Presentation by Alvin Ayala Highlighting Luminex Technology
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The document outlines an experiment to determine the activation of a growth factor receptor (Factor X) in solid tumor cell lines using Luminex technology. Several tumor cell lines were screened for expression of the receptor via quantitative RT-PCR. HT-29 (colorectal) and FADU (head and neck) cell lines expressed the receptor and were chosen for further analysis. Stimulation of the cell lines with Factor X did not produce proliferation or demonstrate phosphorylation of the receptor via western blot. However, Luminex technology was able to detect downstream signaling through p-Akt in response to Factor X addition in the cell lines, showing it to be a more sensitive measure of receptor activation than other assays used.
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This study aimed to detect Mycoplasma gallisepticum (MG) strains in chicken flocks in Egypt with respiratory disease. Nasal swabs from 49 flocks were tested by semi-nested PCR and restriction fragment length polymorphism (RFLP) of the pvpA gene. MG was detected in 37 flocks (75.5% incidence). RFLP analysis identified genetic similarity between some field isolates and the F vaccine strain. Experimental infection of chickens found that field isolates and the F strain were transmitted between infected and contact birds based on PCR testing, with some contact birds developing antibodies. This highlights the need for improved diagnostics and control of MG variant strains.
Gene expression profile of the tumor microenvironment from 40 NSCLC FFPE and ...
Gene expression profile of the tumor microenvironment from 40 NSCLC FFPE and ...Thermo Fisher Scientific
The tumor microenvironment (TME) is the intersection between tumor cells and
surrounding non-transformed cells. It contains immune cells, signaling molecules,
stromal and extracellular matrix. Research has shown the TME is often associated
with tumor growth. However, the function and regulatory mechanism of each
constituent is still poorly understood. The presence of PD-L1 is a promising marker
to predict positive response for T cell checkpoint therapy. Current IHC methods to
measure PD-L1 are subjective and highly variable. A higher-throughput and
standardized method that can systematically measure gene expression of cells
present in the TME has emerged to be a more desirable solution.
We applied the OncomineTM Immune Response Research Assay to measure the
expression of 395 genes in non-small cell lung cancer (NSCLC) research samples
from 40 matched FFPE and fresh frozen sample types. This assay covers genes
involved in checkpoint pathway, T cell regulation, cytokine and interferon signaling
pathways, and markers of different tumor infiltrating lymphocyte (TIL) subsets, as
well as tumor markers. With an input requirement of 10 ng of total RNA, libraries
were generated, templated on the Ion ChefTM and sequenced on the Ion S5TM
System. Sequencing data was analyzed and mapped with Torrent Suite Software
and differential expression analysis was conducted with AffymetrixTM Transcriptome
Analysis Console.PD1_PDL1_Short Paper
This document summarizes two scientific articles about Programmed Death-1 (PD-1) and its ligands PD-L1 and PD-L2. The first article analyzes the basal and induced expression of PD-1 ligands in three chordoma cell lines and finds that while basal expression is low, expression is upregulated by IFN-γ but not IL-4. The second article examines blocking the PD-1/PD-L1 pathway to prevent loss of effector function in adoptively transferred central memory CD8+ T cells against tumors.
Src jbbr-20-120 Dr. ihsan edan abdulkareem alsaimary PROFESSOR IN MEDICAL M...
Dr. ihsan edan abdulkareem alsaimary
PROFESSOR IN MEDICAL MICROBIOLOGY AND MOLECULAR IMMUNOLOGY
ihsanalsaimary@gmail.com
mobile : 009647801410838
university of basrah - college of medicine - basrah -IRAQ
FGFBP1 pathways control after induction of a conditional transgene in a mouse...
A systems biology approach to analyzing large data sets, such as this study which involved five full mouse cDNA arrays allows the researcher to capture a snapshot of the unfolding remodeling events of an organisms response to change, stress or disease. Analyzing data in this form involves filtering the biological signal from the noise. Sorting the noise in appropriate manners is essential to be able to complete the biological story. Building on existing knowledge base, we can complete the picture as long as the proper context of the collection, normalization and analysis is maintained. High throughput technologies such as microarrays and RNA sequencing as enabled by next generation sequencing presents the researcher with the challenge of extracting meaningful information from the measurements. Software tools and analysis techniques are not a substitute to understanding the biological context from which the data are collected. Engineering and digital signal processing has allowed us to derive the understanding of how to reconstruct a signal from the presence of a continual stream of noisy analog data. Sampling frequency and proper filtering are a must to be able to sort out a meaningful signal from the noise. These same principles apply not only to communication theory but also when studying large data such as those that may be collected from high throughput systems such as a Affymetrix mouse cDNA array.
Mol Pharmacol-2003-Jones-471-7
This document describes the identification and characterization of a novel G protein-coupled receptor (GPCR) termed R527 that is highly expressed on human eosinophils and neutrophils. R527 was found to bind the eicosanoid 5-oxo-6E,8Z,11Z,14Z-eicosatetraenoic acid (5-oxo-ETE) and mediate its inflammatory effects. Stable cell lines expressing R527 were used to screen ligands and identified 5-oxo-ETE as an agonist. R527 showed pharmacological properties similar to the previously described 5-oxo-ETE receptor on immune cells. The identification of R527 provides insight into the physiological role
Cell-based Reporter Assays: Measure 45 Signaling Pathway Activity in Any Cel...
