A summary presentation of my scientific work.
My laboratory focused on an enzyme KDM5b (aka PLU-1, JARID1b) that was widely expressed during development and played a key role in progression of breast cancer through HER-2.
My lab focused on understanding the key biochemical activity of the enzyme through dissecting the proteomic and genomic interactors.
Our results were confirmed through the use of ES cells, adult stem cells and mouse models.
Much of this work remains unpublished, please contact me for more information and/or access to any reagents that I still have as part of this work.
crwynder@gmail.com
Regulation of KDM5 by multiple cofactors regulates cancer and stem cellsChristopher Wynder
Presentation of data regarding proteins that regulate the activity of KDM5b.
The studies use multiple disciplines including in vitro enzymology, ES cell studies of differentiation, Mass spectrometry to detect protein protein interactions.
These studies resulted in a comprehensive view of KDM5b function. It required development of at least three novel assays that are focused on moving epigenetic research from yeast and HeLa cell types to primary, clinically relevant cell types.
The techniques have been successfully used in Embryonic stem cells (human and mouse), Neural stem cells (mouse and patient derived as well as iPSCs.
Regulation of KDM5 by multiple cofactors regulates cancer and stem cellsChristopher Wynder
Presentation of data regarding proteins that regulate the activity of KDM5b.
The studies use multiple disciplines including in vitro enzymology, ES cell studies of differentiation, Mass spectrometry to detect protein protein interactions.
These studies resulted in a comprehensive view of KDM5b function. It required development of at least three novel assays that are focused on moving epigenetic research from yeast and HeLa cell types to primary, clinically relevant cell types.
The techniques have been successfully used in Embryonic stem cells (human and mouse), Neural stem cells (mouse and patient derived as well as iPSCs.
Epigenetics and it's relevance in crop improvementShamlyGupta
Epigenetics means ‘above’ or ‘on top of genetics’
A study of the changes in gene expression that are mitotically and/or meiotically heritable and do not involve a change in the DNA sequence
Gene-regulatory information that is not expressed in DNA sequences but transmitted from one generation (of cells or organisms) to the next
Coined by embryologist C. H. Waddington in 1942.
Plant epigenetic memory in plant growth behavior and stress response. Sally M...CIAT
Speaker: Sally Mackenzie, Lloyd and Dottie Huck Chair for Functional Genomics, Department of Biology, Pennsylvania State University. Fellow in the American Society of Plant Biologists and the American Association for the Advancement of Science (AAAS).
Event: Robert D. Havener Seminar on “Innovations for Crop Productivity”.
http://ciat.cgiar.org/event/robert-d-havener-seminar-on-innovations-for-crop-productivity/
Epigenetics can be used to explain the phenomena that cannot be explained by genetics/genomics, such as the differences between monozygotic twins, which are considered to be genetically identical.
The Role of DNA Methylation in Coronary Artery DiseaseBardia Farivar
Epigenetic studies have identified DNA methylation in coronary artery disease (CAD). How the critical genes interact at the cellular level to cause CAD is still unknown. The discovery of DNA methylation inspired researchers to explore relationships in genomic coding and disease phenotype. In the past two decades, there have been many findings regarding the relationship between DNA methylation and CAD development, and the DNA methylation of critical genes have been found to be significantly changed during CAD, including DNA methylation at homocysteine, Alu and long Interspersed Element 1 (LINE-1) repetitive elements.
Ibica2014 p(8) visualizing and identifying the dna methylationAboul Ella Hassanien
DNA methylation is an epigenetic mechanism that cells use to control
gene expression. DNA methylation has become one of the hottest topics in cancer
research, especially for abnormally hypermethylated tumor suppressor genes
or hypomethylaed oncogenes research. The analysis of DNA methylation data
determines the differential hypermethlated or hypomethylated genes that are candidate
to be cancer biomarkers. Visualization the DNA methylation status may
lead to discover new relationships between hypomethylated and hypermethylated
genes, therefore this paper applied a mathematical modelling theory called formal
concept analysis for visualizing DNA methylation status.
The epigenetic regulation of DNA-templated processes has been intensely studied over the last 15
years. DNA methylation, histone modification, nucleosome remodeling, and RNA-mediated targeting regulate many biological processes that are fundamental to the genesis of cancer. Here, we
present the basic principles behind these epigenetic pathways and highlight the evidence suggesting that their misregulation can culminate in cancer. This information, along with the promising clinical and preclinical results seen with epigenetic drugs against chromatin regulators, signifies that it
is time to embrace the central role of epigenetics in cancer.
Epigenetics and it's relevance in crop improvementShamlyGupta
Epigenetics means ‘above’ or ‘on top of genetics’
A study of the changes in gene expression that are mitotically and/or meiotically heritable and do not involve a change in the DNA sequence
Gene-regulatory information that is not expressed in DNA sequences but transmitted from one generation (of cells or organisms) to the next
Coined by embryologist C. H. Waddington in 1942.
Plant epigenetic memory in plant growth behavior and stress response. Sally M...CIAT
Speaker: Sally Mackenzie, Lloyd and Dottie Huck Chair for Functional Genomics, Department of Biology, Pennsylvania State University. Fellow in the American Society of Plant Biologists and the American Association for the Advancement of Science (AAAS).
