Have you considered that protein over-expression or inefficient mRNA knockdown may be masking physiological effects in your assays? Increasingly scientists are moving to endogenous gene-editing to characterise the function of their genes of interest.
Dr Chris Thorne from Cambridge Biotech Horizon Discovery discusses the ground breaking gene-editing technology CRISPR. The simplicity of experimental design has led to rapid adoption of the technology across the scientific community. However, challenges remain.
This Slidedeck focuses specifically on implementing CRISPR experiments, and explore a number of key considerations crucial to maximising chances of targeting success, whether your goal is to generate a knock-out or a knock-in. Chris also takes a look at some of the alternative uses of CRISPR, including sgRNA genome wide synthetic lethality screens.
The slides aim to support those researchers either planning to or already using CRISPR gene-editing in their lab. Horizon Discovery have also recently launched a program aimed specifically at academic cell biologists to promote the adoption of CRISPR by offering FREE CRISPR Reagents for knock-out cell line generation - more information available here. http://www.horizondiscovery.com/what-we-do/discovery-toolbox/genassist-crispr--raav-genome-editing-tools
Genome editing with the CRISPR-Cas9 system has become one of the major tools in modern biotechnology. This slide share discusses the fundamentals in a simple, easy to understand format.
Have you considered that protein over-expression or inefficient mRNA knockdown may be masking physiological effects in your assays? Increasingly scientists are moving to endogenous gene-editing to characterise the function of their genes of interest.
Dr Chris Thorne from Cambridge Biotech Horizon Discovery discusses the ground breaking gene-editing technology CRISPR. The simplicity of experimental design has led to rapid adoption of the technology across the scientific community. However, challenges remain.
This Slidedeck focuses specifically on implementing CRISPR experiments, and explore a number of key considerations crucial to maximising chances of targeting success, whether your goal is to generate a knock-out or a knock-in. Chris also takes a look at some of the alternative uses of CRISPR, including sgRNA genome wide synthetic lethality screens.
The slides aim to support those researchers either planning to or already using CRISPR gene-editing in their lab. Horizon Discovery have also recently launched a program aimed specifically at academic cell biologists to promote the adoption of CRISPR by offering FREE CRISPR Reagents for knock-out cell line generation - more information available here. http://www.horizondiscovery.com/what-we-do/discovery-toolbox/genassist-crispr--raav-genome-editing-tools
Genome editing with the CRISPR-Cas9 system has become one of the major tools in modern biotechnology. This slide share discusses the fundamentals in a simple, easy to understand format.
CRISPR-Cas9 is a genome editing tool that is creating a buzz in the science world. It is faster, cheaper and more accurate than previous techniques of editing DNA and has a wide range of potential applications.
i explained about basics of genome engineering and crispr system.
CRISPR will change the world and it is just the beginning, are you ready to meet the future? you think its great and beautiful or.....?
please give your feedback to my email
pooyanaghshbandi@yahoo.com
i am starting to write a critical and fantastic review article about CRISPR, if you are interested to join please contact me.
A simple version of the CRISPR/Cas system, CRISPR/Cas9, has been modified to edit genomes. By delivering the Cas9 nuclease complexed with a synthetic guide RNA (gRNA) into a cell, the cell's genome can be cut at a desired location, allowing existing genes to be removed and/or new ones added.
CRISPR (clustered regularly interspaced short palindromic repeats) is a family of DNA sequences found within the genomes of prokaryotic organisms such as bacteria and archaea. These sequences are derived from DNA fragments of bacteriophages that have previously infected the prokaryote and are used to detect and destroy DNA from similar phages during subsequent infections. Hence these sequences play a key role in the antiviral defense system of prokaryotes.
Cas9 (CRISPR-associated protein 9) is an enzyme that uses CRISPR sequences as a guide to recognize and cleave specific strands of DNA that are complementary to the CRISPR sequence. Cas9 enzymes together with CRISPR sequences form the basis of a technology known as CRISPR-Cas9 that can be used to edit genes within organisms.This editing process has a wide variety of applications including basic biological research, development of biotechnology products, and treatment of diseases.
The CRISPR-Cas system is a prokaryotic immune system that confers resistance to foreign genetic elements such as those present within plasmids and phages that provides a form of acquired immunity. RNA harboring the spacer sequence helps Cas (CRISPR-associated) proteins recognize and cut foreign pathogenic DNA. Other RNA-guided Cas proteins cut foreign RNA. CRISPR are found in approximately 50% of sequenced bacterial genomes and nearly 90% of sequenced archaea.
Genome editing is a method of making specific changes to the DNA of a cell or organism. An enzyme scissors the DNA at a specific sequence, and when this is repaired by the cell, a change or ‘edit’ is made to the sequence.
https://www.creative-biolabs.com/gene-therapy/approaches-to-genome-editing.htm
CRISPR is easily the best gene editing tool to date. For decades, scientists have been looking for a way to to perform precise changes to genetic sequences. In the past several years, researchers were able to exploit the immune systems of bacteria to edit the genome of other living cells. CRISPR is reported to have higher targeting efficiencies when compared to TALENs and Zinc Fingers. It is efficient, easy to use and cheap; making it a scalable genetic engineering tool that is highly desirable in various industry-wide applications.
