Ion Optical Optimization Method for an Ultrahigh Resolution Planar Multireflection Time-of-Flight Mass Analyzer Using the Hill Climbing Algorithm.

A novel ion optical optimization method for planar multireflection time-of-flight mass spectrometry (MR-TOFMS) is introduced in this paper. The multiparameters of the gridless mirror model, including geometric and voltage parameters, are automatically optimized using a self-made program created in SIMION 8.1. Combining with the hill climbing algorithm and parallel computing technique, this method substantially enhances optimization efficiency and accuracy. The fitting results demonstrated that the ion optical performance of the gridless mirror reached up to fourth-order isochronicity with respect to the energy spread and third-order isochronicity with respect to the spatial and angular spread. As a result, the gridless mirror model achieved an aberration limit resolution of 1.7 million under realistic ion beam conditions. Due to constraints of periodic lenses, the aberration limit resolution of the planar MR-TOFMS was optimized to 600k. These results indicate that the hill climbing algorithm is an effective method to search the optimal solutions in complex ion optical systems.

Online Detection of HCN in Humid Exhaled Air by Gas Flow-Assisted Negative Photoionization Mass Spectrometry.

Hydrogen cyanide (HCN) is a well-known toxic compound in many fields. The trace amount of endogenous HCN in human exhalation has been associated with the presence of Pseudomonas aeruginosa (PA) infection in cystic fibrosis (CF) patients. Online monitoring of HCN profile is promising to screen PA infection rapidly and accurately. In this study, a gas flow-assisted negative photoionization (NPI) mass spectrometry method was developed for monitoring the single-exhalation HCN profile. The sensitivity could be optimized by introducing helium to eliminate the humidity influence and reduce the low mass cutoff effect, with improvements of a factor 150 observed. By employing a purging gas procedure and minimizing the length of the sample line, the residual and response time were greatly reduced. The limit of detection (LOD) of 0.3 ppbv and time resolution of 0.5 s were achieved. HCN profiles of exhalations from different volunteers before or after gargling with water were detected to show the performance of the method. All profiles showed a sharp peak and a stable end-tidal plateau, representing the concentration of oral cavity and end-tidal gas, respectively. The HCN concentration based on the plateau of the profile showed better reproducibility and accuracy, which indicates this method has potential application in the detection of PA infection in CF patients.

Rapid Determination of Ethyl Carbamate in Chinese Liquor via a Direct Injection Mass Spectrometry with Time-Resolved Flash-Thermal-Vaporization and Acetone-Assisted High-Pressure Photoionization Strategy.

Ethyl carbamate (EC), a carcinogenic compound, is naturally produced in fermented foods and alcoholic beverages. Rapid and accurate measurement of EC is necessary and important for quality control and safety evaluation of Chinese liquor, a traditionally distilled spirit with the highest consumption in China, but it remains a great challenge. In this work, a direct injection mass spectrometry (DIMS) with time-resolved flash-thermal-vaporization (TRFTV) and acetone-assisted high-pressure photoionization (HPPI) strategy has been developed. EC was rapidly separated from the main matrix components, ethyl acetate (EA) and ethanol, by the TRFTV sampling strategy due to the retention time difference of these three compounds with large boiling point differences on the inner wall of a poly(tetrafluoroethylene) (PTFE) tube. Therefore, the matrix effect of EA and ethanol was effectively eliminated. The acetone-assisted HPPI source was developed for efficient ionization of EC through a photoionization-induced proton transfer reaction between EC molecules and protonated acetone ions. The accurate quantitative analysis of EC in liquor was achieved by introducing an internal standard method (ISM) using deuterated EC (d5-EC). As a result, the limit of detection (LOD) for EC was 8.88 µg/L with the analysis time of only 2 min, and the recoveries ranged from 92.3 to 113.1%. Finally, the prominent capability of the developed system was demonstrated by rapid determination of trace EC in Chinese liquors with different flavor types, exhibiting wide potential applications in online quality control and safety evaluation of not only Chinese liquors but also other liquor and alcoholic beverages.

Design method and construction of the Schwarzschild microscope with high numerical aperture for secondary ion mass spectrometry.

