RAMA casein zymography: Time-saving and highly sensitive casein zymography for MMP7 and trypsin

2016 ◽  
Vol 37 (22) ◽  
pp. 2959-2962 ◽  
Author(s):  
Hidetaro Yasumitsu ◽  
Yasuhiro Ozeki ◽  
Robert A. Kanaly
2021 ◽  
Vol 9 (4) ◽  
pp. 765
Author(s):  
Janika Wolff ◽  
Martin Beer ◽  
Bernd Hoffmann

Outbreaks of the three capripox virus species, namely lumpy skin disease virus, sheeppox virus, and goatpox virus, severely affect animal health and both national and international economies. Therefore, the World Organization for Animal Health (OIE) classified them as notifiable diseases. Until now, discrimination of capripox virus species was possible by using different conventional PCR protocols. However, more sophisticated probe-based real-time qPCR systems addressing this issue are, to our knowledge, still missing. In the present study, we developed several duplex qPCR assays consisting of different types of fluorescence-labelled probes that are highly sensitive and show a high analytical specificity. Finally, our assays were combined with already published diagnostic methods to a diagnostic workflow that enables time-saving, reliable, and robust detection, differentiation, and characterization of capripox virus isolates.


Author(s):  
N Aydin ◽  
F Chardonnens ◽  
M Rotach

AbstractBecause many physicochemical properties of tobacco are highly sensitive to its moisture content, the determination of water level is an important parameter for tobacco characterization. A headspace volumetric Karl Fischer titration (HS-V-KFT) method is presented for the quantification of water content in different finished tobacco materials. The parameters affecting the extraction of water from the tobacco materials were the sample size and the oven temperature which have been optimized. The extraction of water from the samples was achieved within a reasonable time (<25 min) with a sample size of 200 mg and an optimum temperature of between 90 °C and 100 °C. The results of the water determination by HS-V-KFT at the optimized parameters were in good agreement with those obtained by standard volumetric Karl Fischer titration. HS-V-KFT showed very good repeatability (RSDr 0.9%) and intermediate precision (RSDiR 1.1%). With respect to a considerable time saving, solvent consumption reduction, precision and accuracy, HS-V-KFT can therefore be suggested as the method of choice to determine water amount in finished tobacco products.


2019 ◽  
Vol 19 (11) ◽  
pp. 7229-7235
Author(s):  
Anila Arshad ◽  
Raheel Akram ◽  
Shahid Iqbal Rana ◽  
Ghulam Hussain ◽  
Sami Ullah Dar ◽  
...  

A single step fast track method is proposed to determine melamine based on strong hydrogen bonding between exocyclic amine group in melamine moiety and hydroxyl group of tannic acid (TA); acting as reducing and functionalizing agent for synthesis of gold nanoparticles (Au NPs). The proposed strategy is facile and highly time saving; as the detection of melamine with labelfree gold nanoparticles was accomplished within five minutes. The colorimetric probe exhibits high detection limit of 8 nM for melamine determination in raw milk, which is very low as compared to other capping ligands employed for Au NPs such as borohydride, citrate and 3,5-dihydroxybenzoic acid. The results show a highly sensitive linear range (0.03–2.5 μM) between absorption ratio and concentration of melamine with a correlation coefficient of 0.991. Proposed methodology is proven effective for on-site screening without using costly instrumentation.


Author(s):  
T. M. Seed ◽  
M. H. Sanderson ◽  
D. L. Gutzeit ◽  
T. E. Fritz ◽  
D. V. Tolle ◽  
...  

The developing mammalian fetus is thought to be highly sensitive to ionizing radiation. However, dose, dose-rate relationships are not well established, especially the long term effects of protracted, low-dose exposure. A previous report (1) has indicated that bred beagle bitches exposed to daily doses of 5 to 35 R 60Co gamma rays throughout gestation can produce viable, seemingly normal offspring. Puppies irradiated in utero are distinguishable from controls only by their smaller size, dental abnormalities, and, in adulthood, by their inability to bear young.We report here our preliminary microscopic evaluation of ovarian pathology in young pups continuously irradiated throughout gestation at daily (22 h/day) dose rates of either 0.4, 1.0, 2.5, or 5.0 R/day of gamma rays from an attenuated 60Co source. Pups from non-irradiated bitches served as controls. Experimental animals were evaluated clinically and hematologically (control + 5.0 R/day pups) at regular intervals.