It is very fast and new technique for detection and degradation of viral DNA and it is so helpful for us to understand how to degraded viral DNA... what type of function naturally present in bacteria........ so its very excellent technique
An Introduction to Crispr Genome EditingChris Thorne
In this short presentation, I make a case for doing genome editing vs some of the approaches that have gone before, describe some of the tools available, and the focus on CRISPR-Cas9, what it is, where it's come from and how it works.
Pharmacology Forever ! has been set as a meeting in recognition of Frits Peters tremendous involvement in pharmacology. This presentation discusses latest drug development methods and is illustrated by exemple of new drugs and target in oncology.
CRISPR-Cas9 is a genome editing tool that is creating a buzz in the science world. It is faster, cheaper and more accurate than previous techniques of editing DNA and has a wide range of potential applications.
i explained about basics of genome engineering and crispr system.
CRISPR will change the world and it is just the beginning, are you ready to meet the future? you think its great and beautiful or.....?
please give your feedback to my email
pooyanaghshbandi@yahoo.com
i am starting to write a critical and fantastic review article about CRISPR, if you are interested to join please contact me.
A simple version of the CRISPR/Cas system, CRISPR/Cas9, has been modified to edit genomes. By delivering the Cas9 nuclease complexed with a synthetic guide RNA (gRNA) into a cell, the cell's genome can be cut at a desired location, allowing existing genes to be removed and/or new ones added.
CRISPR (clustered regularly interspaced short palindromic repeats) is a family of DNA sequences found within the genomes of prokaryotic organisms such as bacteria and archaea. These sequences are derived from DNA fragments of bacteriophages that have previously infected the prokaryote and are used to detect and destroy DNA from similar phages during subsequent infections. Hence these sequences play a key role in the antiviral defense system of prokaryotes.
Cas9 (CRISPR-associated protein 9) is an enzyme that uses CRISPR sequences as a guide to recognize and cleave specific strands of DNA that are complementary to the CRISPR sequence. Cas9 enzymes together with CRISPR sequences form the basis of a technology known as CRISPR-Cas9 that can be used to edit genes within organisms.This editing process has a wide variety of applications including basic biological research, development of biotechnology products, and treatment of diseases.
The CRISPR-Cas system is a prokaryotic immune system that confers resistance to foreign genetic elements such as those present within plasmids and phages that provides a form of acquired immunity. RNA harboring the spacer sequence helps Cas (CRISPR-associated) proteins recognize and cut foreign pathogenic DNA. Other RNA-guided Cas proteins cut foreign RNA. CRISPR are found in approximately 50% of sequenced bacterial genomes and nearly 90% of sequenced archaea.
Genome editing is a method of making specific changes to the DNA of a cell or organism. An enzyme scissors the DNA at a specific sequence, and when this is repaired by the cell, a change or ‘edit’ is made to the sequence.
https://www.creative-biolabs.com/gene-therapy/approaches-to-genome-editing.htm
CRISPR is easily the best gene editing tool to date. For decades, scientists have been looking for a way to to perform precise changes to genetic sequences. In the past several years, researchers were able to exploit the immune systems of bacteria to edit the genome of other living cells. CRISPR is reported to have higher targeting efficiencies when compared to TALENs and Zinc Fingers. It is efficient, easy to use and cheap; making it a scalable genetic engineering tool that is highly desirable in various industry-wide applications.
It is very fast and new technique for detection and degradation of viral DNA and it is so helpful for us to understand how to degraded viral DNA... what type of function naturally present in bacteria........ so its very excellent technique
An Introduction to Crispr Genome EditingChris Thorne
In this short presentation, I make a case for doing genome editing vs some of the approaches that have gone before, describe some of the tools available, and the focus on CRISPR-Cas9, what it is, where it's come from and how it works.
Pharmacology Forever ! has been set as a meeting in recognition of Frits Peters tremendous involvement in pharmacology. This presentation discusses latest drug development methods and is illustrated by exemple of new drugs and target in oncology.
We can aid decision making from the pre-clinical to the clinical setting, supporting line of sight to the clinic, by identifying and translating crucial biomarker approaches into the real world.
INTRODUCTION
DNA VACCINES
GENE THERAPY
TIME LINE OF DEVELOPING GENE THERAPY
GENE THERAPY STRATEGIES
TECHNOLOGY OF CLASSICAL GENE THERAPY
PRINCIPLES OF GENE TRANSFER
VECTORS
VIRAL VECTORS
NON-VIRAL VECTORS
APPLICATIONS OF GENE THERAPY
ETHICAL IMPLICATIONS
THE FUTURE
CONCLUSION
REFERENCES
Gene therapy advanced treatments for a new era aranca special reportAranca
Aranca's Report on Gene Therapy - a promising tool for Cancer, Parkinson's, HIV, severe combined immuno-deficiencies, hemophilia etc. In this report, you will discover the challenges associated with Gene Therapy as well as its expected future.