The Schwarzschild microscope is suitable for sample navigation in secondary ion mass spectrometry (SIMS) because of its advantages of a simple structure, large working distance, and good coordination with the ion extraction system. The high numerical aperture (NA) of the microscope significantly reduces diffraction effects, but the resulting high-order geometric aberrations seriously affect the imaging quality. In this paper, a novel design method of the Schwarzschild microscope with a high NA (0.47) was proposed. The aberration distributions and compensation methods were investigated through tolerance analysis. The results showed that the tilt and decenter tolerances become the dominant factors limiting the spatial resolution, which could only be improved by ensuring the alignment accuracy of mirrors. Finally, the spatial resolution of the microscope in the home-built SIMS reached 2.19 µm.

Influence of Pseudopotential Well Depth on the Performance of Hybrid Linear Ion Trap/Time-of-Flight Mass Spectrometry Driven by Square and Sinusoidal Waveform.

Quadrupole devices are widely used as ion analyzers and ion guides, which are usually driven by a sinusoidal waveform. Recently, these devices driven by a digital waveform have attracted much attention. Driven by different rf waveforms, the pseudopotential well depth of quadrupole devices is significantly different, which will affect their performance. However, there is a lack of comprehensive view on the performance differences of quadrupole devices or hybrid instrument driven by different rf waveforms. In this paper, the differences in mass range, resolution, sensitivity, and ion fragmentation of LIT/TOFMS driven by square and sinusoidal waveforms were investigated. Compared with the sinusoidal mode, the square mode had a wider mass range and better mass resolution. Toluene was used to study the difference in sensitivity and ion fragmentation between the two modes. The results showed that the sensitivity of the square mode was 1.7-2.5 times that of the sinusoidal mode at different ion densities. According to the sensitivity differences, it was inferred that the ion-trapping efficiency and ion capacity of LIT in the square mode were 2.0 times and 1.7 times those in sinusoidal mode. It was found that the square mode behaved "softer" and presented less fragmentation, and the proportion of fragment ions in the sinusoidal mode can reach 3.2 times that in the square mode at high ion density. Therefore, LIT/TOFMS driven by a square waveform shows greater potential in the application of the complex matrix system with a wide mass range.

A Strategy for Simultaneously Improving Resolution and Sensitivity of Hybrid Quadrupole Ion Trap/Time-of-Flight Mass Spectrometry Using Square Waveform Phase Modulation.

The hybrid quadrupole ion trap/time-of-flight mass spectrometry (QIT/TOFMS) device is popular because of its advantages of high sensitivity, high resolution, and MS/MS capability. However, the analytical performance of QIT/TOFMS is severely limited by the parameters of the ion trap, as QIT is typically used as a TOF pulser because the ion initial distributions of space, velocity, and angle change dynamically with the phase angle of rf voltage. In this work, a square waveform phase modulation strategy was proposed to eliminate the influence of the rf phase angle, and the dependence of QIT/TOFMS performance on the phase angle was studied. It was found that the mass resolution and signal intensity showed a "W" trend with the increase of the ion extraction phase angle from 0° to 360°, where the best resolution and sensitivity were obtained at 0°, 180°, and 360° while the worst resolution and sensitivity were obtained at 90° and 270°. Moreover, the optimum phase angle was independent of m/z. As a result, the mass resolution for m/z 106, 164, and 258 was improved by 162%, 160%, and 210% respectively, while the signal intensities for m/z 106, 164, and 258 were enhanced by 25 ± 1, 10 ± 1, and 21 ± 1 fold, respectively, and a limit of detection down to 0.015 ppbv for m/z 164 was obtained. The experiment results indicated that the square waveform phase modulation strategy could be used to simultaneously improve the resolution and sensitivity of QIT/TOFMS.

High Mass Resolution Multireflection Time-of-Flight Secondary Ion Mass Spectrometer.

