Investigation of Phospholipid Differences in Valproic Acid-Induced Autistic Mouse Model Brain Using Mass Spectrometry Imaging.

Autism is a neurodevelopmental disorder for which the cause and treatment have yet not been determined. The polyunsaturated fatty acid (PUFA) levels change rapidly in the blood or cerebrospinal fluid of autistic children and PUFAs are closely related to autism spectrum disorder (ASD). This finding suggests that changes in lipid metabolism are associated with ASD and result in an altered distribution of phospholipids in cell membranes. To further understand ASD, it is necessary to analyze phospholipids in organs consisting of nerve cells, such as the brain. In this study, we investigated the phospholipid distribution in the brain tissue of valproic acid-induced autistic mice using matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI-MSI). Phospholipids including phosphatidylcholine, phosphatidylethanolamine, and phosphatidylserine were identified in each brain region and exhibited differences between the ASD and control groups. These phospholipids contain docosahexaenoic acid and arachidonic acid, which are important PUFAs for cell signaling and brain growth. We expect that the differences in phospholipids identified in the brain tissue of the ASD model with MALDI-MSI, in conjunction with conventional biological fluid analysis, will help to better understand changes in lipid metabolism in ASD.

Triboresistive Touch Sensing: Grid-Free Touch-Point Recognition Based on Monolayered Ionic Power Generators.

Recent growing pursuit of skin-mountable devices has been impeded by the complicated structures of most sensing systems, containing electrode grids, stacked multilayers, and even external power sources. Here, a type of touch sensing, termed "triboresistive touch sensing", is introduced for gridless touch recognition based on monolayered ionic power generators. A homogeneous monolayer, i.e., ionic poly(dimethylsiloxane) (PDMS), generates electricity based on the electric field generated by touch. Voltages generated at each corner of the ionic PDMS rely on resistance between touch points and each corner, ensuring recognition of the touch positions without the need for electrode grid layers and external power sources. With notable advantages of high transparency (96.5%), stretchability (539.1%), and resilience (99.0%) of the ionic PDMS, epidermal triboresistive sensing is demonstrated to express touch position and readily play a musical instrument. A gridless system of triboresistive sensing allows rearrangement of the touch sections according to a given situation without any physical modification, and thus easily completes consecutive missions of controlling position, orientation, and gripping functions of a robot.

Matrix-Assisted Laser Desorption/Ionization Mass Spectrometry Imaging of Phospholipid Changes in a Drosophila Model of Early Amyotrophic Lateral Sclerosis.

Amyotrophic lateral sclerosis (ALS) is a degenerative disease caused by motor neuron damage in the central nervous system, and it is difficult to diagnose early. Drosophila melanogaster is widely used to investigate disease mechanisms and discover biomarkers because it is easy to induce disease in Drosophila through genetic engineering. We performed matrix-assisted laser desorption/ionization-mass spectrometry imaging (MALDI-MSI) to investigate changes in phospholipid distribution in the brain tissue of an ALS-induced Drosophila model. Fly brain tissues of several hundred micrometers or less were sampled using a fly collar to obtain reproducible tissue sections of similar sizes. MSI of brain tissues of Drosophila cultured for 1 or 10 days showed that the distribution of phospholipids, including phosphatidylcholine (PC), phosphatidylethanolamine (PE), phosphatidic acid (PA), phosphatidylserine (PS), and phosphatidylinositol (PI), was significantly different between the control group and the ALS group. In addition, the lipid profile according to phospholipids differed as the culture time increased from 1 to 10 days. These results suggest that disease indicators based on lipid metabolites can be discovered by performing MALDI-MSI on very small brain tissue samples from the Drosophila disease model to ultimately assess the phospholipid changes that occur in early-stage ALS.

Fabrication of inverse opal photonic gel sensors on flexible substrates by transfer process.

We demonstrate a platform technology for transferring opal films and photonic gel films to flexible substrates. The conventional fabrication procedure for inverse opal photonic gel (IOPG) sensors comprises three major steps: 1) the self-assembly of polystyrene µ-spheres to an opal template film within a channel between the top and bottom substrates, 2) infiltration and photo-polymerisation of the monomer mixture, and 3) etching of the opal template. Owing to the low processing yield of the first step, it is difficult to fabricate multiple sensor arrays on a single substrate. In this study, an opal film is formed between two substrates with different surface polarities, and the film is separated by disassembling the two substrates. The opal film on a medium polar substrate is covered using a flexible polyethylene terephthalate (PET) film, and opal-templated photo-polymerisation is performed. Finally, the photonic gel with the opal template is transferred to the PET film, and the opal template is etched out. Using the platform technique, the fabrications of pH-responsive IOPG and temperature-responsive IOPG sensors on PET films are respectively demonstrated. In addition, the IOPG containing the copolymer of acrylamide and 3-acrylamidophenylboronic acid was found to be responsive to glucose at physiological pH. All three sensors were fabricated using the same transfer method, differing only in the composition of monomer mixtures, and they all showed excellent sensitivity and repeatability on PET substrates. Due to the advantageous feature of the transfer method, dual sensors of pH-responsive IOPG and temperature-responsive IOPG were sequentially fabricated on a single PET film.

