Composition of nasal airway surface liquid in cystic fibrosis and other airway diseases determined by X-ray microanalysis

The ionic composition of the airway surface liquid (ASL) in healthy individuals and in patients with cystic fibrosis (CF) has been debated. Ion transport properties of the upper airway epithelium are similar to those of the lower airways and it is easier to collect nasal ASL from the nose. ASL was collected with ion exchange beads, and the elemental composition of nasal fluid was determined by X-ray microanalysis in healthy subjects, CF patients, CF heterozygotes, patients with rhinitis, and with primary ciliary dyskinesia (PCD). In healthy subjects, the ionic concentrations were approximately isotonic. In CF patients, CF heterozygotes, rhinitis, and PCD patients, [Na] and [Cl] were significantly higher compared when compared with those in controls. [K] was significantly higher in CF and PCD patients compared with that in controls. Severely affected CF patients had higher ionic concentrations in their nasal ASL than in patients with mild or moderate symptoms. Female CF patients had higher levels of Na, Cl, and K than male patients. As higher salt concentrations in the ASL are also found in other patients with airway diseases involving chronic inflammation, it appears likely that inflammation-induced epithelial damage is important in determining the ionic composition of the ASL.     

Assessment of chloride secretion in human nasal epithelial cells by X-ray microanalysis

The genetic disease cystic fibrosis (CF) is due to defective epithelial chloride transport. Different treatments have been proposed that could restore chloride transport in CF patients. A new method is proposed for measuring the chloride secretion in easily accessible epithelial cells.
Fresh nasal epithelial cells were obtained by nasal brushing and made to attach to titanium grids for electron microscopy. Chloride efflux through the cystic fibrosis transmembrane regulator channel was stimulated by 20 µ m forskolin and 100 µ m isobutyl-methylxanthine (IBMX), in standard Ringer's solution (SR). Chloride efflux through the calcium-regulated channel was stimulated by 200 µ m adenosine triphosphate (ATP) in SR. The cells were rinsed after the exposure, in order to remove the experimental medium, frozen and freeze-dried. The elemental composition of the cells was determined by X-ray microanalysis.
Rinsing with distilled water or ammonium acetate appeared to cause damage to the cells, whereas rinsing with isotonic mannitol preserved the ionic composition. Stimulation of cells from healthy controls with forskolin and IBMX in a chloride-containing medium caused a significant (28 ± 6%) decrease in chloride concentration, which is indicative of net chloride efflux. In similar conditions, stimulation with ATP induced a 29 ± 5% decrease in the chloride concentration.
Stimulation of cells from CF patients with forskolin and IBMX in a chloride-containing medium caused no significant change in the intracellular chloride concentration, whereas ATP stimulation induced a response similar to that obtained in cells from healthy controls.
It is concluded that X-ray microanalysis of nasal epithelial cells may be used to determine chloride secretion in CF patients in an easily accessible cell type.     

A home-based pulmonary function monitor for cystic fibrosis

As care for the chronically ill makes increasing demands upon our medical system, cost-effective methods for addressing those demands are being sought. The use of self-monitoring coupled with telecommunication of results to the health-care provider can be expected to provide a partial solution in the appropriate clinical setting. An electronic spirometer with telecommunication ability for use by cystic fibrosis patients in the home has been developed.     

Methods for determining the composition of nasal fluid by X-ray microanalysis

The nasal fluid is an easily accessible form of airway surface liquid. The objective of this study was to find a technically easy and reproducible method for sampling and analysis of this fluid. In a pilot study, several methods to carry out X-ray microanalysis of sub-microliter droplets were compared. Acceptable results were obtained with several of these methods (pipeting on filter paper or analysis of frozen-hydrated droplets at low temperature). Nasal fluid was collected from the inferior turbinate with a micropipette after occlusion of a nostril for 5-10 minutes. Ion concentrations in nasal fluid from six control subjects were (in mM, mean +/- standard error): sodium (Na) 127 +/- 6, chloride (Cl) 140 +/- 7, potassium (K) 27 +/- 3, and calcium (Ca) 5 +/- 1. This sampling method proved difficult to apply to cystic fibrosis (CF) patients because of the viscous quality of their nasal secretion. Therefore, an alternative method was devised. Sephadex G-25, ion exchange beads were mounted on double-sided tape, which was stuck on a filter paper as support. The filter paper was applied for 10 minutes to the nostril of a subject, and kept loosely in place. During the exposure period, the nasal fluid equilibrates with the beads. After removal of the filter paper with the beads from the nostril, the beads were rinsed with a hydrophobic volatile silicone oil to remove excess nasal fluid, dried, and analyzed. This method of collection is not cumbersome for the subject and gives results similar to those obtained by the direct collection method: Na 142 +/- 28 mM, Cl 150 +/- 36 mM, K 43 +/- 10 mM (mean and standard error of four determinations). Small differences between the filter method and the bead method can be explained by the fact that the filter method measured total nasal fluid, whereas the bead method measures predominantly the fluid component. Subjects suffering from mild respiratory illness or rhinitis had higher values for Na, K, and Cl in their nasal fluid.     