Event: Robert D. Havener Seminar on “Innovations for Crop Productivity”.
http://ciat.cgiar.org/event/robert-d-havener-seminar-on-innovations-for-crop-productivity/
Epigenetics can be used to explain the phenomena that cannot be explained by genetics/genomics, such as the differences between monozygotic twins, which are considered to be genetically identical.
The Role of DNA Methylation in Coronary Artery DiseaseBardia Farivar
Epigenetic studies have identified DNA methylation in coronary artery disease (CAD). How the critical genes interact at the cellular level to cause CAD is still unknown. The discovery of DNA methylation inspired researchers to explore relationships in genomic coding and disease phenotype. In the past two decades, there have been many findings regarding the relationship between DNA methylation and CAD development, and the DNA methylation of critical genes have been found to be significantly changed during CAD, including DNA methylation at homocysteine, Alu and long Interspersed Element 1 (LINE-1) repetitive elements.
Ibica2014 p(8) visualizing and identifying the dna methylationAboul Ella Hassanien
DNA methylation is an epigenetic mechanism that cells use to control
gene expression. DNA methylation has become one of the hottest topics in cancer
research, especially for abnormally hypermethylated tumor suppressor genes
or hypomethylaed oncogenes research. The analysis of DNA methylation data
determines the differential hypermethlated or hypomethylated genes that are candidate
to be cancer biomarkers. Visualization the DNA methylation status may
lead to discover new relationships between hypomethylated and hypermethylated
genes, therefore this paper applied a mathematical modelling theory called formal
concept analysis for visualizing DNA methylation status.
The epigenetic regulation of DNA-templated processes has been intensely studied over the last 15
years. DNA methylation, histone modification, nucleosome remodeling, and RNA-mediated targeting regulate many biological processes that are fundamental to the genesis of cancer. Here, we
present the basic principles behind these epigenetic pathways and highlight the evidence suggesting that their misregulation can culminate in cancer. This information, along with the promising clinical and preclinical results seen with epigenetic drugs against chromatin regulators, signifies that it
is time to embrace the central role of epigenetics in cancer.
Histone H2B is one of the 5 main histone proteins involved in the structure of chromatin in eukaryotic cells.
Histone H2B is a structural protein that helps organise eukaryotic DNA. It plays an important role in the biology of the nucleus where it is involved in the packaging and maintaining of chromosomes, regulation of transcription, and replication and repair of DNA. Histone H2B helps regulate chromatin structure and function through post-translational modifications and specialised histone variants.
The antibody detects endogenous Histone H2B (Tri Methyl Lys43) protein.
Anti-Histone H2B (Tri Methyl Lys43) antibody - http://www.stjohnslabs.com/histone-h2b-tri-methyl-lys43-antibody?filter_name=STJ97165
Join our Antibody Validation Project - http://www.stjohnslabs.com/services/antibody-validation
Histone H2B is one of the 5 main histone proteins involved in the structure of chromatin in eukaryotic cells.
Histone H2B is a structural protein that helps organise eukaryotic DNA. It plays an important role in the biology of the nucleus where it is involved in the packaging and maintaining of chromosomes, regulation of transcription, and replication and repair of DNA. Histone H2B helps regulate chromatin structure and function through post-translational modifications and specialised histone variants.
The antibody detects endogenous Histone H2B (Di Methyl Lys43) protein.
Anti-Histone H2B (Di Methyl Lys43) Antibody- http://www.stjohnslabs.com/histone-h2b-di-methyl-lys43-antibody?filter_name=STJ97164
Join our Antibody Validation Project - http://www.stjohnslabs.com/services/antibody-validation
Presentation entitled "Hit identification Strategies for Epigenetic Targets" at X-Gen Epigenetics iV, March 5-7th, 2012. Presentation was delivered by Dr Amy Quinn as I had a conflict which prevented my attendance
Conferencia de la Dra. Ana María Roa, Bióloga Molecular, sobre Epigenética, impartida en la Universidad Popular Carmen de Michelena de Tres Cantos el 1 de marzo de 2013.
Más información en:
http://www.universidadpopularc3c.es/index.php/actividades/conferencias/event/448-conferencia-una-revision-de-los-conocimientos-fundamentales-de-la-biologia-de-la-celula-la-epigenetica
DNA and Cell Reprogramming via Epigenetic Information Delivered by Magnetic Fields, Sound Vibrations and Coherent Water.
Don't Miss Dr. Carlo Ventura's presentation at the 2016 Conference for Consciousness & Human Evolution, a series of talks designed to give you tools to develop your consciousness. Click Here to Learn More
genes addiion\deeion\ediionthat lead to a therapeutic, prophylactic or diagnostic effect
Plasmid DNA
•Viral vectors
•Genetically engineered micro-organisms
•Human gene-editing technology
•Patient-derived cellular gene therapy products
The number of sequenced genes having unknown function continues to climb with the continuing decrease in the cost of genome sequencing. In Reverse Genetics (RG), functions of known genes are investigated with targeted modulation of gene activity, and hypothesis regarding gene function directly tested in vivo. Several RG approaches like insertional mutagenesis, fast neutron mutagenesis, TILLING and RNA interference have led to the identification of mutations in candidate genes and subsequent phenotypic analysis of these mutants.