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This presentation will highlight the promising new therapeutic strategies in the treatment of gliomas, with a focus on trials or therapies that will soon be available for Canadian patients.
View the YouTube video: https://youtu.be/ibbEuvSF7xY
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The dream of any physician and consequently every patient is to receive the right treatment in the right time with cost effectiveness. To achieve this goal, the 3 pillars: evidence based medicine, clinical research innovation & resources utilization should be integrated efficiently.
In this presentation, I'll try to comprehensively review the following:
1- How are we used to perform clinical trials in Oncology?
2- Does it fits in today’s needs?
3- Integration of biology knowledge in shaping drug development
4- New Clinical trial designs “Can they offer solution for accelerating drug development?”
5- The supporting infrastructure role in clinical trial execution
Our fifth webinar in the MDC Connects Series 2021 | A Guide to Complex Medicines.
This slide deck takes a closer look at physicochemical characterisation new and novel approaches to understand the pharmacokinetics of complex drugs.
Juliana Maynard (MDC)
Wielding the Double-Edge Sword of Cardiac Biomarkers in Clinical Trials: A Di...Medpace
Learn best practices for utilizing cardiac biomarkers across various components of a clinical trial from Dr. James Januzzi, a leading expert in cardiovascular biomarkers.
Avoiding Common Pitfalls in Cell and Gene Therapy TrialsMedpace
Our experts will dive into case studies explaining the issues that arose with past projects, and how these studies got back on track. The knowledge gained from these experiences accelerate CGT development by avoiding potential pitfalls and getting ahead of regulatory and operational issues. Our team will walk you through the delays associated with these pitfalls and how to avoid them.
Considerations for the Next Wave of COVID-19 DevelopmentMedpace
What did we learn from the first wave of COVID-specific studies that can be applied to those studies that will follow? Medpace has gathered some of its medical and operational team experts who have been on the front-line in designing and conducting trials for SARS-CoV-2 to answer some frequently asked questions, trends they see emerging and considerations for future development.
COVID-19 Product Development and Clinical Trials: Considerations from Europea...Medpace
Join experts from Medpace’s regulatory and operational teams in this webinar as they provide insights and considerations on how to accelerate product development for COVID-19 during different stages
Part 3: Rare Disease Clinical Development – Strategies for Ensuring Endpoint ...Medpace
n this free webinar, Medpace partners with Michelle Eagle of ATOM International, a provider of CE training for clinical trials across the world, to discuss approaches and steps that can be taken to ensure quality and integrity.
Getting Ahead of the Evolving Landscape in RadiopharmaceuticalsMedpace
In this webinar devoted to radiopharmaceuticals, the featured speakers will explore the scientific, operational and regulatory considerations for radiopharmaceuticals. With a focus on oncology, they will discuss the current regulatory landscape and how this impacts overall development programs. The speakers will explore the challenges of conducting radiopharmaceutical trials, offering insights into trial start-up, site selection and operational aspects to seamlessly execute these studies as part of clinical development plans.
Challenges and Considerations in Clinical Development of "Targeted Therapies"...Medpace
In this webinar, Medpace experts discuss key clinical, operational and laboratory considerations, lessons-learned, and best practices for accelerating the global development of safe and effective targeted therapeutics, using acute myeloid leukemia (AML) to highlight the complexities.
The RACE for Children Act Will Change the Landscape for Pediatric Cancer Rese...Medpace
In this webinar, we explore the regulatory implications of the RACE for Children Act and what this law means for your development program, particularly with navigating the change in requirements for pediatric oncology trials. Furthermore, we explore the challenges of executing oncology trials in pediatric populations and offer insight into design and operational aspects to seamlessly execute these studies as part of your clinical development plan
Identifying High Performing Sites and Engaging PatientsMedpace
One of the biggest challenges facing any clinical trial is how to identify the sites with the greatest potential to engage and retain patients. Applying decades of experience to the topic, Medpace experts will share considerations, lessons-learned and best practices for developing patient recruitment strategies to put you on the path for optimal success.
Challenges and Considerations in Designing and Conducting Immuno-Oncology Cli...Medpace
Given the accelerating pace of immuno-oncology clinical research, awareness of the specific challenges and considerations in designing and conducting successful trials for these new agents is critical.
Webinar: Oncology Trial Recruitment: Challenging Indications and Challenging ...Medpace
Medpace experts discuss how to overcome oncology recruitment challenges for clinical trials for specific populations, indications, and challenging studies.