Paired helical filaments (PHFs) are a family of single filament structures with a common helical turn period: negatively stained PHF imaged by TEM and measured before and after sonication, deglycosylation, and dephosphorylation

Isolated paired helical filaments (PHFs) were visualized on glutaraldehyde vapor-treated thin approximately 10-nm thick indirect carbon films using high-resolution transmission electron microscopy (TEM) and the negative stain, phosphotungstate acid (PTA) at near neutral pH of 6.8. PHF preparations were prepared with and without 1 minute of sonication. These same PHF were also deglycosylated with endoglycosidase F/N-glycosidase F for 1 hour or the PHF were dephosphorylated with PP-2A for 1 hour. The negatively stained PHF filaments were quantitatively studied by measuring their wide regions (W) their thin regions (T) and their helical turn period (L) and these separate parameters were averaged for each filament. In the unsonicated PHF preparation there were PHF, cylindrical filaments with periodic thin regions (CF-PT), cylindrical filaments (CF), as well as 2.0-nm tau polymer-like filaments. The CF-PT were characterized by W, T, and L measurements and the CF were characterized by diameter measurements. The paired helical filament model proposed by Kidd (1963, Nature 197:192-193) of two approximately 10 nm filaments twisting around each other every approximately 80 nm with a thin region of 10 nm and a wide region of 25 nm does not correspond to the PHF structures found. None of the PHF we observed were composed of a pair of filaments and all of the PHF appear to be a single filament. The wide regions ranged from 12.5-27 nm and the thin regions ranged from 4.5-12.3 nm. The helical turn periods ranged from 76-85 nm and were generally about 80 nm. Only the helical turn period of approximately 80 nm was a common property of the whole family of PHF structures. The CF-PT appear to be a PHF precursor filament. Deglycosylation of the PHF and CF-PT reduced their sizes by 0.5-0.6 nm and 0.7-1.0 nm, respectively, and the right-hand helicity of the PHF was lost after deglycosylation. Dephosphorylation with PP-2A reduced the PHF wide regions by 6.0 nm and the thin regions by 2.6 nm.     

Effect of processing methods for transmission electron microscopy on corneal collagen fibrils diameter and spacing

Introduction: The corneal tissue was processed in fixatives and embedded in resin for transmission electron microscopy to observe the ultrastructure of the collagen fibrils (CFs). The effect of these processing methods on the CF diameter and the interfibrillar spacing was studied. Methods: Four normal human corneal buttons were used for this study. A part of each cornea was fixed in 2.5% glutaraldehyde containing cuprolinic blue in sodium acetate buffer and embedded in spurr's resin (SpurrCB). A second part of each cornea was fixed in 2.5% glutaraldehyde + osmium tetroxide and embedded spurr's resin (SpurrOsm). The third part of each cornea was fixed in paraformaldehyde (4%) and embedded in LR White at 4°C (LRWhite). Ultrathin sections were stained with uranyl acetate and lead citrate. Results: In the tissue, fixed in SpurrCB, the diameter was 38.4 ± 5.9 nm and spacing between CF was 52.5 ± 5.3 nm. In the tissue fixed in SpurrOsm, the diameter was 28.37 ± 5.84 nm and spacing between CF was 45 ± 4.57 nm. In the tissue fixed in LR White, the CF diameter was 24 ± 2.3 nm and spacing between CF was 39.0 ± 4.2 nm. The diameters and interfibrillar spacing of the tissue processed by SpurrCB, SpurrOsm, and LRWhite were significantly different (P < 0.001) from one another. Conclusion: Our study shows that there is a variation in the CF diameter and spacing depending on the method of fixation and embedding resins used. This needs to be considered when comparative studies using different methods are done. Microsc. Res. Tech. 2012. © 2012 Wiley Periodicals, Inc.     