Okabe et al. (2011) employed TILLING technique to screen six ethylene receptor genes in tomato (SlETR1–SlETR6) and two allelic mutants of SlETR1 (Sletr1-1 and Sletr1-2) with reduced ethylene response were identified. Using fast neutron mutagenesis, Li et al. (2001) obtained arabidopsis deletion mutants for bZIP transcription factor viz. AHBP 1b and OBF 5, a key regulator for systemic acquired resistance but their role were compensated by other regulatory factors in mutants. Terada et al. (2007) successfully blocked the expression of the Adh 2 gene through homologous recombination followed by transgenesis in rice however phenotype could not be determined since no differences were observed between wild and transgenic plants. RNA interference (RNAi) works as sequence-specific gene regulation and has been used in determination of function of many genes. Saurabh et al. (2014) reviewed the impact of RNAi in crop improvement and found its application in improvement of nutritional aspects, biotic and abiotic stresses, morphol¬ogy, crafting male sterility, enhanced secondary metabolite synthesis.
In addition, new advances in technology and reduction in sequencing cost may soon make it practical to use whole genome sequencing or gene targeting like ZFN technology and TAL effectors technology on a routine basis to identify or generate mutations in specific genes. Scholze and Boch (2011) mentioned that TAL effectors technology is more specific and predictable than ZFN. RG techniques have their own advantages and disadvantages depending on the species being targeted and the questions being addressed. Finally, with the continuous development of new technologies, the most efficient RG technique in the future may involve high throughput direct sequencing of part or complete genomes of individual plants followed by efficient novel tools to determine the function for utilization in crop improvement.
Oncology: Spatial Localization of Ras proteinsNachiket Vartak
This is a presentation of work done at the MPI Dortmund from 2008-2013 on the mechanism through with localization of the Ras protein in generated in cells. It presents the inhibiton Palmostatin-B, which inhibits this mechanism, leading to reveral of oncogenic signaling and cancerous phenotypes.
Similar to KDM5 epigenetic modifiers as a focus for drug discovery (20)
Whitepaper developed with Pharma Exec magazine on how EIM- Enterprise Information Management- can provide efficiency and kick start innovation by ensuring information flows correctly inside- and outside- the company
Healthcare products suffer from a lack of ability to control documents and non-clinical images. OpenText ApplicationXtender can solve that problem for vendors through our OEM program. This whitepaper goes through the benefits of embedding ApplicationXtender into healthcare products.
OpenText ApplicationXtender provides cost effective document management. For software vendors looking to expand or build a healthcare focused product, "AX" can be embedded to provide first class content services in without the high cost of research and development.
Automating Patient Management with ApplicationXtender WorkflowChristopher Wynder
The hardest part about managing a clinic is keeping everybody up-to-date with the right information. Whether this is simply making sure billing is alerted of a new bill or as complex as managing follow-ups after a referral. There is simply too many documents, emails and schedules for a person to manage. This is the value of workflow to a clinic or hospital- setting the rules regarding who gets to see certain types of documents and ensuring that know about the updated information.
Healthcare information management is complex. Not only does it require a system designed specifically for medically relevant information, you also need a system that manages billing and the normal every-business kind of HR, vendor, etc records that confound organizations of all kinds.
In this whitepaper we go through how Content Services (nee ECM) can be used as the connective tissue between your clinical systems and administrative use cases.
This deck goes through the Information conundrum and how ApplicationXtender is positioned to provide the technical platform for organizations to start moving from paper to a digital future
ThinkDox talk from ECNO 2017 on using Laserfiche to manage student records and student information. We use the examples of Field trip forms and student record search to highlight the potential administrative efficiencies that can be gained.
Information Management aaS AIIM First Canadian presentationChristopher Wynder
High level talk given at AIIM Canada's breakfast event March 23, 2017.
The talk goes through the challenges of information management in the era of BYOD and cloud services. The last part of the talk is how to start with a small but impactful project to show the value of IMaaS.
Histone demethylase and it srole in cell biology reviewChristopher Wynder
This document provides a scientific review of the histone demethylase enzymes; particularly the H3K4 demethlases (KDM5 family) focusing on their role in cell biology. This review was written in 2014
ECNO 2016-Using ECM to gain administrative efficiency for school boardsChristopher Wynder
Presentation from ECNO 2016. The presentation centers on embedding records management into process management. We take a IT project centric view of how to move from chaos to manage-able information access points. A key concept is how ECM and EIM technologies provide opportunities for school boards to reduce their costs and risk.
Embedding records management practices into how people work is essential to ensuring compliance and reducing risk in the digital age. This presentation goes through examples of how current processes and the mixed digital paper processes commonly found at organizations are reducing control rather than increasing it.
We are often why use a VAR- what am I paying you for? This presentation goes through the basics of how we implement Laserfiche and provide continual support above and beyond basic technical support, we make sure you understand what is possible and support you as you maximize your investment.
Laserfiche10 highlights- how the new features can benefit your mobile and wor...Christopher Wynder
Laserfiche 10 brings a lot of additional features for information management, workflow building and mobile content access. This slide deck provides the overview of how Laserfiche 10 can benefit clients looking to automate their processes.
Integrating user needs into ECM projects is key to success. Whether it is a initial implementation or a reboot or just expanding use, user needs and UX testing should be integrated into every project
Moving records management from a paper based strategy to a electronic strategy requires re-thinking what needs to be protected and where the threats to security exist.