Author(s):  
R. Y. Tsien ◽  
A. Minta ◽  
M. Poenie ◽  
J.P.Y. Kao ◽  
A. Harootunian

Recent technical advances now enable the continuous imaging of important ionic signals inside individual living cells with micron spatial resolution and subsecond time resolution. This methodology relies on the molecular engineering of indicator dyes whose fluorescence is strong and highly sensitive to ions such as Ca2+, H+, or Na+, or Mg2+. The Ca2+ indicators, exemplified by fura-2 and indo-1, derive their high affinity (Kd near 200 nM) and selectivity for Ca2+ to a versatile tetracarboxylate binding site3 modeled on and isosteric with the well known chelator EGTA. The most commonly used pH indicators are fluorescein dyes (such as BCECF) modified to adjust their pKa's and improve their retention inside cells. Na+ indicators are crown ethers with cavity sizes chosen to select Na+ over K+: Mg2+ indicators use tricarboxylate binding sites truncated from those of the Ca2+ chelators, resulting in a more compact arrangement of carboxylates to suit the smaller ion.


Author(s):  
C. Boulesteix ◽  
C. Colliex ◽  
C. Mory ◽  
B. Pardo ◽  
D. Renard

Contrast mechanisms, which are responsible of the various types of image formation, are generally thickness dependant. In the following, two imaging modes in the 100 kV CTEM are described : they are highly sensitive to thickness variations and can be used for quantitative estimations of step heights.Detailed calculations (1) of the bright-field intensity have been carried out in the 3 (or 2N+l)-beam symmetric case. They show that in given conditions, the two important symmetric Bloch waves interfere most strongly at a critical thickness for which they have equal emergent amplitudes (the more excited wave at the entrance surface is also the more absorbed). The transmitted intensity I for a Nd2O3 specimen has been calculated as a function of thickness t. The capacity of the method to detect a step and measure its height can be more clearly deduced from a plot of dl/Idt as shown in fig. 1.


Author(s):  
B. Carragher ◽  
M. Whittaker

Techniques for three-dimensional reconstruction of macromolecular complexes from electron micrographs have been successfully used for many years. These include methods which take advantage of the natural symmetry properties of the structure (for example helical or icosahedral) as well as those that use single axis or other tilting geometries to reconstruct from a set of projection images. These techniques have traditionally relied on a very experienced operator to manually perform the often numerous and time consuming steps required to obtain the final reconstruction. While the guidance and oversight of an experienced and critical operator will always be an essential component of these techniques, recent advances in computer technology, microprocessor controlled microscopes and the availability of high quality CCD cameras have provided the means to automate many of the individual steps.During the acquisition of data automation provides benefits not only in terms of convenience and time saving but also in circumstances where manual procedures limit the quality of the final reconstruction.


Author(s):  
T. Oikawa ◽  
N. Mori ◽  
T. Katoh ◽  
Y. Harada ◽  
J. Miyahara ◽  
...  

The “Imaging Plate”(IP) is a highly sensitive image recording plate for X-ray radiography. It has been ascertained that the IP has superior properties and high practicability as an image recording material in a TEM. The sensitivity, one of the properties, is about 3 orders higher than that of conventional photo film. The IP is expected to be applied to low dose techniques. In this paper, an estimation of the quantum noise on the TEM image which appears in case of low electron dose on the IP is reported.In this experiment, the JEM-2000FX TEM and an IP having the same size as photo film were used.Figure 1 shows the schematic diagram of the total system including the TEM used in this experiment. In the reader, He-Ne laser light is scanned across the IP, then blue light is emitted from the IP.


Author(s):  
Max T. Otten

Labelling of antibodies with small gold probes is a highly sensitive technique for detecting specific molecules in biological tissue. Larger gold probes are usually well visible in TEM or STEM Bright-Field images of unstained specimens. In stained specimens, however, the contrast of the stain is frequently the same as that of the gold labels, making it virtually impossible to identify the labels, especially when smaller gold labels are used to increase the sensitivity of the immunolabelling technique. TEM or STEM Dark-Field images fare no better (Figs. 1a and 2a), again because of the absence of a clear contrast difference between gold labels and stain.Potentially much more useful is backscattered-electron imaging, since this will show differences in average atomic number which are sufficiently large between the metallic gold and the stains normally used. However, for the thin specimens and at high accelerating voltages of the STEM, the yield of backscattered electrons is very small, resulting in a very weak signal. Consequently, the backscattered-electron signal is often too noisy for detecting small labels, even for large spot sizes.


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