Solid particle erosion of unidirectional carbon fibre reinforced polyetheretherketone composites

The solid particle erosion behaviour of unidirectional carbon fibre (CF) reinforced polyetheretherketone (PEEK) composites has been characterised. The erosion rates of these composites have been evaluated at different impingement angles (15–90°) and at three different fibre orientations (0, 45, and 90°). The particles used for the erosion measurements were steel balls with diameter of 300–500 μm and impact velocities of 45 and 85 m/s. The unidirectional CF reinforced PEEK composites showed semi-ductile erosion behaviour, with maximum erosion rate at 60° impingement angle. The fibre orientations had a significant influence on erosion rate. The morphology of eroded surfaces was examined by using scanning electron microscopy (SEM). Possible erosion mechanisms are discussed.     

Elemental analysis of cemented carbides by calibration-free portable laser-induced breakdown spectroscopy

The process of cemented carbides manufacturing requires rapid and field elemental analytical techniques to control and evaluate the properties of products. Calibration-free laser-induced breakdown spectroscopy (CF-LIBS) is such a potential elemental analytical technique. In this work, a portable LIBS instrument combined with a CF method was developed for the analysis of cemented carbides. Three batches of cemented compact carbides without reference samples were analyzed. Qualitative and quantitative analysis of the samples were achieved by using the portable LIBS instrument combined with CF method. To validate the analysis results, X-ray fluorescence spectrometry (XRF) was used to analyze the samples as well. The results of CF-LIBS agreed well with the results of XRF, with relative errors between ?29.53 and 24.70%. The results demonstrated that the portable LIBS instrument combined with CF method was capable for direct and rapid analysis without any need of standard measurements. Notably, with the portable LIBS instrument combined with CF method, acceptable accuracy could be obtained, which is promising for practical field applications.     

Effect of rare earths on mechanical and tribological properties of carbon fibers reinforced PTFE composite

The effect of a rare earth (RE) surface treatment on the mechanical and tribological properties of carbon fiber (CF) reinforced polytetrafluoroethylene (PTFE) composites was experimentally investigated. The tensile properties of the CF reinforced PTFE (CF/PTFE) composites treated with air oxidation and RE modifier were superior to those of untreated CF/PTFE composites, while RE treatment was most effective in promoting the tensile strength and strain at break of the CF/PTFE composite. The bending strength of the RE treated CF/PTFE composite was improved by about 16% compared with that of untreated composites, while 2% improvement was achieved by air oxidation. Under oil-lubricated conditions, RE treatment was more effective than air oxidation to reduce the friction coefficient and wear of PTFE composite. RE treatment effectively improved the interfacial adhesion between CF and PTFE. The strong interfacial coupling of the composite made CF not easy to detach from the PTFE matrix, and prevented the rubbing-off of PTFE, accordingly improved the friction and wear properties of the composite.     

Ion mixing at 20 keV: a comparison of the effects of Ga+, Ar+ and CF4+ ion irradiation

Medium-energy (some tens of keV) ion irradiation is frequently used in various technologies. It is well known that during this irradiation serious alterations are introduced to the material, changing its structure, composition, etc. While there are studies on the amorphization, no results have been reported on the medium-energy ion beam-induced mixing, however. In this work, we present Auger electron spectroscopy (AES) depth profiling measurements of Si/Cr multilayer samples, which were irradiated by various ions (Ga+, Ar+, CF4+) of 20 keV applying angles of incidence of 5 degrees (Ga+), 65 degrees (Ga+) and 75 degrees (Ar+, CF4+). The ion beam-induced mixing at the Si/Cr interface (the broadening of the interface) was measured as a function of the removed layer thickness. The weakest and strongest ion mixing (for a given removed layer thickness) were found for CF4+ and Ga+ 5 degrees irradiations, respectively. In the case of Ga+ irradiation, the larger the angle of incidence the weaker the ion mixing. The extent of mixing does not correlate with the corresponding projected range. Comparison of the experimentally measured ion mixed profiles with those given by dynamic TRIM simulations gave poor agreement for Ar+ and fails for Ga+ irradiations, respectively.     