The key is to stop focusing on the artifact (the document) and focus on the information that is important. Documents are just the storage media to move the information from person to person.
Prix Galien International 2024 Forum ProgramLevi Shapiro
June 20, 2024, Prix Galien International and Jerusalem Ethics Forum in ROME. Detailed agenda including panels:
- ADVANCES IN CARDIOLOGY: A NEW PARADIGM IS COMING
- WOMEN’S HEALTH: FERTILITY PRESERVATION
- WHAT’S NEW IN THE TREATMENT OF INFECTIOUS,
ONCOLOGICAL AND INFLAMMATORY SKIN DISEASES?
- ARTIFICIAL INTELLIGENCE AND ETHICS
- GENE THERAPY
- BEYOND BORDERS: GLOBAL INITIATIVES FOR DEMOCRATIZING LIFE SCIENCE TECHNOLOGIES AND PROMOTING ACCESS TO HEALTHCARE
- ETHICAL CHALLENGES IN LIFE SCIENCES
- Prix Galien International Awards Ceremony
Tom Selleck Health: A Comprehensive Look at the Iconic Actor’s Wellness Journeygreendigital
Tom Selleck, an enduring figure in Hollywood. has captivated audiences for decades with his rugged charm, iconic moustache. and memorable roles in television and film. From his breakout role as Thomas Magnum in Magnum P.I. to his current portrayal of Frank Reagan in Blue Bloods. Selleck's career has spanned over 50 years. But beyond his professional achievements. fans have often been curious about Tom Selleck Health. especially as he has aged in the public eye.
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Introduction
Many have been interested in Tom Selleck health. not only because of his enduring presence on screen but also because of the challenges. and lifestyle choices he has faced and made over the years. This article delves into the various aspects of Tom Selleck health. exploring his fitness regimen, diet, mental health. and the challenges he has encountered as he ages. We'll look at how he maintains his well-being. the health issues he has faced, and his approach to ageing .
Early Life and Career
Childhood and Athletic Beginnings
Tom Selleck was born on January 29, 1945, in Detroit, Michigan, and grew up in Sherman Oaks, California. From an early age, he was involved in sports, particularly basketball. which played a significant role in his physical development. His athletic pursuits continued into college. where he attended the University of Southern California (USC) on a basketball scholarship. This early involvement in sports laid a strong foundation for his physical health and disciplined lifestyle.
Transition to Acting
Selleck's transition from an athlete to an actor came with its physical demands. His first significant role in "Magnum P.I." required him to perform various stunts and maintain a fit appearance. This role, which he played from 1980 to 1988. necessitated a rigorous fitness routine to meet the show's demands. setting the stage for his long-term commitment to health and wellness.
Fitness Regimen
Workout Routine
Tom Selleck health and fitness regimen has evolved. adapting to his changing roles and age. During his "Magnum, P.I." days. Selleck's workouts were intense and focused on building and maintaining muscle mass. His routine included weightlifting, cardiovascular exercises. and specific training for the stunts he performed on the show.
Selleck adjusted his fitness routine as he aged to suit his body's needs. Today, his workouts focus on maintaining flexibility, strength, and cardiovascular health. He incorporates low-impact exercises such as swimming, walking, and light weightlifting. This balanced approach helps him stay fit without putting undue strain on his joints and muscles.
Importance of Flexibility and Mobility
In recent years, Selleck has emphasized the importance of flexibility and mobility in his fitness regimen. Understanding the natural decline in muscle mass and joint flexibility with age. he includes stretching and yoga in his routine. These practices help prevent injuries, improve posture, and maintain mobilit
These lecture slides, by Dr Sidra Arshad, offer a quick overview of physiological basis of a normal electrocardiogram.
Learning objectives:
1. Define an electrocardiogram (ECG) and electrocardiography
2. Describe how dipoles generated by the heart produce the waveforms of the ECG
3. Describe the components of a normal electrocardiogram of a typical bipolar leads (limb II)
4. Differentiate between intervals and segments
5. Enlist some common indications for obtaining an ECG
Study Resources:
1. Chapter 11, Guyton and Hall Textbook of Medical Physiology, 14th edition
2. Chapter 9, Human Physiology - From Cells to Systems, Lauralee Sherwood, 9th edition
3. Chapter 29, Ganong’s Review of Medical Physiology, 26th edition
4. Electrocardiogram, StatPearls - https://www.ncbi.nlm.nih.gov/books/NBK549803/
5. ECG in Medical Practice by ABM Abdullah, 4th edition
6. ECG Basics, http://www.nataliescasebook.com/tag/e-c-g-basics
Ozempic: Preoperative Management of Patients on GLP-1 Receptor Agonists Saeid Safari
Preoperative Management of Patients on GLP-1 Receptor Agonists like Ozempic and Semiglutide
ASA GUIDELINE
NYSORA Guideline
2 Case Reports of Gastric Ultrasound
Factory Supply Best Quality Pmk Oil CAS 28578–16–7 PMK Powder in Stockrebeccabio
Factory Supply Best Quality Pmk Oil CAS 28578–16–7 PMK Powder in Stock
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Communicate promptly and reply
Free of customs clearance, Double Clearance 100% pass delivery to USA, Canada, Spain, Germany, Netherland, Poland, Italy, Sweden, UK, Czech Republic, Australia, Mexico, Russia, Ukraine, Kazakhstan.Door to door service
Hot Selling Organic intermediates
Title: Sense of Smell
Presenter: Dr. Faiza, Assistant Professor of Physiology
Qualifications:
MBBS (Best Graduate, AIMC Lahore)
FCPS Physiology
ICMT, CHPE, DHPE (STMU)
MPH (GC University, Faisalabad)
MBA (Virtual University of Pakistan)
Learning Objectives:
Describe the primary categories of smells and the concept of odor blindness.