What Happens After Your Device is Approved? Collecting Data in the Real WorldMedpace
In this workshop, Medpace will discuss key considerations for generating real-world evidence and how to apply critical insights in order to drive late-stage clinical research. To listen to this presentation, visit https://vimeo.com/168768256
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
Best Ayurvedic medicine for Gas and IndigestionSwastikAyurveda
Here is the updated list of Top Best Ayurvedic medicine for Gas and Indigestion and those are Gas-O-Go Syp for Dyspepsia | Lavizyme Syrup for Acidity | Yumzyme Hepatoprotective Capsules etc
Local Advanced Lung Cancer: Artificial Intelligence, Synergetics, Complex Sys...Oleg Kshivets
Overall life span (LS) was 1671.7±1721.6 days and cumulative 5YS reached 62.4%, 10 years – 50.4%, 20 years – 44.6%. 94 LCP lived more than 5 years without cancer (LS=2958.6±1723.6 days), 22 – more than 10 years (LS=5571±1841.8 days). 67 LCP died because of LC (LS=471.9±344 days). AT significantly improved 5YS (68% vs. 53.7%) (P=0.028 by log-rank test). Cox modeling displayed that 5YS of LCP significantly depended on: N0-N12, T3-4, blood cell circuit, cell ratio factors (ratio between cancer cells-CC and blood cells subpopulations), LC cell dynamics, recalcification time, heparin tolerance, prothrombin index, protein, AT, procedure type (P=0.000-0.031). Neural networks, genetic algorithm selection and bootstrap simulation revealed relationships between 5YS and N0-12 (rank=1), thrombocytes/CC (rank=2), segmented neutrophils/CC (3), eosinophils/CC (4), erythrocytes/CC (5), healthy cells/CC (6), lymphocytes/CC (7), stick neutrophils/CC (8), leucocytes/CC (9), monocytes/CC (10). Correct prediction of 5YS was 100% by neural networks computing (error=0.000; area under ROC curve=1.0).
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
Muktapishti is a traditional Ayurvedic preparation made from Shoditha Mukta (Purified Pearl), is believed to help regulate thyroid function and reduce symptoms of hyperthyroidism due to its cooling and balancing properties. Clinical evidence on its efficacy remains limited, necessitating further research to validate its therapeutic benefits.
New Drug Discovery and Development .....NEHA GUPTA
The "New Drug Discovery and Development" process involves the identification, design, testing, and manufacturing of novel pharmaceutical compounds with the aim of introducing new and improved treatments for various medical conditions. This comprehensive endeavor encompasses various stages, including target identification, preclinical studies, clinical trials, regulatory approval, and post-market surveillance. It involves multidisciplinary collaboration among scientists, researchers, clinicians, regulatory experts, and pharmaceutical companies to bring innovative therapies to market and address unmet medical needs.
Recomendações da OMS sobre cuidados maternos e neonatais para uma experiência pós-natal positiva.
Em consonância com os ODS – Objetivos do Desenvolvimento Sustentável e a Estratégia Global para a Saúde das Mulheres, Crianças e Adolescentes, e aplicando uma abordagem baseada nos direitos humanos, os esforços de cuidados pós-natais devem expandir-se para além da cobertura e da simples sobrevivência, de modo a incluir cuidados de qualidade.
Estas diretrizes visam melhorar a qualidade dos cuidados pós-natais essenciais e de rotina prestados às mulheres e aos recém-nascidos, com o objetivo final de melhorar a saúde e o bem-estar materno e neonatal.
Uma “experiência pós-natal positiva” é um resultado importante para todas as mulheres que dão à luz e para os seus recém-nascidos, estabelecendo as bases para a melhoria da saúde e do bem-estar a curto e longo prazo. Uma experiência pós-natal positiva é definida como aquela em que as mulheres, pessoas que gestam, os recém-nascidos, os casais, os pais, os cuidadores e as famílias recebem informação consistente, garantia e apoio de profissionais de saúde motivados; e onde um sistema de saúde flexível e com recursos reconheça as necessidades das mulheres e dos bebês e respeite o seu contexto cultural.
Estas diretrizes consolidadas apresentam algumas recomendações novas e já bem fundamentadas sobre cuidados pós-natais de rotina para mulheres e neonatos que recebem cuidados no pós-parto em unidades de saúde ou na comunidade, independentemente dos recursos disponíveis.
É fornecido um conjunto abrangente de recomendações para cuidados durante o período puerperal, com ênfase nos cuidados essenciais que todas as mulheres e recém-nascidos devem receber, e com a devida atenção à qualidade dos cuidados; isto é, a entrega e a experiência do cuidado recebido. Estas diretrizes atualizam e ampliam as recomendações da OMS de 2014 sobre cuidados pós-natais da mãe e do recém-nascido e complementam as atuais diretrizes da OMS sobre a gestão de complicações pós-natais.
O estabelecimento da amamentação e o manejo das principais intercorrências é contemplada.
Recomendamos muito.
Vamos discutir essas recomendações no nosso curso de pós-graduação em Aleitamento no Instituto Ciclos.
Esta publicação só está disponível em inglês até o momento.