Visualization of single proteins from stripped native cell membranes: A protocol for high‐resolution atomic force microscopy

Atomic force microscopy (AFM) proved to be able to obtain high‐resolution three‐dimensional images of single‐membrane proteins, isolated, crystallized, or included in reconstructed model membranes. The extension of this technique to native systems, such as the protein immersed in a cell membrane, needs a careful manipulation of the biological sample to meet the experimental constraints for high‐resolution AFM imaging. In this article, a general protocol for sample preparation is presented, based on the mechanical stretch of the cell membrane. The effectiveness for AFM imaging has been tested on the basis of an integrated optical and AFM approach and the proposed method has been applied to cells expressing cystic fibrosis transmembrane conductance regulator, a channel involved in cystic fibrosis, showing the possibility to identify and analyze single proteins in the plasma membrane. Microsc. Res. Tech. 76:723–732, 2013. © 2013 Wiley Periodicals, Inc.     

Stiffness Analysis of Corrugated Flexure Beam Used in Compliant Mechanisms

Conventional flexible joints generally have limited range of motion and high stress concentration. To overcome these shortcomings, corrugated flexure beam(CF beam) is designed because of its large flex...     

Estimation of the daily global solar radiation; Nepal experience

This work focuses on estimation of global solar radiation and, in particular, it explores the effect of precipitation and temperature on solar radiation profile of Kathmandu (Nepal). An accurate knowledge of solar radiation distribution in each particular geographical location is crucial for the promotion of solar energy technology. The best way of knowing the amount of global solar radiation is to install quality instruments at many locations in the given region. This requires their day to day maintenance, recording and calibration, which is very costly in developing countries like Nepal. Thus, the alternative approach is to correlate meteorological measurements with appropriate models and investigate the key parameters. For the research activities the RadEst program has been used. It includes, for evaluating the daily global solar radiation values at a given latitudes, four models which estimate the atmospheric transmissivity by measurements of daily temperature range and precipitations. The model parameters are fitted in 2 years data by iterative procedures. The values obtained by these models are, then, compared with measured radiation data. The paper reports graphical and statistical evaluations suggesting that among the four models, the Modular DCBB is the best model for Kathmandu area. This result is useful for designing solar panels able to maximize the harvesting of solar energy and to reduce the chronic shortage of hydrocarbon fuel that Nepal imports in a significant amount each year.     

A simple approach to standardized spinal radiographs

Present methods of obtaining spine radiographs for the clinical assessment of scoliosis are inadequate. Routine quantitative measurements made on anterior-posterior (A-P) and lateral radiographs describing the spinal deformities are grossly inaccurate since they are two-dimensional measurements of three-dimensional deformities. The approach described in this paper uses a simple device, the "Throne," which ensures reproducible positioning of patients for sitting spine radiographs as well as exact orthogonality of A-P and lateral film exposures. These two views of the spine form the data in digitized form for input into a computer program which generates the 3-D geometry of the line passing through the centers of each vertebral body from L5 to T1. Parameters describing the 3-D deformities of the spine are calculated automatically and converted to printed form for detailed examination and to plotted form for clinical use. Approximately 80 postambulatory Duchenne muscular dystrophy patients are being followed using this method. Of these, 20 have been followed for periods of 2 to 3 years. Costs of the method are minimal. The "Throne" can be built for approximately $50 of materials and 30 hours of labor. Each set of A-P and lateral radiographs can be processed in less than 5 minutes, including digitizing time at a computer cost of about 50 cents. The computer program may be handled by a minicomputer.     

CT定位下微创软通道术在高龄20~30ml幕上血肿中的应用

目的:探讨CT定位下微创软通道术在高龄20~30ml幕上血肿中的应用效果。方法:选取2018年6月~2020年9月本院收治的60例高龄20~30mL幕上血肿患者,随机分为微创组及保守组,各30例。微创组采取CT定位下微创软通道手术治疗,保守组采取保守疗法。对比治疗前及出院时的美国国立卫生研究院卒中量表(NIHSS)评分及临床效果,并对比治疗前、治疗7d、治疗1个月的日常生活能力(Barthel指数评估),另比较治疗后3个月的预后情况[格拉斯哥预后量表(GOS)评价]及并发症发生情况。结果:两组出院时NIHSS评分均较治疗前降低,微创组出院时NIHSS评分低于保守组,差异均具有统计学意义(P<0.05);两组临床效果等级分布比较有统计学差异(P<0.05),微创组总有效率高于保守组,差异有统计学意义(P<0.05);Barthel指数评分在组间、时间、交互方面具有统计学差异(P<0.05),两组治疗7d、治疗后1个月Barthel指数均较治疗前升高,两组治疗1个月均高于治疗7d,微创组治疗7d、治疗后1个月均高于微创组,差异均具有统计学意义(P<0.05);两组治疗后3个月的GOS评分等级分布比较有统计学差异(P<0.05),微创组预后良好率高于保守组,差异有统计学意义(P<0.05);微创组总并发症发生率与保守组比较差异无统计学意义(P>0.05)。结论:CT定位下微创软通道手术在高龄20~30mL幕上血肿患者治疗中的临床效果、神经功能恢复、日常生活能力改善方面均优于保守疗法,且预后改善更为明显,安全性较好。     