Explain the structure and location of the olfactory membrane and mucosa, including the types and roles of cells involved in olfaction.
Describe the pathway and mechanisms of olfactory signal transmission from the olfactory receptors to the brain.
Illustrate the biochemical cascade triggered by odorant binding to olfactory receptors, including the role of G-proteins and second messengers in generating an action potential.
Identify different types of olfactory disorders such as anosmia, hyposmia, hyperosmia, and dysosmia, including their potential causes.
Key Topics:
Olfactory Genes:
3% of the human genome accounts for olfactory genes.
400 genes for odorant receptors.
Olfactory Membrane:
Located in the superior part of the nasal cavity.
Medially: Folds downward along the superior septum.
Laterally: Folds over the superior turbinate and upper surface of the middle turbinate.
Total surface area: 5-10 square centimeters.
Olfactory Mucosa:
Olfactory Cells: Bipolar nerve cells derived from the CNS (100 million), with 4-25 olfactory cilia per cell.
Sustentacular Cells: Produce mucus and maintain ionic and molecular environment.
Basal Cells: Replace worn-out olfactory cells with an average lifespan of 1-2 months.
Bowman’s Gland: Secretes mucus.
Stimulation of Olfactory Cells:
Odorant dissolves in mucus and attaches to receptors on olfactory cilia.
Involves a cascade effect through G-proteins and second messengers, leading to depolarization and action potential generation in the olfactory nerve.
Quality of a Good Odorant:
Small (3-20 Carbon atoms), volatile, water-soluble, and lipid-soluble.
Facilitated by odorant-binding proteins in mucus.
Membrane Potential and Action Potential:
Resting membrane potential: -55mV.
Action potential frequency in the olfactory nerve increases with odorant strength.
Adaptation Towards the Sense of Smell:
Rapid adaptation within the first second, with further slow adaptation.
Psychological adaptation greater than receptor adaptation, involving feedback inhibition from the central nervous system.
Primary Sensations of Smell:
Camphoraceous, Musky, Floral, Pepperminty, Ethereal, Pungent, Putrid.
Odor Detection Threshold:
Examples: Hydrogen sulfide (0.0005 ppm), Methyl-mercaptan (0.002 ppm).
Some toxic substances are odorless at lethal concentrations.
Characteristics of Smell:
Odor blindness for single substances due to lack of appropriate receptor protein.
Behavioral and emotional influences of smell.
Transmission of Olfactory Signals:
From olfactory cells to glomeruli in the olfactory bulb, involving lateral inhibition.
Primitive, less old, and new olfactory systems with different path
MANAGEMENT OF ATRIOVENTRICULAR CONDUCTION BLOCK.pdfJim Jacob Roy
Cardiac conduction defects can occur due to various causes.
Atrioventricular conduction blocks ( AV blocks ) are classified into 3 types.
This document describes the acute management of AV block.
The prostate is an exocrine gland of the male mammalian reproductive system
It is a walnut-sized gland that forms part of the male reproductive system and is located in front of the rectum and just below the urinary bladder
Function is to store and secrete a clear, slightly alkaline fluid that constitutes 10-30% of the volume of the seminal fluid that along with the spermatozoa, constitutes semen
A healthy human prostate measures (4cm-vertical, by 3cm-horizontal, 2cm ant-post ).
It surrounds the urethra just below the urinary bladder. It has anterior, median, posterior and two lateral lobes
It’s work is regulated by androgens which are responsible for male sex characteristics
Generalised disease of the prostate due to hormonal derangement which leads to non malignant enlargement of the gland (increase in the number of epithelial cells and stromal tissue)to cause compression of the urethra leading to symptoms (LUTS
Title: Sense of Taste
Presenter: Dr. Faiza, Assistant Professor of Physiology
Qualifications:
MBBS (Best Graduate, AIMC Lahore)
FCPS Physiology
ICMT, CHPE, DHPE (STMU)
MPH (GC University, Faisalabad)
MBA (Virtual University of Pakistan)
Learning Objectives:
Describe the structure and function of taste buds.
Describe the relationship between the taste threshold and taste index of common substances.
Explain the chemical basis and signal transduction of taste perception for each type of primary taste sensation.
Recognize different abnormalities of taste perception and their causes.