Time-of-flight secondary ion mass spectrometry (TOF-SIMS) is popular because of its advantages of parallel m/z detection and less damage for unknown or rare samples compared to sector field instruments. However, the mass resolving power of conventional TOF-SIMS is limited by its relatively large energy spread and primary ion pulse width. In this work, a high mass resolution multireflection time-of-flight secondary ion mass spectrometer (MR-TOF-SIMS) was designed and constructed. Compared with conventional TOF-SIMS, the ion flight path of the MR-TOF-SIMS was extended from meters to subkilometers, and the mass resolving power reached to 87000 after an 80 cycles flight. A pair of symmetrically arranged ion orthogonal injection/ejection deflectors, which could eliminate the influence of fringing field and remove ions with a large energy spread, were proposed to further improve the mass resolving power in fewer flight cycles. A zircon standard sample sputtered by a 10 keV O2- beam was used to demonstrate the performance of the MR-TOF-SIMS instrument. As a result, the mass resolving power was up to 30000 only after 22 flight cycles. The 92Zr+ peak was significantly separated from the mass interference peaks of 91ZrH+, 90ZrH2+, 13CC6H7+, and C7H8+. The mass accuracies of Zr ions and their hydrides were better than 1.2 ppm. An ion transmission efficiency over 40% was achieved after 115 cycles.

Rapid and highly sensitive measurement of trimethylamine in seawater using dynamic purge-release and dopant-assisted atmospheric pressure photoionization mass spectrometry.

Trimethylamine (TMA) is ubiquitous in the marine systems and may affect atmospheric chemistry as a precursor and strong stabilizer of atmospheric secondary aerosol, influencing cloud formation. Rapid and accurate measurement of the concentration of TMA in seawater is challenging due to their polarity, aqueous solubility, volatility and existence at low concentrations in marine environments. In this study, a dopant-assisted atmospheric pressure photoionization time-of-flight mass spectrometry (DA-APPI-TOFMS) coupled with a dynamic purge-release method was developed for rapid and sensitive analysis of TMA in seawater. A novel three-zones ionization source has been developed for improving the ionization efficiency of analyte molecules and minimizing the influence of high-humidity of the sample gas, which allowed direct analysis of high-humidity (RH> 90%) gas samples from microbubble purging process by the mass spectrometer. At atmospheric pressure, the three-zones ionization source allows the use of high-speed purge gas to quickly purge all organic amines dissolved in the water into the gas phase, ensuring quantitative accuracy. The limit of quantification (LOQ) for TMA down to 0.1 µg L-1 was obtained in less than 2 min by consuming only 2 mL seawater sample. This method was applied for the determination of the concentrations of TMA in the coastal seawater to validate its practicability and reliability for analysis of trace amines in marine environments.

Ionogels of pseudogemini supra-amphiphiles in ethylammonium nitrate: Structures and properties.

Thermoreversible ionogels formed by pseudogemini surfactants were prepared in protic ionic liquid, ethylammonium nitrate (EAN). Gemini-type supra-amphiphiles were formed by single-chain surfactants and bola-type molecules in a 2:1M ratio. The structures of aggregates including polymorphous lamellar structures and fibrous networks constituted by multilayer lamellae were determined by optical microscopy observations, transmission electron microscopy (TEM) observations, small-angle X-ray scattering (SAXS) and X-ray diffraction (XRD) measurements. The mechanism of molecular arrangement in aggregates was proposed. Transformation temperatures of samples which are closely related to the stability of well-ordered molecules in aggregates, as well as the rheological properties of ionogels were investigated systematically. The work affords a new way to construct ionogel by using the supramolecular self-assembly of pseudogemini-type molecules, and brings new ideas for the future construction of ionogels.

Metallosurfactant Ionogels in Imidazolium and Protic Ionic Liquids as Precursors To Synthesize Nanoceria as Catalase Mimetics for the Catalytic Decomposition of H2 O2.

The gelation behavior of cationic surfactants with different counterions, Br- , [FeCl3 Br]- , and [CeCl3 Br]- , in imidazolium ionic liquids (ILs) and protic ethylammonium nitrate was investigated. Small-angle X-ray scattering measurements and freeze-fracture transmission electron microscopy observations revealed the lamellar phases of metallosurfactant ionogels. The characteristics of imidazolium ILs, including the size and type, have effects on metallosurfactant ionogel properties, such as transformation temperatures, interlayer spacing, and mechanical strength. Cubic fluorite structured cerium oxide nanoparticles (CeO2 NPs) were produced by using metallosurfactant ionogels as precursors. Cubic fluorite CeO2 exhibited good catalase mimetic activity toward H2 O2 to generate O2 , providing more multiple mimetic enzyme activities of CeO2 NPs for H2 O2 .

Ionogels of a Sugar Surfactant in Ionic Liquids.

