scholarly journals EPEN-14. GENERATION OF A C11orf95-RELA FUSION TARGETING ANTIBODY AS A DIAGNOSTIC TOOL FOR SUPRATENTORIAL EPENDYMOMA

2020 ◽  
Vol 22 (Supplement_3) ◽  
pp. iii310-iii310
Author(s):  
Lisa Ruff ◽  
Denice Y Chan ◽  
Lesley Jenkinson ◽  
Stuart Haynes ◽  
Mark Austin ◽  
...  
Keyword(s):  
Diagnostic Tool ◽  
Fusion Gene ◽  
Affinity Maturation ◽  
Mouse Tissue ◽  
Great Promise ◽  
Nuclear Staining ◽  
Screening Process ◽  
Similar Treatment ◽  
Genomic Studies ◽  
Supratentorial Ependymoma

Abstract Ependymomas account for 10% of paediatric brain tumours and arise in the ventricular walls of the central nervous system. Ependymomas were previously classified as one tumour type and all patients received similar treatment. However, recent genomic studies have identified nine different molecular subgroups of the disease, including one supratentorial subtype characterized by a novel fusion gene C11ORF95-RELA. When introduced into neural stem cells, this fusion is a potent driver of tumorigenesis and its presence in patient samples has previously also been shown to negatively correlate with overall survival. Accurate diagnosis of this subgroup is currently limited to sophisticated approaches such as break-apart FISH or RNA sequencing. Here, we report the generation of a C11ORF95-RELA Fusion-specific antibody that can be used for routine immunohistochemistry (IHC). Candidate antibodies were first selected using phage display and favourable leads were subjected to affinity maturation using ribosome display after a screening process involving immunoblotting and IHC. Further IHC-based screening of affinity-matured candidates using fusion-positive and -negative mouse tissue as well as human fusion-negative ependymoma tumour tissue produced one lead antibody. The antibody detects fusion-specific nuclear staining pattern on fusion-positive tissue and does not react with fusion-negative tissues. This candidate antibody is currently being tested on human fusion-positive ependymoma tissue. This accurate diagnostic tool holds great promise to transform the management of patients with supratentorial ependymoma.

scholarly journals Infrared spectral histopathology using haematoxylin and eosin (H&E) stained glass slides: a major step forward towards clinical translation

The Analyst ◽  
2017 ◽  
Vol 142 (8) ◽  
pp. 1258-1268 ◽  
Author(s):  
Michael J. Pilling ◽  
Alex Henderson ◽  
Jonathan H. Shanks ◽  
Michael D. Brown ◽  
Noel W. Clarke ◽  
...  
Keyword(s):  
Diagnostic Tool ◽  
Clinical Translation ◽  
Great Promise ◽  
Stained Glass ◽  
Major Step ◽  
Glass Slides ◽  
Important Diagnostic Tool ◽  
Infrared Spectral ◽  
Spectral Histopathology

Infrared spectral histopathology has shown great promise as an important diagnostic tool, with the potential to complement current pathological methods.

scholarly journals T-Scan III System Diagnostic Tool for Digital Occlusal Analysis in Orthodontics – a Modern Approach

PRILOZI ◽  
2014 ◽  
Vol 35 (2) ◽  
pp. 155-160 ◽  
Author(s):  
Vesna Trpevska ◽  
Gordana Kovacevska ◽  
Alberto Benedeti ◽  
Bozidar Jordanov
Keyword(s):  
Systematic Review ◽  
Diagnostic Tool ◽  
Reference Lists ◽  
Great Promise ◽  
Inclusion Criteria ◽  
Modern Approach ◽  
Diagnostic Screening ◽  
Randomized Controlled ◽  
Language Restriction ◽  
Screening Device