Key Topics:
Significance of Taste Sensation:
Differentiation between pleasant and harmful food
Influence on behavior
Selection of food based on metabolic needs
Receptors of Taste:
Taste buds on the tongue
Influence of sense of smell, texture of food, and pain stimulation (e.g., by pepper)
Primary and Secondary Taste Sensations:
Primary taste sensations: Sweet, Sour, Salty, Bitter, Umami
Chemical basis and signal transduction mechanisms for each taste
Taste Threshold and Index:
Taste threshold values for Sweet (sucrose), Salty (NaCl), Sour (HCl), and Bitter (Quinine)
Taste index relationship: Inversely proportional to taste threshold
Taste Blindness:
Inability to taste certain substances, particularly thiourea compounds
Example: Phenylthiocarbamide
Structure and Function of Taste Buds:
Composition: Epithelial cells, Sustentacular/Supporting cells, Taste cells, Basal cells
Features: Taste pores, Taste hairs/microvilli, and Taste nerve fibers
Location of Taste Buds:
Found in papillae of the tongue (Fungiform, Circumvallate, Foliate)
Also present on the palate, tonsillar pillars, epiglottis, and proximal esophagus
Mechanism of Taste Stimulation:
Interaction of taste substances with receptors on microvilli
Signal transduction pathways for Umami, Sweet, Bitter, Sour, and Salty tastes
Taste Sensitivity and Adaptation:
Decrease in sensitivity with age
Rapid adaptation of taste sensation
Role of Saliva in Taste:
Dissolution of tastants to reach receptors
Washing away the stimulus
Taste Preferences and Aversions:
Mechanisms behind taste preference and aversion
Influence of receptors and neural pathways
Impact of Sensory Nerve Damage:
Degeneration of taste buds if the sensory nerve fiber is cut
Abnormalities of Taste Detection:
Conditions: Ageusia, Hypogeusia, Dysgeusia (parageusia)
Causes: Nerve damage, neurological disorders, infections, poor oral hygiene, adverse drug effects, deficiencies, aging, tobacco use, altered neurotransmitter levels
Neurotransmitters and Taste Threshold:
Effects of serotonin (5-HT) and norepinephrine (NE) on taste sensitivity
Supertasters:
25% of the population with heightened sensitivity to taste, especially bitterness
Increased number of fungiform papillae
KDM5 epigenetic modifiers as a focus for drug discovery
1. Epigenetic enzymes as drugable
targets for neurological disease
Christopher Wynder
PTM Discoveries
2. Epigenetics:so what
• Epigenetics is
responsible for the
variation seen in nature
amongst close relations
• Epigenetic regulation
also provides organisms
a mechanism to “tune”
gene expression based
environment (e.g. low
dietary fat and brain
development OR long
term injury-TBI, stroke)
Epigenetics also
explains how dogs
have so much variation
Twin studies show that many
differences in Autism
spectrum disorders seen
between siblings is likely
epigenetics
Or due to mutations in
epigenetic regulators
3. Epigenetic regulation has multiple jobs in the
brain
Stem cell
During development (and regeneration),
epigenetics plays a role in what kind of
neurons In the brain, Epigenetic
modifications are reused to modify
synaptic plasticity
Neurons have very few unique genes defining
their sub-type
Epigenetics modifies the levels of specific genes to
alter the physical parameters of the neurons.
4. Epigenetic definition
• “Non-genetic events” which result in stable
and inheritable gene expression patterns.
• Epigenetic changes are LONG term changes to
shape the portions of the genome that are
preferentially expressed
• Particularly relevant during development
• Regulation of histone modifications are a
significant and changeable example of an
epigenetic modification.
Epigenetic changes are about setting the
context for cell to nucleus signaling.
Limiting the response to any given stimuli
5. Epigenetics is the genome’s attempt at
grammar
• There are approx. 15 000 genes.
• Each one has a specific use and time when it should be used.
• In Epigenetics, is essentially how genes are organized and how
a cell decides which ones to use at a particular point in time.
• Histone modifications give context to allow words to be used
in the correct order.
• DNA methylation is used as the bookmarks to define chapters
(e.g. the “make a heart” or “respond to IGF”)
See Jane Stop make Run destroy brain Skinheart jeans workcells
See Jane Stop make Run destroy brain Skinheart jeans workcells
6. Maintaining expression of genes during
differentiation
• Gene regulation is divided into 2 sub-regions, regulatory and
promoter, and the transcribed region (gene encoding)
• In general, histone modification of the ORF is a on/off mark due to
the absolute requirement of these marks for RNA Polymerase read
through rate
• Histone modifications in the promoter generally work by modifying
RPol’s access
• Histone modifications in the regulatory region(s) are the “tuner”
increasing or decreasing of DNA binding transcription factors to
their elements
7. jmjN Domain - possible adaptor for protein-protein interaction
A/T Rich interaction Domain - DNA binding
PHD Domain (zinc finger similar to RING and FVYE domains)- Chromatin binding a triplicate of PHDs in KDM5s
jmjC histone demethylase - Catalytic domain a-ketoglutaric dehydrogenase
Zn finger Domain - C5H2 zinc finger DNA binding domain (chromatin binding)
KDM5b
KDM5c/d
65% Homology
1 1544
1535/
1516
KDM5 family structure
Wynder, C; Doughty, M; and Stalker, L. Epigenomics, June 2010
8. Neuron
During neural differentiation, neurons must
establish communication.
Tu parle
francais?
Do you
speak
english?
Yes
Cells use KDM5 family members to tune everything from the receptors and synaptic
shuttling machinery to the metabolic machinery.
This comprehensive control allows the neurons to control both short term plasticity and
long term “gene memory”
11. Recycling of cell cycle genes during neural
differentiation
KDM5b & c
targets
Frank and Tsai, Neuron 62 (2009)
12. Step-wise acclimation of histone modifications
regulate neural differentiation
3meH3K4
RPolIII
Low read through rate, low amounts
of mRNA made, therefore low
amounts protein, allows the cell to
block the expression of cell lineage
genes without 2nd signal.