Would you like to measure signaling pathway activity in your favorite cell? Learn how to successfully apply convenient and robust reporter assays to your RNA interference, gene over-expression, protein, or small molecule studies. The Cignal Reporter Assays are an excellent tool for studying pathway signaling activity in cells that are amenable to transfection, available for studying numerous pathways including (ROS, Wnt, NF-kB, Notch, cAMP/PKA, TGFbeta, and the Cignal Lenti Reporter Assays combines the power of a lentiviral delivery system with our robust transcription factor reporter technology, enabling you to study signal pathways in virtually any cell type. You can find a technology overview, protocol tutorial, and application examples in the following presentation.
Cell-based Reporter Assays: Measure 45 Signaling Pathway Activity in Any Cell...
Would you like to measure signaling pathway activity in your favorite cell? Learn how to successfully apply convenient and robust reporter assays to your RNA interference, gene over-expression, protein, or small molecule studies. The Cignal Reporter Assays are an excellent tool for studying pathway signaling activity in cells that are amenable to transfection, available for studying numerous pathways including (ROS, Wnt, NF-kB, Notch, cAMP/PKA, TGFbeta, and the Cignal Lenti Reporter Assays combines the power of a lentiviral delivery system with our robust transcription factor reporter technology, enabling you to study signal pathways in virtually any cell type. You can find a technology overview, protocol tutorial, and application examples in the following presentation.
AJP_12-0313_Araten_et_al_Word_Version
This document describes a study that analyzed the frequency of cells with a PIG-A mutant phenotype (PNH phenotype) in samples from 19 patients with acute lymphoblastic leukemia (ALL) and immortalized B cell lines (BLCLs) from healthy donors. The BLCLs showed a median frequency of 11 x 10-6, while the ALL samples showed two distinct populations - about half of samples had a median frequency of 13 x 10-6, similar to controls, while the other half had a significantly higher median frequency of 566 x 10-6. This suggests that in ALL, there are two phenotypes with respect to hypermutability, which may correlate with the number of mutations required to cause the leukemia.
COMPARISON OF ANTIGEN DETECTION METHODS OF PESTE DES PETITS RUMINANTS VIRUS I...
This study compared three methods for detecting the antigen of peste des petits ruminants virus (PPRV) in clinical samples from small ruminants: agar gel immunodiffusion (AGID), haemagglutination (HA) test, and immuno-capture enzyme-linked immunosorbent assay (IC-ELISA). AGID detected antigen in 8.95% of samples, HA detected it in 20.9% of samples, and IC-ELISA detected it in 34.3% of samples, showing IC-ELISA to be the most sensitive. There was no significant agreement between the tests according to kappa statistics. The rapid detection of viral antigen through appropriate methods can help diagnose infection earlier and
sclabas2003
This document discusses a study that investigated how inhibiting the epidermal growth factor receptor (EGFR) signaling pathway with the anti-EGFR monoclonal antibody IMC-C225 affects nuclear factor-kappa B (NF-κB) activation and regulation of apoptosis genes in human pancreatic cancer cells. The study found that IMC-C225 treatment blocked EGFR activation in pancreatic cancer cells, leading to decreased NF-κB DNA binding activity. This downregulation of NF-κB by IMC-C225 resulted in decreased expression of the anti-apoptotic genes bcl-xl and bfl-1. Therefore, targeting the NF-κB pathway with an anti-EGFR antibody may help restore apoptosis in pancreatic cancer cells and
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Doi10.18535ijmsciv7i11.06 Dr. ihsan edan abdulkareem alsaimary PROFESSOR I...
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Gene expression profile of the tumor microenvironment from 40 NSCLC FFPE and ...
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Src jbbr-20-120 Dr. ihsan edan abdulkareem alsaimary PROFESSOR IN MEDICAL M...
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Folding is a recent technique for building efficient recursive SNARKs. Several elegant folding protocols have been proposed, such as Nova, Supernova, Hypernova, Protostar, and others. However, all of them rely on an additively homomorphic commitment scheme based on discrete log, and are therefore not post-quantum secure. In this work we present LatticeFold, the first lattice-based folding protocol based on the Module SIS problem. This folding protocol naturally leads to an efficient recursive lattice-based SNARK and an efficient PCD scheme. LatticeFold supports folding low-degree relations, such as R1CS, as well as high-degree relations, such as CCS. The key challenge is to construct a secure folding protocol that works with the Ajtai commitment scheme. The difficulty, is ensuring that extracted witnesses are low norm through many rounds of folding. We present a novel technique using the sumcheck protocol to ensure that extracted witnesses are always low norm no matter how many rounds of folding are used. Our evaluation of the final proof system suggests that it is as performant as Hypernova, while providing post-quantum security.
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Presentation by Alvin Ayala Highlighting Luminex Technology
- 1. Utility of Luminex technology to determine growth factor activation and downstream signaling of factor (X) Alvin Ayala Amgen Inc April 2006
- 3. Experimental approach Screen for human tumor lines that express the receptor for factor X Receptor gene expression profiling Design primers and probes For mRNA detection In vitro proliferation assays adding factor X In vitro IP/Western blot of p-Y of the receptor after stimulation Confirm downstream signaling triggered by factor addition
- 5. Human tumor cell line gene expression of receptor X
- 7. HT-29 cells response to factor X
- 8. FADU cells response to factor X In vitro proliferation poor
- 10. Assessing downstream signaling P-Akt status after factor addition Experimental design 5 10 20 30 50 60 GF addition 100ng/ml Cells plated 5 million/well Serum starvation 48 hours Assay Cell lysis Time (mins)
- 11. Assessing downstream signaling p-Akt status after factor X addition
- 14. Thank You (science rules!)