KRAS/GNAS ‐testing by highly sensitive deep targeted NGS improves the EUS ‐guided workup of suspected mucinous neoplasms of the pancreas

2021 ◽  
Author(s):  
Daniel Schmitz ◽  
Daniel Kazdal ◽  
Michael Allgäuer ◽  
Marcus Trunk ◽  
Sylke Vornhusen ◽  
...  
Keyword(s):  
Mucinous Neoplasms ◽  
Highly Sensitive ◽  
Targeted Ngs

scholarly journals Allele-Specific PCR and Next Generation Sequencing based genetic screening for Congenital Adrenal Hyperplasia in India

2021 ◽  
Author(s):  
Lavanya Ravichandran ◽  
Sophy Korula ◽  
Asha H.S. ◽  
Deny Varghese ◽  
Parthiban R ◽  
...  
Keyword(s):  
Next Generation Sequencing ◽  
Genetic Screening ◽  
Hydroxylase Deficiency ◽  
Cyp21a2 Gene ◽  
Highly Sensitive ◽  
Targeted Ngs ◽  
Allele Specific ◽  
Hotspot Mutations ◽  
21 Hydroxylase Deficiency ◽  
Generation Sequencing

Abstract Purpose Genotyping CYP21A2 gene is known to be extremely challenging and is not utilized as a first tier diagnostic tool in routine clinical practice. Also, with the advent of massive parallel sequencing technology, there is a need for investigating extended panel of genes implicated in CAH. This study aims to establish a comprehensive genetic screening strategy for five genes in CAH. Methods Allele-Specific Polymerase Chain Reaction (ASPCR) for hotspot mutations in CYP21A2 gene followed by targeted Next-Generation Sequencing (NGS) of CYP21A2, CYP11B1, CYP17A1, POR, and CYP19A1 genes were carried out to screen 72 clinically diagnosed CAH subjects from India. Results Utilizing ASPCR, 88.7% (n = 55/62) of the subjects suspected with 21 hydroxylase deficiency were positive for CYP21A2 hotspot mutations. Utilizing NGS, the ASPCR assay was found to highly sensitive and specific for screening these hotspot mutations. Additionally, through targeted NGS, six study subjects were positive for other CYP21A2 variants: one with a novel c.1274G > T, three with c.1451G > C and one with c.143A > G variant. One subject was compound heterozygous for c.955C > T/c.1042G > A variants identified using ASPCR and NGS. One subject suspected for a simple virilizing 21-hydroxylase deficiency was positive for a CYP19A1:c.1142A > T variant. CYP11B1 variants (c.1201-1G > A, c.1200 + 1delG, c.412C > T, c.1024C > T, c.1012dup, c.623G > A) were identified in all six subjects suspected for 11 beta-hydroxylase deficiency. Conclusion With an overall mutation-positivity rate of 97.2% (140/144 alleles), ASPCR followed by a multigene targeted NGS assay has shown to be highly sensitive and specific as a cost-effective and comprehensive diagnostic tool for CAH in a clinical setting.

Ovarian Dysfunction in Fetally Irradiated Beagles

1977 ◽  
Vol 35 ◽  
pp. 658-659
Author(s):  
T. M. Seed ◽  
M. H. Sanderson ◽  
D. L. Gutzeit ◽  
T. E. Fritz ◽  
D. V. Tolle ◽  
...  
Keyword(s):  
Gamma Rays ◽  
Low Dose ◽  
Long Term Effects ◽  
Ovarian Dysfunction ◽  
Dose Rates ◽  
Highly Sensitive ◽  
Microscopic Evaluation ◽  
Ovarian Pathology ◽  
Normal Offspring

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.

Fluorescence ratio imaging of dynamic intracellular ionic signals

1988 ◽  
Vol 46 ◽  
pp. 42-43
Author(s):  
R. Y. Tsien ◽  
A. Minta ◽  
M. Poenie ◽  
J.P.Y. Kao ◽  
A. Harootunian
Keyword(s):  
Binding Sites ◽  
Living Cells ◽  
Molecular Engineering ◽  
Ph Indicators ◽  
Highly Sensitive ◽  
Fluorescence Ratio Imaging ◽  
Ratio Imaging ◽  
Technical Advances ◽  
Indicator Dyes ◽  
Fluorescein Dyes

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.

Various technic for the measurement of step thickness on crystal surfaces

1978 ◽  
Vol 36 (1) ◽  
pp. 438-439
Author(s):  
C. Boulesteix ◽  
C. Colliex ◽  
C. Mory ◽  
B. Pardo ◽  
D. Renard
Keyword(s):  
Critical Thickness ◽  
Symmetric Case ◽  
Transmitted Intensity ◽  
Crystal Surfaces ◽  
Bright Field ◽  
Excited Wave ◽  
Highly Sensitive ◽  
Contrast Mechanisms ◽  
Step Thickness ◽  
Thickness Variations

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.

Estimation of the noise in TEM image recorded on “imaging plate”

1987 ◽  
Vol 45 ◽  
pp. 748-749
Author(s):  
T. Oikawa ◽  
N. Mori ◽  
T. Katoh ◽  
Y. Harada ◽  
J. Miyahara ◽  
...  
Keyword(s):  
Low Dose ◽  
Quantum Noise ◽  
Laser Light ◽  
Imaging Plate ◽  
Total System ◽  
Electron Dose ◽  
X Ray ◽  
Image Recording ◽  
Recording Material ◽  
Highly Sensitive

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.

High-sensitivity detection of gold labels by high-angle dark-field STEM imaging

1990 ◽  
Vol 48 (3) ◽  
pp. 892-893 ◽  
Author(s):  
Max T. Otten
Keyword(s):  
High Sensitivity ◽  
Dark Field ◽  
Bright Field ◽  
Backscattered Electron Imaging ◽  
Backscattered Electrons ◽  
Average Atomic Number ◽  
Highly Sensitive ◽  
Backscattered Electron ◽  
Electron Imaging ◽  
High Sensitivity Detection

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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