Ar-gas cluster ion beam in ToF-SIMS for peptide and protein analysis.

Since Ar-gas cluster ion beams (Ar-GCIBs) have been introduced into time-of-flight secondary ion mass spectrometry (ToF-SIMS), there have been various attempts to analyze organic materials and biomolecules that require low-damage analysis and high sensitivity, because Ar-GCIBs allow soft ionization of large molecules such as peptides and proteins due to the low energy per atom. Here, the authors adopted the Ar-GCIB as a primary beam to detect proteins including human insulin, ubiquitin, and cytochrome C (molecular weights are 5808, 8564, and 12 327 Da, respectively). They have confirmed that the detection of the intact proteins was possible when the Ar-GCIB was used as a primary ion beam. In addition, they successfully identified each protein by analyzing the trypsin-digested peptides in myoglobin, cytochrome C, and bovine serum albumin. They also attempted on-surface enzymatic digestion to identify proteins on the surface of the Si wafer and obtained results identical to those of in-solution digestion. It is expected that the authors' on-surface digestion method can enable the application of ToF-SIMS for the analysis of proteins present in biological tissues.

MALDI-TOF Mass Spectrometry Based on Parylene-Matrix Chip for the Analysis of Lysophosphatidylcholine in Sepsis Patient Sera.

In this work, medical diagnosis of sepsis was conducted via quantitative analysis of lysophosphatidylcholine 16:0 (LPC 16:0) by using matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF) mass spectrometry based on a parylene-matrix chip. In the first step, specific mass peaks for the diagnosis of sepsis were searched by comparing MALDI-TOF mass spectra of sepsis patient sera with healthy controls and pneumonia patient sera. Two mass peaks at m/z = 496.3 and 518.3 were chosen as those that are specifically different for sepsis sera to compare with healthy controls and pneumonia patient sera. These mass peaks were identified to be protonated and sodium adducts of LPC 16:0 by using tandem mass spectra (MS2 and MS3) of purely synthesized LPC 16:0 and extracted LPC 16:0 from a healthy control and a sepsis patient. In the next step, a standard curve for LPC 16:0 for the quantitative analysis of LPC 16:0 with MALDI-TOF MS based on the parylene-matrix chip was prepared, and the statistical correlation to the LC-MS analysis results was demonstrated by using the Bland-Altman test and Passing-Bablok regression. Finally, MALDI-TOF MS based on the parylene-matrix chip was used for the quantification of LPC 16:0 with sera from patients with severe sepsis and septic shock (n = 143), pneumonia patients (n = 12), and healthy sera (n = 31). The sensitivity and the selectivity of medical diagnosis of sepsis was estimated to be 97.9% and 95.5% by using MALDI-TOF MS based on the parylene-matrix chip, respectively.

Biological tissue sample preparation for time-of-flight secondary ion mass spectrometry (ToF-SIMS) imaging.

Time-of-flight secondary ion mass spectrometry (ToF-SIMS) imaging is an analytical technique rapidly expanding in use in biological studies. This technique is based on high spatial resolution (50-100 nm), high surface sensitivity (1-2 nm top-layer), and statistical analytic power. In mass spectrometry imaging (MSI), sample preparation is a crucial step to maintaining the natural state of the biomolecules and providing accurate spatial information. However, a number of problems associated with temperature changes in tissue samples such as loss of original distribution due to undesired molecular migration during the sample preparation or reduced ionization efficiency make it difficult to accurately perform MSI. Although frozen hydrate analysis is the ideal sample preparation method to eliminate the effects of temperature, this approach is hindered by mechanical limitations. Alternatively, an adhesive-tape-supported mounting and freeze-drying preparation has been proposed. This paper provides a concise review of the sample preparation procedures, a review of current issues, and proposes efficacious solutions for ToF-SIMS imaging in biological research.

Study of Photocatalytic Nano-Particle Effects on the Low Temperature Plasma Ionization Mass Spectrometry.