Abstract Introduction: This systematic literature review was performed to establish the mechanism, methodology, characteristics, clinical application and opportunities of the T-Scan III System as a diagnostic tool for digital occlusal analysis in different fields of dentistry, precisely in orthodontics. Methods: Searching of electronic databases, using MEDLINE and PubMed, hand searching of relevant key journals, and screening of reference lists of included studies with no language restriction was performed. Publications providing statistically examined data were included for systematic review. Results: Twenty potentially relevant Randomized Controlled Trials (RCTs) were identified. Only ten met the inclusion criteria. The literature demonstrates that using digital occlusal analysis with T-Scan III System in orthodontics has significant advantage with regard to the capability of measuring occlusal parameters in static positions and during dynamic of the mandible. Conclusion: Within the scope of this systematic review, there is evidence to support that T-Scan system is rapid and accurate in identifying the distribution of the tooth contacts and it shows great promise as a clinical diagnostic screening device for occlusion and for improving the occlusion after various dental treatments. Additional clinical studies are required to advance the indication filed of this system. Importance of using digital occlusal T-Scan analysis in orthodontics deserves further investigation.

scholarly journals Development of cyclic peptides with potent in vivo osteogenic activity through RaPID-based affinity maturation

2020 ◽  
Vol 117 (49) ◽  
pp. 31070-31077 ◽  
Author(s):  
Nasir K. Bashiruddin ◽  
Mikihito Hayashi ◽  
Masanobu Nagano ◽  
Yan Wu ◽  
Yukiko Matsunaga ◽  
...  
Keyword(s):  
Bone Resorption ◽  
Bone Formation ◽  
Cyclic Peptides ◽  
Treatment Options ◽  
Affinity Maturation ◽  
Great Promise ◽  
Anabolic Activity ◽  
Anabolic Treatment ◽  
Increase Bone Formation

Osteoporosis is caused by a disequilibrium between bone resorption and bone formation. Therapeutics for osteoporosis can be divided into antiresorptives that suppress bone resorption and anabolics which increase bone formation. Currently, the only anabolic treatment options are parathyroid hormone mimetics or an anti-sclerostin monoclonal antibody. With the current global increases in demographics at risk for osteoporosis, development of therapeutics that elicit anabolic activity through alternative mechanisms is imperative. Blockade of the PlexinB1 and Semaphorin4D interaction on osteoblasts has been shown to be a promising mechanism to increase bone formation. Here we report the discovery of cyclic peptides by a novel RaPID (Random nonstandard Peptides Integrated Discovery) system-based affinity maturation methodology that generated the peptide PB1m6A9 which binds with high affinity to both human and mouse PlexinB1. The chemically dimerized peptide, PB1d6A9, showed potent inhibition of PlexinB1 signaling in mouse primary osteoblast cultures, resulting in significant enhancement of bone formation even compared to non-Semaphorin4D–treated controls. This high anabolic activity was also observed in vivo when the lipidated PB1d6A9 (PB1d6A9-Pal) was intravenously administered once weekly to ovariectomized mice, leading to complete rescue of bone loss. The potent osteogenic properties of this peptide shows great promise as an addition to the current anabolic treatment options for bone diseases such as osteoporosis.

scholarly journals Identification and characterization of a cDNA and the gene encoding the mouse ubiquitously expressed glucose-6-phosphatase catalytic subunit-related protein

2004 ◽  
Vol 32 (1) ◽  
pp. 33-53 ◽  
Author(s):  
JN Boustead ◽  
CC Martin ◽  
JK Oeser ◽  
CA Svitek ◽  
SI Hunter ◽  
...  
Keyword(s):  
Fusion Gene ◽  
Gene Promoter ◽  
Catalytic Subunit ◽  
Mouse Tissue ◽  
Related Protein ◽  
Subunit Gene ◽  
Chromosome 11 ◽  
Gene Encoding ◽  
Identification And Characterization