Prof. Marcus Renato de Carvalho
www.agostodourado.com
- Video recording of this lecture in English language: https://youtu.be/lK81BzxMqdo
- Video recording of this lecture in Arabic language: https://youtu.be/Ve4P0COk9OI
- Link to download the book free: https://nephrotube.blogspot.com/p/nephrotube-nephrology-books.html
- Link to NephroTube website: www.NephroTube.com
- Link to NephroTube social media accounts: https://nephrotube.blogspot.com/p/join-nephrotube-on-social-media.html
Flu Vaccine Alert in Bangalore Karnatakaaddon Scans
As flu season approaches, health officials in Bangalore, Karnataka, are urging residents to get their flu vaccinations. The seasonal flu, while common, can lead to severe health complications, particularly for vulnerable populations such as young children, the elderly, and those with underlying health conditions.
Dr. Vidisha Kumari, a leading epidemiologist in Bangalore, emphasizes the importance of getting vaccinated. "The flu vaccine is our best defense against the influenza virus. It not only protects individuals but also helps prevent the spread of the virus in our communities," he says.
This year, the flu season is expected to coincide with a potential increase in other respiratory illnesses. The Karnataka Health Department has launched an awareness campaign highlighting the significance of flu vaccinations. They have set up multiple vaccination centers across Bangalore, making it convenient for residents to receive their shots.
To encourage widespread vaccination, the government is also collaborating with local schools, workplaces, and community centers to facilitate vaccination drives. Special attention is being given to ensuring that the vaccine is accessible to all, including marginalized communities who may have limited access to healthcare.
Residents are reminded that the flu vaccine is safe and effective. Common side effects are mild and may include soreness at the injection site, mild fever, or muscle aches. These side effects are generally short-lived and far less severe than the flu itself.
Healthcare providers are also stressing the importance of continuing COVID-19 precautions. Wearing masks, practicing good hand hygiene, and maintaining social distancing are still crucial, especially in crowded places.
Protect yourself and your loved ones by getting vaccinated. Together, we can help keep Bangalore healthy and safe this flu season. For more information on vaccination centers and schedules, residents can visit the Karnataka Health Department’s official website or follow their social media pages.
Stay informed, stay safe, and get your flu shot today!
2. Genetic Engineering
2 Physician Led | Therapeutically Focused
“…the process of making targeted modifications to
the genome, its contexts (e.g., epigenetic marks),
or its outputs (e.g., transcripts)”
(Hsu et al, 2014).
4. Transcription Activator Like Effector Nuclease (TALEN):
Application in Duchenne Muscular Dystrophy
4 Physician Led | Therapeutically Focused
X Y
DMD gene
Xp21 • DMD gene: 79 exons, deletions,
duplications or loss can lead to lack of
functional dystrophin protein
• Large size renders traditional AAV
based gene editing difficult
• Li et al. (2015) used TALEN to correct
in iPSCs via exon knockin and
demonstrated proof of principle
• TALEN can have off target
mutagenesis
Li et al, Stem Cell 2015
5. CRISPR: Clustered Regularly Interspaced
Short Palindromic Repeats
5 Physician Led | Therapeutically Focused
o First described in E. coli and determined to be part of
the bacterial innate immune system versus
bacteriophages
o Consists of short segments of DNA that are palindromes
interspaced with spacer DNA
o The spacer DNA is identical in sequence to viral
(bacteriophage) DNA
o There are additional CRISPR associated proteins:
cas proteins that are typically helicases or nucleases
Spacer
DNA
cas Spacer
DNA
Spacer
DNA
Spacer
DNA
6. CRISPR Basics
Physician Led | Therapeutically Focused6 Physician Led | Therapeutically Focused6
Spacer
DNA
cas Spacer
DNA
Spacer
DNA
Spacer
DNA
Bacteria Cell Wall
7. cas Protein and crRNA Produced
Physician Led | Therapeutically Focused7 Physician Led | Therapeutically Focused7
Spacer
DNA
cas Spacer
DNA
Spacer
DNA
Spacer
DNA
cas protein /crRNA
complex
Bacteria Cell Wall
8. Physician Led | Therapeutically Focused8 Physician Led | Therapeutically Focused8
Spacer
DNA
cas Spacer
DNA
Spacer
DNA
Spacer
DNA
cas Protein
A New Bacteriophage Arrives
Bacteria Cell Wall
9. Bacteriophage Denied!!
New Spacer DNA Incorporated into Bacterial Genome for Next Time…
Physician Led | Therapeutically Focused9 Physician Led | Therapeutically Focused9
New
Spacer
DNA
Spacer
DNA
cas Spacer
DNA
Spacer
DNA
Spacer
DNA
Bacteria Cell Wall
10. Physician Led | Therapeutically Focused10
The Breakthrough
Jinek et al, Science, 2012
11. Physician Led | Therapeutically Focused11
Next?
