scholarly journals Physical characterization of a low-charge glycoform of the MUC5B mucin comprising the gel-phase of an asthmatic respiratory mucous plug

1999 ◽  
Vol 338 (2) ◽  
pp. 507-513 ◽  
Author(s):  
John K. SHEEHAN ◽  
Marjorie HOWARD ◽  
Paul S. RICHARDSON ◽  
Toby LONGWILL ◽  
David J. THORNTON

We have previously noted that sequential extraction of an asthmatic mucous exudate with 6 M guanidinium chloride yielded a fraction of the mucins that were most resistant to solubilization and of high Mr [Sheehan, Richardson, Fung, Howard and Thornton (1995) Am. J. Respir. Cell Mol. Biol. 13, 748–756]. Here we show that this mucin fraction is dominated (at least 96% of the total) by the low-charge glycoform of the MUC5B gene product. Seen in the electron microscope the mucins appeared mainly as compact ‘island ’ structures composed of linear threads often emanating from globular ‘nodes ’ rather than the discrete linear threads more typical of mucins that we have previously described. The effect of reducing agents was as expected for other gel-forming mucins, i.e. reduced subunits or monomers of Mr 3×106) were produced within 15 min of treatment. Kinetic experiments on the cleavage of the intact mucins with the proteinase trypsin indicated two clear regimes of fragmentation. An initial rapid cleavage generated mucins ranging from Mr = 4×106 to 30×106 that in the electron microscope appeared as polydisperse threads (500–3000 nm in length), similar to normal and other respiratory mucins that we have previously characterized. A subsequent slower fragmentation over many hours yielded a major fragment of Mr 3×106 and length 200–600 nm, very similar in size and Mr to the subunits obtained by reduction. The results suggest that the MUC5B mucin is assembled, first into polydisperse linear threads, which are then linked together via a protein-mediated process. This might involve part of the mucin polypeptide or an as yet unidentified protein(s). The high proteinase susceptibility of the linkage suggests that it might be a point of control for mucin size and thus mucus rheology.

1999 ◽  
Vol 338 (2) ◽  
pp. 507 ◽  
Author(s):  
John K. SHEEHAN ◽  
Marjorie HOWARD ◽  
Paul S. RICHARDSON ◽  
Toby LONGWILL ◽  
David J. THORNTON

Author(s):  
R. F. Schneidmiller ◽  
W. F. Thrower ◽  
C. Ang

Solid state materials in the form of thin films have found increasing structural and electronic applications. Among the multitude of thin film deposition techniques, the radio frequency induced plasma sputtering has gained considerable utilization in recent years through advances in equipment design and process improvement, as well as the discovery of the versatility of the process to control film properties. In our laboratory we have used the scanning electron microscope extensively in the direct and indirect characterization of sputtered films for correlation with their physical and electrical properties.Scanning electron microscopy is a powerful tool for the examination of surfaces of solids and for the failure analysis of structural components and microelectronic devices.


Author(s):  
T. C. Tisone ◽  
S. Lau

In a study of the properties of a Ta-Au metallization system for thin film technology application, the interdiffusion between Ta(bcc)-Au, βTa-Au and Ta2M-Au films was studied. Considered here is a discussion of the use of the transmission electron microscope(TEM) in the identification of phases formed and characterization of the film microstructures before and after annealing.The films were deposited by sputtering onto silicon wafers with 5000 Å of thermally grown oxide. The film thicknesses were 2000 Å of Ta and 2000 Å of Au. Samples for TEM observation were prepared by ultrasonically cutting 3mm disks from the wafers. The disks were first chemically etched from the silicon side using a HNO3 :HF(19:5) solution followed by ion milling to perforation of the Au side.


Author(s):  
Ruchama Baum ◽  
J.T. Seto

The ribonucleic acid (RNA) of paramyxoviruses has been characterized by biochemical and physiochemical methods. However, paramyxovirus RNA molecules have not been studied by electron microscopy. The molecular weights of these single-stranded viral RNA molecules are not known as yet. Since electron microscopy has been found to be useful for the characterization of single-stranded RNA, this investigation was initiated to examine the morphology and length measurements of paramyxovirus RNA's.Sendai virus Z strain and Newcastle disease virus (NDV), Milano strain, were used. For these studies it was necessary to develop a method of extracting RNA molecules from purified virus particles. Highly purified Sendai virus was treated with pronase (300 μg/ml) at 37°C for 30 minutes and the RNA extracted by the sodium dodecyl sulfate (SDS)-phenol procedure.


Author(s):  
Robert C. Rau

Previous work has shown that post-irradiation annealing, at temperatures near 1100°C, produces resolvable dislocation loops in tungsten irradiated to fast (E > 1 MeV) neutron fluences of about 4 x 1019 n/cm2 or greater. To crystallographically characterize these loops, tilting experiments were carried out in the electron microscope on a polycrystalline specimen which had been irradiated to 1.5 × 1021 n/cm2 at reactor ambient temperature (∼ 70°C), and subseouently annealed for 315 hours at 1100°C. This treatment produced large loops averaging 1000 Å in diameter, as shown in the micrographs of Fig. 1. The orientation of this grain was near (001), and tilting was carried out about axes near [100], [10] and [110].


Author(s):  
George Guthrie ◽  
David Veblen

The nature of a geologic fluid can often be inferred from fluid-filled cavities (generally <100 μm in size) that are trapped during the growth of a mineral. A variety of techniques enables the fluids and daughter crystals (any solid precipitated from the trapped fluid) to be identified from cavities greater than a few micrometers. Many minerals, however, contain fluid inclusions smaller than a micrometer. Though inclusions this small are difficult or impossible to study by conventional techniques, they are ideally suited for study by analytical/ transmission electron microscopy (A/TEM) and electron diffraction. We have used this technique to study fluid inclusions and daughter crystals in diamond and feldspar.Inclusion-rich samples of diamond and feldspar were ion-thinned to electron transparency and examined with a Philips 420T electron microscope (120 keV) equipped with an EDAX beryllium-windowed energy dispersive spectrometer. Thin edges of the sample were perforated in areas that appeared in light microscopy to be populated densely with inclusions. In a few cases, the perforations were bound polygonal sides to which crystals (structurally and compositionally different from the host mineral) were attached (Figure 1).


Author(s):  
E. Hendarto ◽  
S.L. Toh ◽  
J. Sudijono ◽  
P.K. Tan ◽  
H. Tan ◽  
...  

Abstract The scanning electron microscope (SEM) based nanoprobing technique has established itself as an indispensable failure analysis (FA) technique as technology nodes continue to shrink according to Moore's Law. Although it has its share of disadvantages, SEM-based nanoprobing is often preferred because of its advantages over other FA techniques such as focused ion beam in fault isolation. This paper presents the effectiveness of the nanoprobing technique in isolating nanoscale defects in three different cases in sub-100 nm devices: soft-fail defect caused by asymmetrical nickel silicide (NiSi) formation, hard-fail defect caused by abnormal NiSi formation leading to contact-poly short, and isolation of resistive contact in a large electrical test structure. Results suggest that the SEM based nanoprobing technique is particularly useful in identifying causes of soft-fails and plays a very important role in investigating the cause of hard-fails and improving device yield.


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