Glucose-6-phosphatase (G6Pase) catalyzes the final step in the gluconeogenic and glycogenolytic pathways, the hydrolysis of glucose-6-phosphate (G6P) to glucose and phosphate. This paper describes the identification and characterization of a cDNA and the gene encoding the mouse ubiquitously expressed G6Pase catalytic subunit-related protein (UGRP). The open reading frame of this UGRP cDNA encodes a protein (346 amino acids (aa); Mr 38,755) that shares 36% overall identity (56% similarity) with the mouse G6Pase catalytic subunit (357 aa; Mr 40,454). UGRP exhibits a similar predicted transmembrane topology and conservation of many of the catalytically important residues with the G6Pase catalytic subunit; however, unlike the G6Pase catalytic subunit, UGRP does not catalyze G6P hydrolysis and does not contain a carboxy-terminal di-lysine endoplasmic reticulum retention signal. UGRP mRNA was detected by RNA blot analysis in every mouse tissue examined with the highest expression in heart, brain, testis and kidney. Database analysis showed that the mouse UGRP gene is composed of six exons, spans approximately 4.2 kbp of genomic DNA and is located on chromosome 11 along with the G6Pase catalytic subunit gene. The UGRP gene transcription start sites were mapped by primer extension analysis, and the activity of the mouse UGRP gene promoter was analyzed using luciferase fusion gene constructs. In contrast to the G6Pase catalytic subunit gene promoter, the UGRP promoter was highly active in all cell lines examined.

scholarly journals The HRX Proto-oncogene Product Is Widely Expressed in Human Tissues and Localizes to Nuclear Structures

Blood ◽  
1997 ◽  
Vol 89 (9) ◽  
pp. 3361-3370 ◽  
Author(s):  
Lisa H. Butler ◽  
Robert Slany ◽  
Xiangmin Cui ◽  
Michael L. Cleary ◽  
David Y. Mason
Keyword(s):  
Fusion Gene ◽  
Cell Types ◽  
Leukemic Cells ◽  
Chromosome Band ◽  
Lymphoid Tissues ◽  
Nuclear Staining ◽  
Human Tissues ◽  
Cell Nuclei ◽  
Reciprocal Translocations ◽  
Nuclear Structures

Abstract Chromosomal rearrangement of the HRX (MLL, ALL-1, Htrx) gene situated at chromosome band 11q23 is one of the most frequent genetic changes in infant leukemias of myeloid and lymphoid lineage and in treatment-induced secondary leukemias. The HRX gene codes for a predicted 431-kD protein that shows significant homology to the Drosophila trithorax protein, an Hox epigenetic regulator. Typically, the region encoding the HRX gene is rearranged, mostly in reciprocal translocations with a number of partners, resulting in a range of fusion genes. However, this is not the only abnormality affecting HRX because partial duplication of the gene, as well as interstitial deletions, can occur. Despite extensive studies of HRX at the genetic level, the protein products of the HRX gene and their patterns of expression in normal and leukemic cells remain uncharacterized. In this study we analyzed the distribution and localization of HRX proteins in cell lines and human tissues, using both polyclonal and monoclonal antibodies. The specificity of these reagents was confirmed using cells transfected with the HRX-ENL fusion gene. Western blot analyses of protein extracts from cells carrying the t(11; 19) and t(4; 11) translocations showed HRX chimeric proteins whose migrations corresponded to the sizes predicted from analyses of translocation-induced fusion mRNAs expressed by the derivative 11 chromosomes. Immunocytochemical analysis showed a punctate distribution of wild-type and chimeric HRX proteins within cell nuclei, suggesting that HRX localizes to nuclear structures in cells with and without 11q23 translocations. Nuclear staining was found in the majority of tissues studied with the strongest reactivity in cerebral cortex, kidney, thyroid, and lymphoid tissues. Thus, HRX is widely expressed in most cell types including hematopoietic cells, a finding that precludes an immunocytochemical approach for diagnosis of leukemias bearing 11q23 structural abnormalities.

scholarly journals Evaluation of NTRK Gene Fusion by Five Different Platforms in Triple-Negative Breast Carcinoma

