepsy group, and the normal control group had a t test, analyzing the statistical difference between the two groups.Results The NAA/(Cho+Cr) values in hippocampal of the epilepsy side of epileptic patients, the normal side of epileptic patients and the control group were (0.42+0.06), (0.67+0.09), (0.72+0.07), respectively, there was significant difference between the epilepsy side and the normal side of epileptic patients, and between the epilepsy side of epileptic patients and the control group(P＜0.05), but no significant difference between the normal side of epileptic patients and the control group(P＞0.05). Using NAA/(Cho+Cr) to detect epilepsy, the sensitivity curve showed low→high→low changes, the highest sensitivity appeared in 0.60.Conclusion 1H-MRS not only has early diagnosis to hippocampal sclerosis, but also improves the accuracy of preoperative localization of epileptic focus. Key words: Hippocampal; Magnetic Resonance; Proton Magnetic Resonance Spectroscopy; Epilepsy

Experimental Evaluation of Separation Methods for a Riser Dilution Approach to Dual Density Drilling

A dual gradient, deepwater drilling system based on dilution of riser mud requires economically separating the riser mud into a low density dilution fluid and a higher density drilling fluid. This study investigated the practicality of accomplishing this separation using hydrocyclones and centrifuges and examined the possible benefits and efficiency of each. The separation experiments were conducted using a laboratory centrifuge and 2 in. hydrocyclones. The laboratory centrifuge was able to separate the riser mud into near ideal densities for dilution and drilling fluid. However, the dense slurry retained in the centrifuge had lower electrical stability than the feed stream. The hydrocyclones achieved much less contrast in density between the low and high density discharges, but their use consistently resulted in a beneficial increase in the stability of the mud emulsion in all of the flow streams and gave more desirable rheological properties. A qualitative comparison indicates that the hydrocyclone separation system may offer a feasible and desirable alternative to a centrifuge separation system.

Effectiveness of a three-dimensional dual gradient echo two-point Dixon technique for the characterization of adrenal lesions at 3 Tesla

To compare the sensitivity, specificity, and diagnostic accuracy of fat-only datasets reconstructed using a two-point Dixon technique, with corresponding opposed-phase (OP) and in-phase (IP) datasets for characterization of adrenal lesions at 3 Tesla (T). Fifty-nine patients (21 male, 38 female) with 66 adrenal lesions (49 adenomas, 17 nonadenomas) underwent 3D dual gradient-echo 3-T adrenal MR imaging with reconstruction of OP/IP and fat/water datasets. Sensitivity, specificity, and diagnostic accuracy were compared between OP/IP datasets, using the signal intensity index (SII), and fat/water datasets, using the fat fraction and fat ratio. Four radiologists qualitatively assessed OP/IP and fat-only datasets for reader confidence in lesion characterization and image quality. There were significant differences between adenomas and nonadenomas with regard to mean SII, fat fraction, and fat ratio (P < 0.001). There was no significant difference in mean diagnostic accuracy among different evaluation methods using OP/IP and fat/water datasets. Mean readers' scores for lesion characterization were significantly higher for adenomas than for nonadenomas using OP/IP and fat-only datasets. There was no significant difference between the two datasets regarding mean readers' scores for image quality. Fat-only images can readily differentiate adrenal adenomas from nonadenomas, with diagnostic accuracy comparable to OP/IP images.

Magnetic Resonance Imaging Estimation of Longitudinal Relaxation Rate Change (ΔR1) in Dual Gradient Echo Sequences Using an Adaptive Model.