Although the low temperature plasma mass spectrometry (LTP-MS) is widely used as an analysis tool for many biochemical samples, its application window is somehow limited to the analytes of low molecular mass and high volatility. For this reason, there have been attempts to enhance the ionization/desorption efficiencies with extra heating, for instance. In this study, another enhancement method was suggested using the photocatalytic nano-particles (NPs). In order to assess the NP effects on the LTP-MS, two fatty acid ethyl ester samples of ethyl myristate and ethyl palmitate were used, and the NP of titanium dioxide (TiO2) was mainly employed. The results showed that the signal intensities of the LTP-MS were largely increased with the TiO2 addition for both samples. In addition, the cholesterol sample was analyzed using the TiO2 assisted LTP-MS, also resulting in the enhancement of the signal intensity. The overall results inferred that the photocatalytic NP confirmed its role as an effective assist tool for the LTP-MS, especially suitable because of the facile method and the heat-free nature. Graphical Abstract ᅟ.

Molecular depth profiling on rat brain tissue sections prepared using different sampling methods.

Brain imaging using time-of-flight secondary ion mass spectrometry (ToF-SIMS) has been reported to produce the distorted biomolecular distributions due to the cholesterol-induced matrix effect when cholesterol migrates to the surface, particularly in white matter, which contains a high level of cholesterol. Frozen-hydrated analysis has been used to inhibit the movement of cholesterol in the brain. In this paper, the authors propose new sample preparation and drying methods that can be used to obtain accurate biomolecular images at room temperature, instead of frozen-hydrated analysis using liquid-nitrogen, which must be continuously supplied to maintain the sample at -160 °C during the experiment. The rat brain prepared by the tape-supporting method on a precooled (-20 °C) stainless steel plate was freeze-dried in a load-lock chamber of ToF-SIMS for about an hour and moved directly to the main chamber. Using this preparation method, the authors found that cholesterol did not migrate to the surface in the corpus callosum (white matter) of the rat brain and sulfatide-related signals obtained from the cerebellum were not reduced in white matter. Our tape-supporting and freeze-drying sampling method for brain tissues could be a useful tool to study important metabolites of neurodegenerative diseases.

Sample Preparation of Corn Seed Tissue to Prevent Analyte Relocations for Mass Spectrometry Imaging.

Corn seed tissue sections were prepared by the tape support method using an adhesive tape, and mass spectrometry imaging (MSI) was performed. The effect of heat generated during sample preparation was investigated by time-of-flight secondary mass spectrometry (TOF-SIMS) imaging of corn seed tissue prepared by the tape support and the thaw-mounted methods. Unlike thaw-mounted sample preparation, the tape support method does not cause imaging distortion because of the absence of heat, which can cause migration of the analytes on the sample. By applying the tape-support method, the corn seed tissue was prepared without structural damage and MSI with accurate spatial information of analytes was successfully performed. Graphical Abstract ᅟ.

Bacterial analysis by laser desorption ionization mass spectrometry on amorphous silicon.

Lipid profiling in nine bacterial species has been accomplished by laser desorption ionization mass spectrometry (LDI-MS) using amorphous silicon (a-Si) thin film with 100 nm thickness. Lipid ions could be generated by LDI on a-Si regardless of ion acquisition modes because of a thermal property of a-Si to govern laser-induced surface heating. In a comparative study of lipid profiling in Bacillus lichemiformis by LDI-MS and matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS), LDI-MS on a-Si shows a higher efficiency in lipid and lipopeptide detection than MALDI-MS. A total of 53 peaks of lipid ions generated by LDI on a-Si in both acquisition modes for m/z 400-1200 was 1.6 times more than that detected by MALDI-MS using three organic matrices-2,5-dihydroxybenzoic acid, 1,5-diaminonaphthalene, and 2,4,6-trihydroxyacetophenone monohydrate. Also, the authors demonstrate by mass spectrometry imaging (MSI) that LDI-MS provides high detection coverage through whole sample area. MSI results show the detection yield in LDI on a-Si is 94.8% calculated by counting the number of points detected in the analyte ion signal in a whole spot. It means that reproducible detection of lipid ions by LDI-MS is possible even if laser is randomly irradiated at any position within the bacterial sample area applied on a-Si. Lipid profiling by LDI-MS on a-Si was applied to bacterial differentiation of nine bacterial species conducted by performing principal component analysis. Nine bacterial species are successfully distinguishable from each other by LDI-MS lipid profiling.