双羟氟轻松有关物质测定方法研究

目的:用HPLC法测定双羟氟轻松的有关物质。方法:Agilent TC-C_(18)柱为分离柱,0.05%的H_3PO_4-甲醇(43:57)为流动相,流速为1.0mL/min,检测波长为241nm。结果:双羟氟轻松的主峰和杂质峰能有效地分离,重现性良好。双羟氟轻松检测浓度在1.501～15.010mg/mL范围内与峰面积呈良好线性关系,r=0.9999(n=5);检测限分别为:双羟氟轻松为0.18mg/mL、中间体CF11为0.74mg/mL、CF12为0.25mg/mL、CF13为0.20mg/mL。结论:本方法准确、简便、快速、专属性强、重现性好,能很好地控制双羟氟轻松的质量。     

碳纤维增强尼龙1010复合材料的摩擦磨损性能及磨损机理研究

以注塑成型法制备了尼龙1010及碳纤维(CF)增强尼龙1010复合材料，研究了CF含量和载荷对材料摩擦学性能和磨损机制的影响。结果表明，CF的加入可显著改善尼龙的摩擦学性能，以体积分数为20％的CF增强尼龙1010复合材料的耐磨性能最好。较低的CF含量下复合材料磨损表面主要受到对偶钢环上微凸峰的切削和犁沟作用，较高载荷时发生了热疲劳剥层磨损；随着CF含量增加，复合材料表面在较高载荷时产生明显疲劳断裂，并使对偶钢环产生较剧烈磨损。     

不同方法表面改性碳纤维增强热塑性聚酰亚胺复合材料的摩擦磨损性能

采用硝酸氧化改性和涂层复合改性法分别对碳纤维(CF)进行了表面处理,并制备了CF增强热塑性聚酰亚胺(TPI)复合材料;对CF的表面形貌进行了观察,研究了表面改性方法对复合材料摩擦磨损性能的影响,并利用扫描电子显微镜观察了磨损表面形貌。结果表明:硝酸氧化改性增大了CF的表面粗糙度,随处理时间的延长粗糙度增大;经涂层复合改性后,CF表面包覆了一层聚酰亚胺(PI),保护了CF并提高了其与基体界面的结合强度;经表面改性后的CF增强TPI复合材料的摩擦磨损性能均得到提高,以涂层复合改性的效果最好;硝酸氧化改性后的CF在摩擦过程中易断裂,复合材料的磨损形貌以磨粒磨损为主,而涂层复合改性后的CF断裂得到抑制,与基体结合更为牢固,磨损表面较为平整。     

Measurement of laxity in the anterior cruciate ligament-deficient knee: a comparison of three different methods in vitro

The aim of this study was to compare in-vitro measurements of anteroposterior laxity in the anterior cruciate ligament (ACL)-deficient knee using three different methods: an Instron materials-testing machine, then a KT-2000 arthrometer, and finally by Roentgen stereophotogrammetric analysis (RSA). Eight ACL-deficient human cadaver knees were used. Total displacement was measured between 90 N anterior and 90 N posterior tibiofemoral drawer forces at both 20 degrees and 90 degrees knee flexion. Laxity ranged from 11.5 to 27.6 mm at 20 degrees and from 8.7 to 23.9 mm at 90degrees. A statistically significant difference was not found between the mean RSA and KT-2000 measurements. However, the mean Instron measurements of laxity were significantly (3-4 mm) higher than both RSA and KT-2000 measurements. The clinical methods of RSA and the KT-2000 measurements agreed well but appeared to underestimate tibiofemoral anteroposterior laxity compared with the materials-testing machine. These findings may be helpful in the future comparison of different studies.