Green and environmentally friendly ionogels formed by a sugar surfactant were prepared in two kinds of imidazolium-based ionic liquids. The phase transition from ribbon structures to lamellar structures induced by temperature and the transition mechanism were investigated in detail by means of freeze-fracture TEM and field-emission SEM observations, as well as small-angle X-ray scattering measurements. The rheological properties and tribological properties of two kinds of ionogels were systematically investigated. The difference in the lubricating properties and antiwear capability can be explained well by the mechanical and viscoelastic properties, as well as the different microstructures of samples destroyed by shear forces. This work provides a better understanding of the relationship between the structures, rheological properties, and tribological properties of ionogels.

IkappaB kinase alpha kinase activity is required for self-renewal of ErbB2/Her2-transformed mammary tumor-initiating cells.

NF-kappaB is constitutively active in many solid tumors, including breast cancer. However, the role of NF-kappaB in breast carcinogenesis is unknown. Ikkalpha(AA/AA) "knockin" mice in which activation of IkappaB kinase alpha (IKKalpha) is prevented by replacement of activation loop serines with alanines exhibit delayed mammary gland growth during pregnancy, because IKKalpha activity is required for cyclin D1 induction and proliferation of lobuloalveolar epithelial cells. Given the role of cyclin D1 in breast and mammary cancer, we examined involvement of IKKalpha in mammary carcinogenesis induced by oncogenes or a chemical carcinogen, 7,12-dimethylbenz[a]anthracene (DMBA). The Ikkalpha(AA) mutation retarded tumor development in response to either 7,12-dimethylbenzaanthracene or the MMTV-c-neu (ErbB2/Her2) transgene but had no effect on MMTV-v-Ha-ras-induced cancer, although both oncogenes rely on cyclin D1. Strikingly, primary Ikkalpha(AA/AA)/MMTV-c-neu carcinoma cells exhibited diminished self-renewal capacity, resulting in the inability to establish secondary tumors. Ikkalpha(AA/AA)/MMTV-c-neu carcinoma cells underwent premature senescence when cultured under conditions used for propagation of mammary gland stem cells. Thus, IKKalpha is not only a regulator of mammary epithelial proliferation, but is also an important contributor to ErbB2-induced oncogenesis, providing signals that maintain mammary tumor-initiating cells. IKKalpha may represent a novel and specific target for treatment of ErbB2-positive breast cancer.

I{kappa}B kinase (IKK){beta}, but not IKK{alpha}, is a critical mediator of osteoclast survival and is required for inflammation-induced bone loss.

Transcription factor, nuclear factor kappaB (NF-kappaB), is required for osteoclast formation in vivo and mice lacking both of the NF-kappaB p50 and p52 proteins are osteopetrotic. Here we address the relative roles of the two catalytic subunits of the IkappaB kinase (IKK) complex that mediate NF-kappaB activation, IKKalpha and IKKbeta, in osteoclast formation and inflammation-induced bone loss. Our findings point out the importance of the IKKbeta subunit as a transducer of signals from receptor activator of NF-kappaB (RANK) to NF-kappaB. Although IKKalpha is required for RANK ligand-induced osteoclast formation in vitro, it is not needed in vivo. However, IKKbeta is required for osteoclastogenesis in vitro and in vivo. IKKbeta also protects osteoclasts and their progenitors from tumor necrosis factor alpha-induced apoptosis, and its loss in hematopoietic cells prevents inflammation-induced bone loss.

Activation of IKKalpha target genes depends on recognition of specific kappaB binding sites by RelB:p52 dimers.

IkappaB Kinase (IKK)alpha is required for activation of an alternative NF-kappaB signaling pathway based on processing of the NF-kappaB2/p100 precursor protein, which associates with RelB in the cytoplasm. This pathway, which activates RelB:p52 dimers, is required for induction of several chemokine genes needed for organization of secondary lymphoid organs. We investigated the basis for the IKKalpha dependence of the induction of these genes in response to engagement of the lymphotoxin beta receptor (LTbetaR). Using chromatin immunoprecipitation, we found that the promoters of organogenic chemokine genes are recognized by RelB:p52 dimers and not by RelA:p50 dimers, the ubiquitous target for the classical NF-kappaB signaling pathway. We identified in the IKKalpha-dependent promoters a novel type of NF-kappaB-binding site that is preferentially recognized by RelB:p52 dimers. This site links induction of organogenic chemokines and other important regulatory molecules to activation of the alternative pathway.