Magnetic Resonance Imaging (MRI) estimation of contrast agent concentration in fast pulse sequences such as Dual Gradient Echo (DGE) imaging is challenging. An Adaptive Neural Network (ANN) was trained with a map of contrast agent concentration estimated by Look-Locker (LL) technique (modified version of inversion recovery imaging) as a gold standard. Using a set of features extracted from DGE MRI data, an ANN was trained to create a voxel based estimator of the time trace of CA concentration. The ANN was trained and tested with the DGE and LL information of six Fisher rats using a K-Fold Cross-Validation (KFCV) method with 60 folds and 10500 samples. The Area Under the Receiver Operator Characteristic Curve (AUROC) for 60 folds was used for training, testing and optimization of the ANN. After training and optimization, the optimal ANN (4:7:5:1) produced maps of CA concentration which were highly correlated (r =0.89, P < 0.0001) with the CA concentration estimated by the LL technique. The estimation made by the ANN had an excellent overall performance (AUROC = 0.870).

High Throughput Isolation and Glycosylation Analysis of IgG–Variability and Heritability of the IgG Glycome in Three Isolated Human Populations

All immunoglobulin G molecules carry N-glycans, which modulate their biological activity. Changes in N-glycosylation of IgG associate with various diseases and affect the activity of therapeutic antibodies and intravenous immunoglobulins. We have developed a novel 96-well protein G monolithic plate and used it to rapidly isolate IgG from plasma of 2298 individuals from three isolated human populations. N-glycans were released by PNGase F, labeled with 2-aminobenzamide and analyzed by hydrophilic interaction chromatography with fluorescence detection. The majority of the structural features of the IgG glycome were consistent with previous studies, but sialylation was somewhat higher than reported previously. Sialylation was particularly prominent in core fucosylated glycans containing two galactose residues and bisecting GlcNAc where median sialylation level was nearly 80%. Very high variability between individuals was observed, approximately three times higher than in the total plasma glycome. For example, neutral IgG glycans without core fucose varied between 1.3 and 19%, a difference that significantly affects the effector functions of natural antibodies, predisposing or protecting individuals from particular diseases. Heritability of IgG glycans was generally between 30 and 50%. The individual's age was associated with a significant decrease in galactose and increase of bisecting GlcNAc, whereas other functional elements of IgG glycosylation did not change much with age. Gender was not an important predictor for any IgG glycan. An important observation is that competition between glycosyltransferases, which occurs in vitro, did not appear to be relevant in vivo, indicating that the final glycan structures are not a simple result of competing enzymatic activities, but a carefully regulated outcome designed to meet the prevailing physiological needs.

Migration and maturation pattern of fetal enteric ganglia: A study of 16 cases

To study the migration and developmental pattern of ganglion cells in fetuses aged 9-21 weeks, and to document whether the migration was occurring circumferentially equally in the entire axis or if there were discrepancies in different portions at the same level. The hypothesis regarding the pathogenesis of Hirschsprung's disease mainly revolves around two schools. One is the single gradient migration of ganglia and the other is a dual gradient migration theory. Understanding the embryological development of enteric ganglia is necessary to study the pathogenesis of intestinal innervation disorders. We studied the development of intestinal ganglia in fetuses aged 9-21 weeks. Serial longitudinal sections from the colon were studied, the first one including the squamo-columnar junction, for the presence and the nature of ganglion cells with Hematoxylin and Eosin, and neurone-specific enolase immunostaining. Transverse sections from proximal gut were studied in a similar fashion. Thus, we evaluated the migration pattern as well as the nature of ganglia in the fetuses. We also measured the length of distal aganglionic segment in these growing fetuses. We noted that ganglion cells appear first in the myenteric plexus followed by deep and superficial submucous plexus. We also found evidences in favor of dual migration theory, and the distal aganglionic segment varies around the circumference of the rectal wall. We got evidences in support of a dual migration pattern of intestinal ganglion cells. The level of distal aganglionic segments when measured from squamo-columnar junction varied with the age of gestation and the length was incongruous. The description of distal aganglionic segment may help surgeons while taking biopsies or during operative procedures.