Physician Led | Therapeutically Focused
o Transcription
interference
o Co-activation and
interference
o Transcription
activation
o Epigenetic modifiers
11
Dominguez et al., Nature Reviews: Molecular Cell Biology, 17, 5-15, Jan 2016
12. Clinical Human Applications of CRISPR
12 Physician Led | Therapeutically Focused
o Viral infections: HIV, HSV, VZV
Inserted viral genome could be removed
by altering immune cells
Human embryos: Kang and colleagues
inserted the CCR5Δ32 allele into early
human 3PN embryos
o Cancer
Mutation driven cancers
Kang, X. et al. J Assist Reprod Genet 33, 581, 2016
13. Clinical Human Applications of CRISPR?
13 Physician Led | Therapeutically Focused
o Genetic diseases
Remove or add the sequence
that is causing the disease
o Transplantation
Gene editing of mismatched human
or even non-human mammals as
potential organ donors
Editing will reduce risk of immune
responses and rejection when using
mismatched organs/tissues/cells
14. Human Experience
14 Physician Led | Therapeutically Focused
o Liang et al using human
tripronuclear zygotes cleaved the
HBB gene with a CRISPR/Cas9-
mediated system
Low efficiency and edited
embryos were mosaic with off
target cleavage
o Other clinical trials forthcoming:
Editas
CRISPR Therapeutics
Caribou Biosciences
Intellia Therapeutics
Liang et al, Protein and Cell 2015
16. The NIH Recombinant Advisory
Committee has Approved the First US Trial
16 Physician Led | Therapeutically Focused
The University of
Pennsylvania:
combination of PD-1 and
NY-ESO-1 and LAGE-1
in human cancer
Time, 2016
17. Limitless Applications…
Physician Led | Therapeutically Focused17
• Drug development – optimize
biotech manufacture
• Disease models
• Ecological vector control –
mosquito sterilization
• Biofuels
• Agriculture – modification of
crop strains or animals
19. Clinical Development Considerations for
Gene Editing Technology
19 Physician Led | Therapeutically Focused
o Therapies may provide life-long cure through a single
treatment
o CRISPR technology has made gene-editing much more
accessible and has broadened the range of targets
o Regulatory and ethical frameworks
o Bring new therapies to the clinic via a safe and rapid
pathway
20. Physician Led | Therapeutically Focused
o Most likely to be largest
area of clinical
development using
CRISPR technology
o Regulations available to
govern these applications
o No new ethical
concerns/issues
o Research permitted
o Therapies being
developed
o Considered for some
indications – would alter
the genome in all cells and
become heritable
o Ethical concerns
o Inconsistent and variable
guidelines and regulation
o Limited research permitted
o Development of therapies
may be restricted
Somatic Cell Therapy Germ Line Therapy
20
21. Ethical Concerns
o International Summit on Human Gene Editing (Dec 2015)
Concerns over germ line editing – need for an ongoing forum
o NAS/NAM Meeting (April 2016)
All aspects of human germline editing, consequences, regulation and potential
applications
Committee assembled to perform a year-long in-depth study
o EU CT Directive (2001/20/EC) does not allow germ line editing (Article 9)
o NIH RAC will not review proposals on germ line editing
o Eugenic practices prohibited:
Oviedo Convention
Convention on Human Rights and Biomedicine (1997)
Article 3(2) of the Charter of Fundamental Rights of EU prohibits eugenic practices
Non-Therapeutic Use / Enhancement
Physician Led | Therapeutically Focused21
22. Regulatory Challenges
o CRISPR/Cas-based gene editing of
somatic cell therapies will use in vivo or
ex vivo strategies
o Current regulations for gene therapy and
cell therapy will regulate CRISPR-based
therapies
o Regulators will need to stay up to date
with rapid technology advances
o Pathways to market will need to be
flexible and allow timely patient access
to therapies
Somatic Cell Therapies
Physician Led | Therapeutically Focused22
23. o Off-target effects/genotoxicity
Improvements in targeting of CRISPR/Cas9 system
• Methods to assess genome-wide off-target effects
• Need to ensure there is no detectable germ line modification
o Efficacy
Single administration may be sufficient but need to consider multiple
treatments
Control of CRISPR/Cas editing
o Animal models
Significant area for regulators and companies
Safety and Efficacy
Physician Led | Therapeutically Focused23
24. Physician Led | Therapeutically Focused
o Complex manufacturing processes
o Release testing
o Shelf-life may be short (hours)
o Stability and transportation logistics
are important considerations
Quality/Manufacturing and Administration
24
o Patients may be treated in
specialized centers
o Manufacture based at site of
administration
o Patients to be localized at these
sites – international travel for
treatment will become more
common
25. o Role of RAC in the US
Recent revisions to streamline the process
o Review of gene therapy studies in EU
Additional time for review, may involve expert committees
Use of modified viral vectors requires authorisation for use of GMOs
o Regulators gain experience with gene editing therapies (quality,
safety, efficacy)
Increased focus on review by ECs/IRBs may raise additional
questions and impact the start-up process for CTs
Clinical Trial Considerations
25 Physician Led | Therapeutically Focused
26. Regulatory Pathways to the Market
o EU, Japan and US recognize the importance of
faster transit through the current regulatory
pathways
EU – Adaptive pathways (conditional approvals),
PRIME (PRIority MEdicines), accelerated
assessment
Japan –PMD Act, expedited approval system for
regenerative medicine products
US – fast track, breakthrough therapy,
accelerated approval, priority review
Timely and Flexible; Faster Access to Developing Therapies
Physician Led | Therapeutically Focused26
27. Cost and Reimbursement
o CRISPR-based therapy that provides a one-off
lifetime cure will come at a high development
cost
Rare diseases will only have a small number
of potential patients
o Single high-cost therapy treatment could remove
a lifetime’s cost of existing treatment and be
more effective
The Market Challenge
Physician Led | Therapeutically Focused27
o Health Technology Assessment bodies (HTAs) may need to consider new
approaches to pricing
Strimvelis (GSK) - pay for performance agreement in Italy (AIFA)
28. Future Direction of Clinical Development
o CRISPR/Cas9-based gene editing of cells and tissues will be an
exciting and rapid area of development in the coming years
Anticipate that rare genetic diseases, mutation-driven malignancies
and cardiovascular indications will be key areas of development
Recognize that the potential is vast and applications may be limitless
o Flexible approach to the regulatory pathways is essential
Developers, ethicists and regulators should be discussing
collaboratively at an early stage and throughout the development
pathway
o Long term efficacy and safety will demonstrate the value of this
technology and demonstrate its potential to treat complex and
challenging diseases
Physician Led | Therapeutically Focused28
29. Physician Led | Therapeutically Focused29
“Genome editing holds great promise to provide a
precise set of tools for counteracting genetic
diseases.