2021 ◽  
Vol 8 ◽  
Author(s):  
Shafei Wu ◽  
Xiaohua Shi ◽  
Xinyu Ren ◽  
Kaimi Li ◽  
Junyi Pang ◽  
...  
Keyword(s):  
Breast Carcinoma ◽  
Tumor Cells ◽  
Targeted Therapies ◽  
Gene Fusion ◽  
Triple Negative ◽  
Fusion Gene ◽  
Rare Event ◽  
Nuclear Staining ◽  
Rt Pcr ◽  
Triple Negative Breast Carcinoma

Triple-negative breast carcinoma (TNBC) is an aggressive disease that has a poor prognosis since it lacks effective treatment methods. Neurotrophic tyrosine receptor kinase (NTRK) fusion genes are excellent candidates for targeted RTK inhibitor therapies and there are available targeted therapy drugs for the treatment of TRK fusion-positive tumors in a tumor agnostic pattern. Our study was designed to investigate the NTRK gene fusion status in TNBC patients and to determine whether RTK-targeted therapies are suitable for TNBC patients. A total of 305 TNBC patients were enrolled in our study. IHC was employed as a prescreening method, and IHC positive cases were further submitted for evaluation by FISH, RT-PCR, and NGS methods. NTRK IHC was evaluated successfully in 287 of the 305 cases, and there were 32 (11.15%) positive cases. FISH was carried out in the 32 IHC positive cases. There were 13 FISH-positive cases if the threshold was set as >15% of the 100 counted tumor cells having a split orange and green signal with more than one signal diameter. There were only 2 FISH-positive cases if the cutoff value was defined as >15% of the counted tumor cells having a split signal with more than two signal diameter widths. One of the FISH-positive cases had a separate NTRK3 FISH signal in 88% of the tumor cells, and its IHC result was strong nuclear staining in all the tumor cells. After evaluation of the morphology, it was re-diagnosed as secretory breast carcinoma, and the NGS result confirmed that it had a NTRK3-ETV6 fusion gene. The other FISH-positive cases were all negative for NTRK gene fusion in the NGS or RT-PCR examination. The NTRK gene fusion rate was low in our TNBC cohort. NTRK gene fusion may be a rare event in TNBC. The high false-positive rate of NTRK gene fusion detected by IHC questions its role as a prescreening method in TNBC. More data may be needed to determine a suitable threshold for NTRK FISH in TNBC in the future. More studies are needed to confirm whether RTK-targeted therapies are appropriate treatments for TNBC patients.

scholarly journals Fish Analysis for HEY1-NCOA2 Fusion Gene is Useful Diagnostic Tool for Mesenchymal Chondrosarcoma

2013 ◽  
Vol 24 ◽  
pp. ix72
Author(s):  
K. Ono ◽  
K. Takada ◽  
R. Takimoto ◽  
T. Sato ◽  
S. Iyama ◽  
...  
Keyword(s):  
Diagnostic Tool ◽  
Fusion Gene ◽  
Mesenchymal Chondrosarcoma ◽  
Fish Analysis

Automated cell counting of astrocytes on patterned substrates containing aliphatic and charged properties

1995 ◽  
Vol 53 ◽  
pp. 974-975
Author(s):  
W. Shain ◽  
H. Ancin ◽  
H.C. Craighead ◽  
M. Isaacson ◽  
L. Kam ◽  
...  
Keyword(s):  
Cell Culture ◽  
Cell Attachment ◽  
Culture Method ◽  
Cell Counting ◽  
Data Sets ◽  
Nuclear Staining ◽  
Double Positive ◽  
A Cell ◽  
Wafer Test ◽  
Cell Densities