Feasibility of Texture Analysis for the Assessment of Biochemical Changes in Meniscal Tissue on T1 Maps Calculated From Delayed Gadolinium-Enhanced Magnetic Resonance Imaging of Cartilage Data

To (1) establish the feasibility of texture analysis for the in vivo assessment of biochemical changes in meniscal tissue on delayed gadolinium-enhanced magnetic resonance imaging of cartilage (dGEMRIC), and (2) compare textural with conventional T1 relaxation time measurements calculated from dGEMRIC data ("T1(Gd) relaxation times"). We enrolled 10 asymptomatic volunteers (7 men and 3 women; mean age, 27.2 +/- 4.5 years), without a history of meniscus damage, in our study. MRI of the right knee was performed at 3.0 T. An isotropic, 3-dimensional (3D), double-echo steady-state sequences was used for morphologic evaluation, and a dual flip angle 3D gradient echo sequence was used for T1(Gd) mapping. All MRI scans were performed 90 minutes after injection of 0.2 mmol/kg of Gd-diethylenetriamine pentaacetic acid (DTPA), and subsequently, during application of a compressive force (50% of the body weight) in the axial direction. Regions of interest, covering the central portions of the posterior horn of the medial meniscus, were defined on 3 adjacent sagittal sections. Based on the relaxation time maps, mean T1(Gd), as well as the T1(Gd) texture features derived from the co-occurrence matrix (COC: Angular Second Moment, Entropy, Inverse Difference Moment) and wavelet transform (WAV: WavEnLL, WavEnHL, WavEnHH, WavEnLH), were calculated. Paired t tests were used to assess differences between baseline and compression, and intraclass correlation coefficients (ICC) were calculated to establish the intrarater reliability of the measurements. Mean T1(Gd) (-67.3 ms, P = 0.011), Angular Second Moment (-0.0002, P = 0.009), Entropy (+0.033, P = 0.025), WavEnLL (+1011.16, P = 0.002), WavEnHL (+18.64, P = 0.012), and WavEnLH (+72.74, P = 0.035) differed significantly between baseline and compression. Intrarater reliability was substantial for mean T1(Gd) relaxation times (ICC = 0.99-1.0), and also for T1(Gd) co-occurrence matrix (ICC = 0.63-0.92) and WAV (ICC = 0.86-0.98) features. Texture features extracted from T1 maps calculated from dGEMRIC data are feasible for the in vivo assessment of biochemical changes in the menisci, such as might be induced by mechanical loading. Thus, T1(Gd) texture features complement conventional relaxation time measurements. Further studies are necessary to determine whether the mechanical compression, or a prolonged Gd-DTPA uptake, or both, are responsible for the observed decrease in mean T1(Gd) relaxation times in the menisci.

Stability of primal–dual gradient dynamics and applications to network optimization

This paper considers dynamic laws that seek a saddle point of a function of two vector variables, by moving each in the direction of the corresponding partial gradient. This method has old roots in the classical work of Arrow, Hurwicz and Uzawa on convex optimization, and has seen renewed interest with its recent application to resource allocation in communication networks. This paper brings other tools to bear on this problem, in particular Krasovskii’s method to find Lyapunov functions, and recently obtained extensions of the LaSalle invariance principle for hybrid systems. These methods are used to obtain stability proofs of these primal–dual laws in different scenarios, and applications to cross-layer network optimization are exhibited.

Online Dual Gradient Reversed-Phase/Porous Graphitized Carbon nanoHPLC for Proteomic Applications

The analysis of proteolytic peptide mixtures is among the dominant tasks within proteomic workflows. In order to limit undersampling effects during mass spectrometric detection, online-coupled liquid chromatography is the method of choice, with reversed-phase chromatography being the most important separation mode. Since hydrophilic compounds such as short peptides and some glycosylated species as well as oligosaccharides from glycoproteomic workflows are commonly not accessible by this analytical setup, we hereby present a dual gradient system combining reversed-phase and porous graphitic carbon retention modes within a single nanoHPLC setup. Samples in the low femtomole range are analyzed consecutively first by reversed phase, and nonretained molecules are directly separated by porous graphitic carbon. Both gradient elution systems allow for online coupled mass spectrometric detection and are demonstrated to enable analysis of protein, peptide, and oligosaccharide mixtures within the same setup. Thereby, the accessible range for proteomic and glycoproteomic applications may be extended far beyond the limits of conventional reversed-phase nanoHPLC setups.