But as Spiderman cautions, ‘With great power,
there must come great responsibility’
Moving these methods to clinical applications must
proceed judiciously….(and) under appropriate
regulatory oversight”
Kohn et al, Blood (2016, 127: 2553 - 2560)
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Good morning. Trevor and I would like to thank the audience for their attendance today. Together, Trevor and I will review the basics of gene editing, recent clinical experience with the technology followed by a discussion of the regulatory and ethical implications of human genetic engineering. We will have time for questions at the end.
Gene engineering has evolved over time from a method to generate genetic knock-in and knock-out animals to genetic surgery for human diseases
The quest for editing the human genome to treat disease has been an ongoing objective of human medicine for many years. The ability to form a precise DNA break, followed by editing or correction has been attempted via a variety of techniques: meganucleases, oligonucleotides, peptide nucleic acids and more recently zinc finger nucleases and transcription activator-like effector nucleases (TALEN). The most recent addition, clustered regularly interspaced short palindromic repeats (CRISPR) has created an increasing level of interest and scrutiny.
Like any advance in medicine, genetic engineering offers great promise and great responsibility.
I will focus the discussion today on three of the gene editing technologies: Zinc finger nuclease, TALEN and CRISPR.
In brief, zinc finger nucleases a class of engineered DNA-binding proteins that enable targeted double-strand breaks in DNA at user-selected locations. Each Zinc Finger Nuclease is comprised of a DNA binding domain and a DNA cleaving domain comprised of the nuclease domain of Fok I. When the DNA-binding and DNA-cleaving domains are fused together, a highly-specific pair of 'genomic scissors' are created. Zinc finger nucleases can recognize independently 3-4 DNA bases and when linked can target specific DNA sequences, bind and cleave and either through non homologous end joining or homologous recombination can either replace or mutate the target gene
This technology has been used in plant and mammalian cells. The NEJM article here provides an example of a human clinical trial that used ZFN.
Following a clinical observation of a less aggressive clinical course in select HIV infected patients who were heterozygous for the CCR5 delta 32 gene combined with the single report of undetectable HIV following an allogeneic cell transplant from a homozygous CCR5 delta 32 donor, a clinical trial was undertaken. the question was raised if you could infuse CD4 T cells that had undergone gene editing rendering them CCR5 delta 32 deficient.
12 patients enrolled in a phase 1 trial involving CCR5 modified CD4 T cells. One of four evaluable patients had undetectable HIV and the trial was felt to be safe with one related SAE.
Concerns regarding off target cleavage have been raised with this technology
A second gene editing technology has been used in clinical trials, TALEN.
TALEN: are engineered restriction enzymes that cut specific sequences of DNA. The basic construct consists of a transcription activator-like effectors (TALEs) that is bound to practically any desired DNA sequence, so when combined with a nuclease, a resulting specific DNA cut will occur.
Transcription activator-like effector nucleases (TALEN) were the next evolution of chimeric nucleases that are more readily engineered to specific binding domains providing more specificity. Based upon the discovery by Scholze and Boch and colleagues of plant pathogens, TALE nucleases represent a method to target endogenous genes in cells.
Clinical applications of TALEN have been published. In 2015, Li and colleagues published on TALEN correction in stem cell correction of the Duchenne muscular dystrophy gene. The DMD gene comprised on 79 exons, deletions and duplications that can lead to the lack of function dystrophin protein. The large size makes traditional adenovirus gene therapy difficult. In the publication, they demonstrated proof of principle.
TALEN technology was also reported in the UK with the use of TALEN in chimeric antigen receptor allogeneic modified T cells in pediatric pre B cell ALL where the TCR was edited out using TALEN technology.
