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Unique functional materials provide a platform as scaffolds for cell/tissue regeneration. Investigation of cell-materials' chemical and biological interactions will enable the application of more functional materials in the area of bioengineering, which provides a pathway to the novel treatment for patients who suffer from tissue/organ damage and face the limitation of donation sources. Many studies have been made into tissue/organ regeneration. Development of new substrate materials as platforms for cell/tissue regeneration is a key research area. Studies discussed in this paper focus on the investigation of novel ultrananocrystalline diamond (UNCD) films as substrate/scaffold materials for developmental biology. Specially designed quartz dishes have been coated with different types of UNCD films and cells were subsequently seeded on those films. Results showed the cells' growth on UNCD-coated culture dishes are similar to cell culture dishes with little retardation, indicating that UNCD films have no or little inhibition on cell proliferation and are potentially appealing as substrate/scaffold materials. The mechanisms of cell adhesion on UNCD surfaces are proposed based on the experimental results. The comparisons of cell cultures on diamond-powder-seeded culture dishes and on UNCD-coated dishes with matrix-assisted laser desorption/ionization-time-of-flight mass spectroscopy (MALDI-TOF MS) and X-ray photoelectron spectroscopy (XPS) analyses provided valuable data to support the mechanisms proposed to explain the adhesion and proliferation of cells on the surface of the UNCD platform.
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A high resolution ion mobility time-of-flight mass spectrometer with electrospray ionization source (ESI-IM-MS) was evaluated as an analytical method for rapid analysis of complex biological samples such as human blood metabolome was investigated. The hybrid instrument (IM-MS) provided an average ion mobility resolving power of ~90 and a mass resolution of ~1500 (at m/z 100). A few µL of whole blood was extracted with methanol, centrifuged and infused into the IM-MS via an electrospray ionization source. Upon IM-MS profiling of the human blood metabolome approximately 1,100 metabolite ions were detected and 300 isomeric metabolites separated in short analyses time (30 minutes). Estimated concentration of the metabolites ranged from the low micromolar to the low nanomolar level. Various classes of metabolites (amino acids, organic acids, fatty acids, carbohydrates, purines and pyrimidines etc) were found to form characteristic mobility-mass correlation curves (MMCC) that aided in metabolite identification. Peaks corresponding to various sterol derivatives, estrogen derivatives, phosphocholines, prostaglandins, and cholesterol derivatives detected in the blood extract were found to occupy characteristic two dimensional IM-MS space. Low abundance metabolite peaks that can be lost in MS random noise were resolved from noise peaks by differentiation in mobility space. In addition, the peak capacity of MS increased six fold by coupling IMS prior to MS analysis.
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Comprehensive metabolome analysis using mass spectrometry (MS) often results in a complex mass spectrum and difficult data analysis resulting from the signals of numerous small molecules in the metabolome. In addition, MS alone has difficulty measuring isobars and chiral, conformational and structural isomers. When a matrix-assisted laser desorption ionization (MALDI) source is added, the difficulty and complexity are further increased. Signal interference between analyte signals and matrix ion signals produced by MALDI in the low mass region (<1500 Da) cause detection and/or identification of metabolites difficult by MS alone. However, ion mobility spectrometry (IMS) coupled with MS (IM-MS) provides a rapid analytical tool for measuring subtle structural differences in chemicals. IMS separates gas-phase ions based on their size-to-charge ratio. This study, for the first time, reports the application of MALDI to the measurement of small molecules in a biological matrix by ion mobility-time of flight mass spectrometry (IM-TOFMS) and demonstrates the advantage of ion-signal dispersion in the second dimension. Qualitative comparisons between metabolic profiling of the Escherichia coli metabolome by MALDI-TOFMS, MALDI-IM-TOFMS and electrospray ionization (ESI)-IM-TOFMS are reported. Results demonstrate that mobility separation prior to mass analysis increases peak-capacity through added dimensionality in measurement. Mobility separation also allows detection of metabolites in the matrix-ion dominated low-mass range (m/z < 1500 Da) by separating matrix signals from non-matrix signals in mobility space.
Assuntos
Escherichia coli/metabolismo , Metabolômica/métodos , Espectrometria de Massas por Ionização por Electrospray/métodos , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz/métodos , Escherichia coli/química , Íons/química , MetabolomaRESUMO
BACKGROUND: Impaired vision, cognition, and divided attention performance predict falls. Requiring both visual and cognitive input, the ability to step accurately is necessary to safely traverse challenging terrain conditions such as uneven or slippery surfaces. We compared healthy young and older adults in the time taken to step accurately under conditions of increasing cognitive and visual demand. METHODS: Healthy Young (n = 42, mean age 21) and Older (n = 37, mean age 70) participants were required to step accurately on an instrumented walkway under conditions of increasing visual and cognitive demand. Based on the paper-and-pencil neuropsychological test, the Trail Making Test (P-TMT) A and B, participants stepped on instrumented targets with increasing sequential numbers (Walking Trail Making Test A [W-TMT A]) and increasing sequential numbers and letters (Walking Trail Making Test B [W-TMT B]), under conditions of Low as well as Normal lighting. RESULTS: W-TMT performance time increased with increased age (Older vs Young), decreased light (Low vs Normal), and increased cognitive demand (Trails B vs Trails A). W-TMT performance time was disproportionately increased in Low light and in the Older group under the highest cognitive demand (W-TMT B) conditions. Paired W-TMT A-B differences were three times higher in the Older group than in the Young group. In the Older group, the correlation between W-TMT results and P-TMT B was particularly strong (p <.001). CONCLUSIONS: The time to perform a stepping accuracy task, such as may be required to avoid environmental hazards, increases under reduced lighting and with increased cognitive demand, the latter disproportionately so in older adults.
Assuntos
Cognição/fisiologia , Desempenho Psicomotor/fisiologia , Teste de Sequência Alfanumérica , Percepção Visual/fisiologia , Caminhada/fisiologia , Adolescente , Adulto , Fatores Etários , Idoso , Idoso de 80 Anos ou mais , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Fatores de TempoRESUMO
Fall-related wrist fractures are among the most common fractures at any age. In order to learn more about the biomechanical factors influencing the impact response of the upper extremities, we studied peak hand reaction force during the bimanual arrest of a 3.4 kg ballistic pendulum moving toward the subject in the sagittal plane at shoulder height. Twenty healthy young and 20 older adults, with equal gender representation, arrested the pendulum after impact at one of three initial speeds: 1.8, 2.3, or 3.0 m/sec. Subjects were asked to employ one of three initial elbow angles: 130, 150, or 170 deg. An analysis of variance showed that hand impact force decreased significantly as impact velocity decreased (50 percent/m/s) and as elbow angle decreased (0.9 percent/degree). A two segment sagittally-symmetric biomechanical model demonstrated that two additional factors affected impact forces: hand-impactor surface stiffness and damping properties, and arm segment mass. We conclude that hand impact force can be reduced by more than 40 percent by decreasing the amount of initial elbow extension and by decreasing the velocity of the hands and arms relative to the impacting surface.