A dual role for Ikk alpha in tooth development.

IKK alpha is a component of the I kappa B kinase (IKK) complex that plays a key role in the activation of NF-kappa B. In Ikk alpha mutant mice and mice expressing a transdominant negative mutant of I kappa B alpha (cI kappa B alpha Delta N), molars have abnormal cusps, indicating that Ikk alpha is involved in cusp formation through the NF-kappa B pathway. However, Ikk alpha mutant incisors also have an earlier phenotype where epithelium evaginates outward into the developing oral cavity rather than invaginating into the underlying mesenchyme. A similar evagination of epithelium was also observed in whisker development, suggesting that Ikk alpha contributes to the direction of epithelial growth during the early stages of development in many ectodermal appendages. Since cI kappa B alpha Delta N mice have normal incisor epithelial invagination, Ikk alpha's role appears to be NF-kappa B independent. Changes in Notch1, Notch2, Wnt7b, and Shh expression found in incisor epithelium of Ikk alpha mutants suggest that this NF-kappa B-independent function is mediated by Notch/Wnt/Shh signaling pathways.

NF-kappaB in mammary gland development and breast cancer.

Nuclear factor of kappaB (NF-kappaB) is a group of sequence-specific transcription factors that is best known as a key regulator of the inflammatory and innate immune responses. Recent studies of genetically engineered mice have clearly indicated that NF-kappaB is also required for proper organogenesis of several epithelial tissues, including the mammary gland. Mice have shown severe lactation deficiency when NF-kappaB activation is specifically blocked in the mammary gland. In addition, there are strong suggestions that NF-kappaB may play an important role in the etiology of breast cancer. Elevated NF-kappaB DNA-binding activity is detected in both mammary carcinoma cell lines and primary human breast cancer tissues.

Proteotyping of mammary tissue from transgenic and gene knockout mice with immunohistochemical markers: a tool to define developmental lesions.

Through the use of transgenic and gene knockout mice, several studies have identified specific genes required for the functional development of mammary epithelium. Although histological and milk protein gene analyses can provide useful information regarding functional differentiation, they are limited in their ability to precisely define the molecular lesions. For example, mice that carry a mutation in one of the subunits of the IkappaB kinase, IKKalpha, cannot lactate despite the presence of histologically normal alveolar compartment and the expression of milk protein genes. To further define and understand such lesions on a molecular level, we sought evidence for proteins that are differentially expressed during mammary gland development with a view to generating a tissue proteotype. Using database screens and immunohistochemical analyses, we have identified three proteins that exhibit distinct profiles. Here, using mouse models as test biological systems, we demonstrate the development and application of mammary tissue proteotyping and its use in the elucidation of specific developmental lesions. We propose that the technique of proteotyping will have wide applications in the analyses of defects in other mouse models.

The lymphotoxin-beta receptor induces different patterns of gene expression via two NF-kappaB pathways.

The lymphotoxin-beta receptor (LTbetaR) plays critical roles in inflammation and lymphoid organogenesis through activation of NF-kappaB. In addition to activation of the classical NF-kappaB, ligation of this receptor induces the processing of the cytosolic NF-kappaB2/p100 precursor to yield the mature p52 subunit, followed by translocation of p52 to the nucleus. This activation of NF-kappaB2 requires NIK and IKKalpha, while NEMO/IKKgamma is dispensable for p100 processing. IKKbeta-dependent activation of canonical NF-kappaB is required for the expression but not processing of p100 and for the expression of proinflammatory molecules including VCAM-1, MIP-1beta, and MIP-2 in response to LTbetaR ligation. In contrast, IKKalpha controls the induction by LTbetaR ligation of chemokines and cytokines involved in lymphoid organogenesis, including SLC, BLC, ELC, SDF1, and BAFF.

NF-kappaB in cancer: from innocent bystander to major culprit.

Nuclear factor of kappaB (NF-kappaB) is a sequence-specific transcription factor that is known to be involved in the inflammatory and innate immune responses. Although the importance of NF-KB in immunity is undisputed, recent evidence indicates that NF-kappaB and the signalling pathways that are involved in its activation are also important for tumour development. NF-kappaB should therefore receive as much attention from cancer researchers as it has already from immunologists.