The limitations of TALEN however are the off target effect
Now moving on to the next technology: Clustered regularly interspaced short palindromic repeats or CRISPR
CRISPR has been known by bacteriologists for many years and first described as an innate immune system used by bacteria to fend off viral infection via bacteriophages. It was noted that there were short segments of palindromic DNA interspaced with spacer DNA. The spacer DNA being unique and has been found to be identical to viral DNA. In addition to the DNA sequence, cas or CRISPR associated genes that encode for proteins are found that are typically helicases or nucleases. As CRISPR is a newer technology, we will take a moment to describe how it functions in bacterial cells.
Lets go through a very basic description of CRISPR in bacteria. A bacteria is infected by a bacteriophage, the virus injects viral DNA into the bacteria.
If the bacteria has seen this viral DNA before, the cas protein is transcribed along with transcription crRNA which fits into the cas protein complex and using the helicases and nucleases of the cas protein complex to break apart the viral DNA
What if there is a novel viral DNA introduced, the CRISPR system will generate a new class 1 cas protein will break apart the viral DNA
And incorporate the new spacer DNA into the bacterial genome. When infected by the bacteriophage in the future, the bacteria would have adaptive immunity to that virus and use the CRISPR cas system to degrade the DNA. This system was known for some time as an immune system of bacteria. The innovation occurred when several investigators evaluated one CRISPR system in Strep pyogenes.
In 2012, using Strep pyogenes, Doudna, Charpentier and others described a modified mechanism to use the CRISPR system with the strep pyogenes cas proteins: cas 9. The paper published in Science in 2012, was a revolutionary change in DNA editing as it provided an elegant system that could be used to precisely edit DNA.
In the native system, cas9 is a nuclease. There are two RNA’s formed: the crRNA and an additional RNA, the tracrRNA which holds the crRNA in place.
The advance described in the paper was the creation of a chimera of the entire system that combined the tracrRNA and crRNA into one guide RNA: the gRNA. Thus the system created is the cas9 protein and the gRNA, the chimera. The system would work very similarly to just described in the simple diagrams in preceding slides.
It works by taking the sequence of DNA that you want to edit and creating a gRNA with that exact sequence in the gRNA. Then insert the chimera into the target cell, the cas9 will cut the DNA at the exact sequence. Then cell will direct endogenous repair mechanisms within the cell to repair the cut the DNA either without target sequence of DNA
Alternatively, you can insert a gene, in this instance you have the cas9, the gRNA and the host RNA. The system will cut the DNA at the target based on the gRNA, then the host RNA will be inserted into the DNA as a new segment.
What was striking about this advance was the elegant simplicity of the method and the explosion of applications that commenced following the publication was astounding.
With these observations, a flurry of CRISPR activity has commenced. In Dominguez Nature Reviews, there are elegant descriptions of mechanisms to interfere with transcription of a DNA which one could consider in an overexpression state, for example p53. There are applications of co-activation and interference and finally transcription activation
In addition to these direct DNA applications, scientists are investigating targeting the epigenome using the CRISPR system.
Certainly, we can all contemplate the myriad of applications that gene editing could be considered. We have discussed the ZFN approach to HIV infection and Kang and colleagues reported in April of this year with the use of the CRISPR cas 9 system to insert the HIV resistance gene CCR5 into human embryos. The TALEN approach to T cell receptors which one could surmise would be possible fields of clinical investigation.
Additionally, as we described TALEN technology application in Duchennes, one could consider removal or addition of genetic material as it applies to genetic diseases. Finally, the field of human organ or stem cell transplantation in theory could be a possible application of CRISPR/cas even to the point of using mismatched human or non-human donors which would revolutionize human transplantation.
Clinical reports are beginning to be published using the CRISPR cas 9 system. In 2015, Liang and colleagues reported on human zygotes where the human beta globin gene was cleaved with the system. They demonstrated low efficiency and off target cleavage but did demonstrate proof of principle.
Additional human trials are forthcoming with several companies and academic centers developing CRISPR based systems for human diseases.
A quick check of the clinicaltrials.gov web page found 4 clinical trials in China that as of Sept 2016 were not recruiting patients but were targeting a variety of human malignancies. As a clinical research organization, it is astounding the speed at which the CRISPR/cas system has gone from bench to bedside and the global reach of this technology.
And in the US, the University of Pennsylvania received RAC approval for the first US trial using CRISPR/cas technology targeting a combination of PD-1, NY-ESO1 and LAGE-1 in human malignancy.
Gene editing technology has limitless applications, from drug development, disease models, ecological vector control, biofuels and most recently reported in Sweden modification of crops, in this case a CRISPR modified cabbage and most recently a CRISPR modified cucumber was generated and eaten this month.
In this quick review of gene editing technology, I have hopefully touched on a variety of gene editing technologies, given a simple explanation of the newest addition, CRISPR and would like to share the screen with my colleague Trevor Walker who will discuss the regulatory and ethical implications of these technologies.
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