Extremely low to no read through.
3meH3K27
HDM: KDM5(s)
HMT:KMT6
HDM: KDM6(s)
HMT:KMT2(s)
KDM5b (aka JARID1b/PLU1)
Adapted from Shilatifard Ann.Biochem 2006
13. Regulating the regulators
Apoptosis
Pro-neural
Pro-self renewal
Pro-neural
self renewal
Stage specific:
Seq. specific TFs:
Oct4, Sox2, FoxD3 NeuroD2, Sox1, FoxG1 Sox17, NGN2 Sox1, FoxD3
Pluripotent
Stem Cell
Neural
Progenitor
Differentiated
Neuron
Neural
Stem Cell
Ubiquitous factors
Chromatin/histone
Regulators:
KDM5b, Ring6a KDM5b, KDM5c, Ring6a KDM5c, Ring6a KDM5b, Ring6a
How do you control Ubiquitous factors
to modulate specific events?
14. Conserved mechanisms
KDM5
Moshkin et al, Mol. Cell 2010
In Drosophila KDM5
Is localized by interaction of
the NAP1-PF1 complex with
Gro-CtBP
NAP1 is a H2B-H2A dimer
Binding protein
Thought to be a
chaperone protein
15. In vitro
1. Recombinant proteins
2. Immunopurified complexes from ESCs (or tissue)
Testing epigenetic mechanisms in stem
cells
rKDM5b
Enzyme assaysWB from Nuc. Extracts mESCs
+
In vivo
1. Harvest spheres for RT-PCR and ChIP
2. Functional assay based cell markers(proteins)
Differentiation assays
Stalker L and Wynder C. Chapter 27. Methods in Molecular Biology. 2012
16. TLE4
H3
H4 H2A
H2B
K4 K43
H3
H4 H2A
H2B
K4
me
me
me
K43
me
me
Stem Cell
Cell cycle
inhibitor
RERE
Pluripotent
Self-renewing, proliferative
KDM5b
H3
H4 H2A
H2B
K4
me
me
me
K43
H3
H4 H2A
H2B
K4
me
me
me
K43
Multipotent
Proliferative
e.g. Neural stem cell
Stem Cell
Cell cycle
inhibitor
KDM5b
?
KDM5b
Lineage Committed
e.g. neuron
H3
H4 H2A
H2B
K4
me
me
me
K43
me
me
Cell cycle
inhibitor
TLE4
H3
H4 H2A
H2B
K4 K43
Stem Cell
RERE
KDM5b
KDM5b recruitment and activation
26. Linking biochemistry to biological
properties-Future directions
1. Define relationship between KDM5 and
cell signaling (MS sequencing and
verification)
2. Define the cell biology that is altered by
modulation of this system (Post injury
NSCs and iPSCs [NSCs v skin])
3. What is the role of these proteins in
both injury and recovery (mouse
models)
29. Anti YFP
(TLE4)
FLAG RERE (FL)
FLAG-RERE
Yfp
FLAG-RERE
YfpKDM5b
FLAG-RERE
YfpTLE4
FLAG-RERE
alone
Anti KDM5b
IP:FLAG
RERE regulates KDM5b interactions
and localization
FLAG-RERE
YFP-KDM5b
YFP-TLE4
YFPC1
Whole cell Extract
Chromatin Pellet
antiKDM5b
antiKDM5b
antiGAPDH
antiH3
30. Linking biochemistry to biological
properties-Future directions
1. Define relationship between KDM5 and
cell signaling (MS sequencing and
verification)
2. Define the cell biology that is altered by
modulation of this system (Post injury
NSCs and iPSCs [NSCs v skin])
3. What is the role of these proteins in
both injury and recovery (mouse
models)
31. Why iPSCs?
• Stem cells have the ability to differentiate into all cell lineages and self renew
• Recent advances have allowed us to convert skin into stem cells
• Since Rett syndrome mutations happen in all cells we can take skin and make stem cells by
epigenetically re-programming the skin
• iPS can be used as a model system to monitor neural differentiation to test where the errors are
and possibly what effect therapeutics have on these errors
Endoderm
Mesoderm
Ectoderm
Neurons
RBC
Skeletal system
Pancreatic
cells
Skin cells
Stem cell
Stomach cell
32. Harvest spheres for RT-PCR and ChIP
Represents “Day 1” of neurodifferentiation assay
Harvest Day 3
Harvest Day 14
Harvest Day 10
Harvest Day 8
Harvest Day 6
Harvest Day 5
Harvest Day 4
Mainly
Neural Stem
Completely
Differentiated
mESCs or iPSCs
Testing epigenetic mechanisms
1
2
Abrogation of KDM5/Co-factor here to elucidate the role
of this complex in acquisition of neural lineage.
Epigenetics of differentiation, can transient expression
block/enhance terminal differentiation
(can use adult sphere forming cells including Breast, Prostate from human/mouse)
3
Cell lineage selection; can expression during terminal
differentiation, modify the type of neuron that is made
OR
Cause de-differentiation/proliferation (iPSC/Cancer)
33. K4
me
K4
me
me
me
me
me
me
me
me
me
KDM5
TLE4
TSS
Defining the epigenetic mechanism of
neural specification
1. How does recruiters
choose the which KDM5
2. Is the interaction
antagonistic
3. How general is this
model i.e. how many
other recruiters are
there?