Non-invasive assessment of hepatic steatosis: Prospective comparison of the accuracy of imaging examinations

Despite increasing use of various imaging examinations for non-invasive assessment of hepatic steatosis (HS), their relative accuracy is unknown. The objective of this study is to prospectively compare the accuracy of computed tomography (CT), dual gradient echo magnetic resonance imaging (DGE-MRI), proton magnetic resonance spectroscopy ((1)H-MRS), and ultrasonography (US) for the diagnosis and quantitative estimation of HS. A total of 161 consecutive potential living liver donors underwent US (performed by two independent radiologists, US1 and US2), CT, DGE-MRI, (1)H-MRS, and liver biopsy on the same day. Using the histologic degree of HS as the reference standard, we compared the diagnostic performance of US1, US2, CT, DGE-MRI, and (1)H-MRS for diagnosing HS >or= 5% and HS >or= 30% and compared the accuracy of CT, DGE-MRI, and (1)H-MRS in the quantitative estimation of HS. DGE-MRI and (1)H-MRS significantly outperformed CT and US for the diagnosis of HS5%. DGE-MRI showed a tendency of higher accuracy than the other examinations for diagnosing HS >or= 30%. The cross-validated sensitivity and specificity of DGE-MRI at the optimal cut-off were 76.7% and 87.1%, respectively, for diagnosing HS >or= 5% and 90.9% and 94%, respectively, for diagnosing HS >or= 30%. The cross-validated Bland-Altman 95% limits of agreement between the estimated degree of HS on imaging examinations and the histologic degree of HS, were the narrowest with DGE-MRI, yielding -12.7% to 12.7%. Among CT, DGE-MRI, (1)H-MRS, and US, DGE-MRI is the most accurate method for the diagnosis and quantitative estimation of HS. Therefore, DGE-MRI may be the preferred imaging examination for the non-invasive assessment of HS.

Spatial Composition Grading of Quaternary ZnCdSSe Alloy Nanowires with Tunable Light Emission between 350 and 710 nm on a Single Substrate

We demonstrated a general methodology of growing spatially composition-controlled alloys by combining spatial source reagent gradient with a temperature gradient. Using this dual gradient method, we achieved for the first time a continuous spatial composition grading of single-crystal quaternary Zn(x)Cd(1-x)S(y)Se(1-y) alloy nanowires over the complete band gap range along the length of a substrate. The band gap grading spans between 3.55 eV (ZnS) and 1.75 eV (CdSe) on a single substrate, with the corresponding light emission over the entire visible spectrum. We also showed that the dual gradient method can be extended to achieve alloy composition control in two spatial dimensions. The unique material platform achieved will open a wide range of applications from color engineered display and lighting, full spectrum solar cells, multispectral detectors, or spectrometer on-a-chip to superbroadly tunable nanolasers. The growth methodology can be extended more generally to other alloy systems.

A superlinear space decomposition algorithm for constrained nonsmooth convex program

A class of constrained nonsmooth convex optimization problems, that is, piecewise C 2 convex objectives with smooth convex inequality constraints are transformed into unconstrained nonsmooth convex programs with the help of exact penalty function. The objective functions of these unconstrained programs are particular cases of functions with primal–dual gradient structure which has connection with V U space decomposition. Then a V U space decomposition method for solving this unconstrained program is presented. This method is proved to converge with local superlinear rate under certain assumptions. An illustrative example is given to show how this method works.