Effects of isometric hip movements on electromyographic activities of the trunk muscles during plank exercises.

[Purpose] The purpose of this study was to investigate the effect of isometric hip adduction and abduction on trunk muscle activity during plank exercises. [Subjects and Methods] Nineteen healthy male subjects were recruited for this study. All subjects performed the traditional plank exercise (TP), plank exercise with isometric hip adduction (PHAD), and plank exercise with isometric hip abduction (PHAB) by using an elastic band. Electromyographic (EMG) activities of the internal oblique (IO) and external oblique (EO) were measured during the 3 plank exercises by using an Electromyography system. [Results] Internal oblique and external oblique muscle activities were significantly greater during plank exercise with isometric hip adduction and plank exercise with isometric hip abduction than during traditional plank exercise. Internal oblique and external oblique muscle activities did not differ between the plank exercise with isometric hip adduction and plank exercise with isometric hip abduction conditions. [Conclusion] These findings demonstrate that loaded isometric hip movements may be a useful strategy to increase trunk muscle activity during plank exercises.

Improved mass resolution and mass accuracy in TOF-SIMS spectra and images using argon gas cluster ion beams.

The popularity of argon gas cluster ion beams (Ar-GCIB) as primary ion beams in time-of-flight secondary ion mass spectrometry (TOF-SIMS) has increased because the molecular ions of large organic- and biomolecules can be detected with less damage to the sample surfaces. However, Ar-GCIB is limited by poor mass resolution as well as poor mass accuracy. The inferior quality of the mass resolution in a TOF-SIMS spectrum obtained by using Ar-GCIB compared to the one obtained by a bismuth liquid metal cluster ion beam and others makes it difficult to identify unknown peaks because of the mass interference from the neighboring peaks. However, in this study, the authors demonstrate improved mass resolution in TOF-SIMS using Ar-GCIB through the delayed extraction of secondary ions, a method typically used in TOF mass spectrometry to increase mass resolution. As for poor mass accuracy, although mass calibration using internal peaks with low mass such as hydrogen and carbon is a common approach in TOF-SIMS, it is unsuited to the present study because of the disappearance of the low-mass peaks in the delayed extraction mode. To resolve this issue, external mass calibration, another regularly used method in TOF-MS, was adapted to enhance mass accuracy in the spectrum and image generated by TOF-SIMS using Ar-GCIB in the delayed extraction mode. By producing spectra analyses of a peptide mixture and bovine serum albumin protein digested with trypsin, along with image analyses of rat brain samples, the authors demonstrate for the first time the enhancement of mass resolution and mass accuracy for the purpose of analyzing large biomolecules in TOF-SIMS using Ar-GCIB through the use of delayed extraction and external mass calibration.

Formation of gas-phase peptide ions and their dissociation in MALDI: insights from kinetic and ion yield studies.

Insights on mechanisms for the generation of gas-phase peptide ions and their dissociation in matrix-assisted laser desorption ionization (MALDI) gained from the kinetic and ion yield studies are presented. Even though the time-resolved photodissociation technique was initially used to determine the dissociation kinetics of peptide ions and their effective temperature, it was replaced by a simpler method utilizing dissociation yields from in-source decay (ISD) and post-source decay (PSD). The ion yields for a matrix and a peptide were measured by repeatedly irradiating a region on a sample and collecting ion signals until the sample in the region was completely depleted. Matrix- and peptide-derived gas-phase cations were found to be generated by pre-formed ion emission or by ion-pair emission followed by anion loss, but not by laser-induced ionization. The total number of ions, that is, matrix plus peptide, was found to be equal to the number of ions emitted from a pure matrix. A matrix plume was found to cool as it expanded, from around 800-1,000 K to 400-500 K. Dissociation of peptide ions along b/y channels was found to occur statistically, that is, following RRKM behavior. Small critical energy (E0 = 0.6-0.7 eV) and highly negative critical entropy (ΔS(‡) = -30 to -25 eu) suggested that the transition structure was stabilized by multiple intramolecular interactions.

Ovarian teratoma mimicking metastasis on I-131 scan : a case report.

The whole body I-131 scan is routinely performed in the postoperative treatment of patients with well-differentiated thyroid cancer. Accurate interpretation of whole body I-131 scan after thyroidectomy is critical to appropriate management of patients with thyroid cancer, to prevent unnecessary surgical removal or exposure to radioiodine. Unfortunately, false-positive uptakes in several other organs and their associated disease processes have been reported. We report a case of false-positive iodine uptake in the pelvic region with incidentally diagnosed mature cystic teratoma.