1. How is TLE4
recruited to KDM5
loci?
2. What is the role of
Post-translation
modifications in
TLE4 localization
Syn Ab/CcNSP construct
35. TLE4 function is required for neural
differentiation
TUJ1
CCNSP
Sox1
CCNSP
Transfection of the CCNSP Dom-Neg construct blocks both
early neural markers(Sox1) and late markers (TUJ1)
36. b c
feEYFP
EYFP
TLE4 CcNSP
TLE4 CcNSP
mCerulean TLE4 CcNSP
h iSox1
EYFP
Sox1
TLE4
Sox1
CcNSP
20 um
50 um
50 um
a
d
Day 4
Day 7
g
Day 2
0
0.5
1
1.5
2
Day Four Day Seven
Control
TLE4
CcNSP
NeuroD2 Expression
FoldChangeinExpression
Comparedtocontrol
j
**
** **
37. Understanding how KDM5 activity is
integrated into cell function
C.C. Inhibitors Differentiation Cell lineage
GSK3
/Erk BHC80
Ring6a
KMTx
K43
OR
TLE4
TLE4
Block interaction
Test affect during
neural diff.
(Glial v Neuron)
Block interaction
Test affect during
neural diff.
(Glial v Neuron)
38. No peptide TAT peptide TATH2B37-49
(K43me0)
mESCs
48hrspost
Neural diff (3 days) Neural diff (3 days) Neural diff (2 days)
Neuraldifferentiation
4-5dayspost
18 hrs after passage
48 hrs
Change to neural differentiation mediamESC media
NS NS NS
Day 0 Day 1 Day 3
48 hrs
Inhibitory peptide Gently remove intacted spheres by pipette
Change to ULB plates
40. Addition of H2BK43me0 peptides causes the formation of
neurospheres within 48 hours
NeuralFilamentb-TubulinIII
PhaseContrast
0
0.5
1
1.5
2
2.5
3
3.5
Control K43pep
B-TubIItranscription
*
N=3
41. Linking biochemistry to biological
properties-Future directions
1. Define relationship between KDM5 and
cell signaling (MS sequencing and
verification)
2. Define the cell biology that is altered by
modulation of this system (Post injury
NSCs and iPSCs [NSCs v skin])
3. What is the role of these proteins in
both injury and recovery (mouse
models)
42. Epigenetic modifications during
programming
• In mESCs H3K4 demethylases can block
differentiation when exogenously expressed.
• They act by repressing cell lineage factors.
• In non-stem cells (e.g HEK293 or HeLa) they can
activate transcription of stem cell factors (Oct4,
Sox2 and Nanog).
This means that altering KDM5 function in cells
can coax stem cells towards a preferred lineage.
43. Oct4 genes Sox2 genes KLF4 genes
KLF4
Taking advantage of signal dependent
epigenetic activity
In mESCs addition of Wnt3a mediates recruitment
of H3K4 demethylase(es) to
KLF4 target genes
Wnt3a
GSK3
??
44. KDM5b is sufficient for Nanog expression
in Skin cells
GFP+ Skin from mice
Transfected with KDM5b
Add mESC media
Culture 10d.
Nanog
Nanog
46. Colony formation in skin.
• KDM5b induces iPS-like cells.
• This induction is dependent
on the extra-cellular factors
• In appropriate media cells
become arborized.
• KDM5b alone is not sufficient
Putative “iPS clone”
Pre-colony
Neural media
KDM5b transfected
47. Therapeutic neural conversions
• KDM5b has the modulate the expression of a
variety of its target genes.
• The upstream signaling defines which targets.
• The changes do not appear to be permanent
suggesting that the loss of cell lineage markers is
insufficient for re-programming.
This may be advantageous for actually
regenerative medicine for direct to neural
conversion. Limiting the chance of transformation
(cancer).
49. KDM5b mechanism
• KDM5b regulates neural stem cells and potentially activity of
neurons through its target genes.
• KDM5b is regulated through both its localization and the
components of its complex.
• KDM5b is up-regulated in key populations after brain injury
Future Directions/Questions
Is KDM5b localization and activity 2 separate signals?
How do the components of the KDM5b complex effect post-
natal NSCs (i.e. differentiation, survival)?
How does alteration of KDM5 activity (either positive or
negative) effect injury response of NSCs (in situ)
50. In vivo Model
• KDM5b is an a pluripotency and a survival factor.
• The choice appears to be based on upstream input, and
most likely cell type (i.e. stem cell vs. committed).
• Taking advantage of this may allow for interrogation of
both long term injury and separately neural
differentiation.
1-7 days
RT-PCR
ChIP
Inject cells
sub-dermal/IM or
intra-cranial
Skin or other
tissue from
GFP+ mouse
Sol. factors
Neural Blood and skin
51. Acknowledgements
Leanne Stalker
Bijan Dey
Sean Keating
Ramin Shiekhattar
Joyce Papadimitriou-Taylor
Jonathan Bramson
Willa Liao
Ajapal Bhangu
Martin Doughty
Marc Meneghini
Ray Truant
Lise Munsie
Shawn Li
Wendy Zhu
Marek Galka
Huadong Liu