Rheostatic control of tryptic digestion in a microscale fluidic system

Integrated fluidic systems that unite bottom-up and top-down proteomic approaches have the potential to deliver complete protein characterization. To circumvent fraction collection, as is conducted in current blended approaches, a technique to regulate digestion efficiency in a flow-through system is required. The present study examined the concept of regulating tryptic digestion in an immobilized enzyme reactor (IMER), incorporating mixed solvent systems for digestion acceleration. Using ovalbumin, cytochrome c, and myoglobin as protein standards, we demonstrate that tryptic digestion can be efficiently regulated between complete digestion and no digestion extremes by oscillating between 45 and 0% acetonitrile in the fluid stream. Solvent composition was tuned using programmable solvent waveforms in a closed system consisting of the IMER, a sample delivery stream, a dual gradient pumping system and a mass spectrometer. Operation in this rheostatic digestion mode provides access to novel peptide mass maps (due to substrate unfolding hysteresis) as well as the intact protein, in a reproducible and stable fashion. Although cycle times were on the order of 90 s for testing purposes, we show that regulated digestion is sufficiently rapid to be limited by solvent switching efficiency and kinetics of substrate unfolding/folding. Thus, regulated digestion should be useful in blending bottom-up and top-down proteomics in a single closed fluidic system.

Interaction of Actin and the Chloroplast Protein Import Apparatus

Actin filaments are major components of the cytoskeleton and play numerous essential roles, including chloroplast positioning and plastid stromule movement, in plant cells. Actin is present in pea chloroplast envelope membrane preparations and is localized at the surface of the chloroplasts, as shown by agglutination of intact isolated chloroplasts by antibodies to actin. To identify chloroplast envelope proteins involved in actin binding, we have carried out actin co-immunoprecipitation and co-sedimentation experiments on detergent-solubilized pea chloroplast envelope membranes. Proteins co-immunoprecipitated with actin were identified by mass spectrometry and by Western blotting and included the Toc159, Toc75, Toc34, and Tic110 components of the TOC-TIC protein import apparatus. A direct interaction of actin with Escherichia coli-expressed Toc159, but not Toc33, was shown by co-sedimentation experiments, suggesting that Toc159 is the component of the TOC complex that interacts with actin on the cytosolic side of the outer envelope membrane. The physiological significance of this interaction is unknown, but it may play a role in the import of nuclear-encoded photosynthesis proteins.

Physiology and Morphology of Color-Opponent Ganglion Cells in a Retina Expressing a Dual Gradient of S and M Opsins

Most mammals are dichromats, having short-wavelength-sensitive (S) and middle-wavelength-sensitive (M) cones. Smaller terrestrial species commonly express a dual gradient in opsins, with M opsin concentrated superiorly and declining inferiorly, and vice-versa for S opsin. Some ganglion cells in these retinas combine S- and M-cone inputs antagonistically, but no direct evidence links this physiological opponency with morphology; nor is it known whether opponency varies with the opsin gradients. By recording from >3000 ganglion cells in guinea pig, we identified small numbers of color-opponent cells. Chromatic properties were characterized by responses to monochromatic spots and/or spots produced by mixtures of two primary lights. Superior retina contained cells with strong S+/M- and M+/S- opponency, whereas inferior retina contained cells with weak opponency. In superior retina, the opponent cells had well-balanced M and S weights, while in inferior retina the weights were unbalanced, with the M weights being much weaker. The M and S components of opponent cell receptive fields had approximately the same diameter. Opponent cells injected with Lucifer yellow restricted their dendrites to the ON stratum of the inner plexiform layer and provided sufficient membrane area (approximately 2.1 x 10(4) microm(2)) to collect approximately 3.9 x 10(3) bipolar synapses. Two bistratified cells studied were nonopponent. The apparent decline in S/M opponency from superior to inferior retina is consistent with the dual gradient and a model where photoreceptor signals in both superior and inferior retina are processed by the same postreceptoral circuitry.