Neural protheses have potential to restore nervous system functions lost by trauma or disease. Nanofabrication extends this approach to implants for stimulating and recording from single or small groups of neurons in the spinal cord and brain; however, tissue compatibility is a major limitation to their practical application. We are using a cell culture method for quantitatively measuring cell attachment to surfaces designed for nanofabricated neural prostheses.Silicon wafer test surfaces composed of 50-μm bars separated by aliphatic regions were fabricated using methods similar to a procedure described by Kleinfeld et al. Test surfaces contained either a single or double positive charge/residue. Cyanine dyes (diIC18(3)) stained the background and cell membranes (Fig 1); however, identification of individual cells at higher densities was difficult (Fig 2). Nuclear staining with acriflavine allowed discrimination of individual cells and permitted automated counting of nuclei using 3-D data sets from the confocal microscope (Fig 3). For cell attachment assays, LRM5 5 astroglial cells and astrocytes in primary cell culture were plated at increasing cell densities on test substrates, incubated for 24 hr, fixed, stained, mounted on coverslips, and imaged with a 10x objective.

Molecular and Microscopic Analysis of Altered Gene Expression in Transgenic and Gene Targeted Mice

1996 ◽  
Vol 54 ◽  
pp. 12-13
Author(s):  
W. K. Jones ◽  
J. Robbins
Keyword(s):  
Gene Expression ◽  
Pulmonary Veins ◽  
Microscopic Analysis ◽  
Mouse Tissue ◽  
Adult Heart ◽  
Heavy Chains ◽  
Altered Gene Expression ◽  
Altered Gene ◽  
Pulmonary Myocardium

Two myosin heavy chains (MyHC) are expressed in the mammalian heart and are differentially regulated during development. In the mouse, the α-MyHC is expressed constitutively in the atrium. At birth, the β-MyHC is downregulated and replaced by the α-MyHC, which is the sole cardiac MyHC isoform in the adult heart. We have employed transgenic and gene-targeting methodologies to study the regulation of cardiac MyHC gene expression and the functional and developmental consequences of altered α-MyHC expression in the mouse.We previously characterized an α-MyHC promoter capable of driving tissue-specific and developmentally correct expression of a CAT (chloramphenicol acetyltransferase) marker in the mouse. Tissue surveys detected a small amount of CAT activity in the lung (Fig. 1a). The results of in situ hybridization analyses indicated that the pattern of CAT transcript in the adult heart (Fig. 1b, top panel) is the same as that of α-MyHC (Fig. 1b, lower panel). The α-MyHC gene is expressed in a layer of cardiac muscle (pulmonary myocardium) associated with the pulmonary veins (Fig. 1c). These studies extend our understanding of α-MyHC expression and delimit a third cardiac compartment.

Are HOLZ lines kinematic in off-zone-axis orientation?

1993 ◽  
Vol 51 ◽  
pp. 692-693
Author(s):  
J. M. Zuo ◽  
A. L. Weickenmeier ◽  
R. Holmestad ◽  
J. C. H. Spence
Keyword(s):  
Structure Determination ◽  
Standard Procedure ◽  
High Order ◽  
Zone Axis ◽  
Great Promise ◽  
Line Position ◽  
Measured Intensity ◽  
Constant Intensity ◽  
Axis Orientation ◽  
Diffraction Condition

The application of high order reflections in a weak diffraction condition off the zone axis center, including those in high order laue zones (HOLZ), holds great promise for structure determination using convergent beam electron diffraction (CBED). It is believed that in this case the intensities of high order reflections are kinematic or two-beam like. Hence, the measured intensity can be related to the structure factor amplitude. Then the standard procedure of structure determination in crystallography may be used for solving unknown structures. The dynamic effect on HOLZ line position and intensity in a strongly diffracting zone axis is well known. In a weak diffraction condition, the HOLZ line position may be approximated by the kinematic position, however, it is not clear whether this is also true for HOLZ intensities. The HOLZ lines, as they appear in CBED patterns, do show strong intensity variations along the line especially near the crossing of two lines, rather than constant intensity along the Bragg condition as predicted by kinematic or two beam theory.

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