Delayed gadolinium-enhanced MRI of cartilage (dGEMRIC) of cadaveric shoulders: Comparison of contrast dynamics in hyaline and fibrous cartilage after intraarticular gadolinium injection

Delayed gadolinium-enhanced magnetic resonance imaging of cartilage (dGEMRIC) is a novel method to investigate cartilaginous and fibrocartilaginous structures. To investigate the contrast dynamics in hyaline and fibrous cartilage of the glenohumeral joint after intraarticular injection of gadopentetate dimeglumine. Transverse T(1) maps were acquired on a 1.5T scanner before and after intraarticular injection of 2.0 mmol/l gadopentetate dimeglumine in five cadaveric shoulders using a dual flip angle three-dimensional gradient echo (3D-GRE) sequence. The acquisition time for the T(1) maps was 5 min 5 s for the whole shoulder. Measurements were repeated every 15 min over 2.5 hours. Regions of interest (ROIs) covering the glenoid cartilage and the labrum were drawn to assess the temporal evolution of the relaxation parameters. T(1) of unenhanced hyaline cartilage of the glenoid was 568+/-34 ms. T(1) of unenhanced fibrous cartilage of the labrum was 552+/-38 ms. Significant differences (P=0.002 and 0.03) in the relaxation parameters were already measurable after 15 min. After 2 to 2.5 hours, hyaline and fibrous cartilage still demonstrated decreasing relaxation parameters, with a larger range of the T(1)(Gd) values in fibrous cartilage. T(1) and triangle Delta R(1) values of hyaline and fibrous cartilage after 2.5 hours were 351+/-16 ms and 1.1+/-0.09 s(-1), and 332+/-31 ms and 1.2+/-0.1 s(-1), respectively. A significant decrease in T(1)(Gd) was found 15 min after intraarticular contrast injection. Contrast accumulation was faster in hyaline than in fibrous cartilage. After 2.5 hours, contrast accumulation showed a higher rate of decrease in hyaline cartilage, but neither hyaline nor fibrous cartilage had reached equilibrium.

Fundamental equation of the dual flow rate-solvent gradient elution in liquid chromatography

An alternative expression of the fundamental equation of multi-mode gradient elution involving simultaneous changes in mobile phase composition and flow rate is derived using simple kinetic arguments and graphic interpretation. The new expression consists of a system of two integral equations and provides an easy and direct way of predicting retention times under dual-mode gradient conditions.

Comparison of Dual and Thermal Gradient Recording Methods for Thermally Assisted Magnetic Recording

The comparison between the dual gradient recording method (DGRM) and the thermal gradient recording method (TGRM) for thermally assisted magnetic recording (TAR) were investigated using the Landau-Lifshitz-Gilbert equation, which adds a stochastic thermal field to an effective field. The magnetization patterns and read/write characteristics were discussed. It was clarified that the write performance of DGRM was superior to TGRM for achieving a high areal recording density.

Quantitative measurements of corticosteroids in ex vivo samples using on-line SPE-LC/MS/MS

Abnormal elevation of 11β-HSD1 activities in tissues, such as fat and brain, may contribute to the development of the abdominal obesity and Alzheimer disease, and the inhibition of 11β-HSD1 might be beneficial to the management of these diseases. To assess the effects of pharmacologic inhibitors of 11β-HSD1, we developed a fast LC/MS/MS method to quantify corticosteroids in minced tissue samples in the presence of 11β-HSD substrates. The novel on-line SPE-LC/MS/MS method was developed with dual binary gradient and a throughput of 4.5 min/sample. A total of six corticosteroids (cortisol, cortisone, corticosterone, dehydrocorticosterone, dexamethasone, and dehydrodexamethasone) were studied. The lower limit of quantitation from 0.40 to 11.4 fmol and 4.5 orders magnitude of dynamic range were obtained for these six compounds. Three novel enzymatic bi-products, all isomers of cortisol, were observed in the liver or fat samples. Two of them were identified by matching the HPLC retention times and MS/MS spectra with authentic compounds. The potential interferences of these isomers and their removal are discussed.