The Electrophoretic Mobility Shift Assay (EMSA) is used to detect protein complexes with nucleic acids such as DNA and RNA. It involves incubating a purified protein or cell extract with a radioactively labeled probe, then separating bound and unbound complexes on a gel during electrophoresis. The protein-nucleic acid complexes migrate more slowly than unbound probes. The assay can identify sequence-specific DNA and RNA binding proteins and determine binding specificity using competitive experiments with specific and nonspecific fragments. It is a basic method to study DNA-protein and RNA-protein interactions.
This is technique used widely for protein separation from a mixture and is very easy and less costly method. Slides cover all essential points about EMSA and it is quite interesting to know that how it detect and separate different proteins and their mobility shift assay.
''Electrophoretic Mobility Shift Assay'' by KATE, Wisdom DeebekeWisdom Deebeke Kate
This assessed presentation was delivered by me, together with other three course mates. The aim of the presentation was to describe the basic principles, methods involved in EMSA, and some of its application in molecular biology to study the interactions between proteins and DNA. Delivered on 9th December, 2013 with Lolomari Songo, Nicholas Leach & Abhay Jethwani.
This is technique used widely for protein separation from a mixture and is very easy and less costly method. Slides cover all essential points about EMSA and it is quite interesting to know that how it detect and separate different proteins and their mobility shift assay.
''Electrophoretic Mobility Shift Assay'' by KATE, Wisdom DeebekeWisdom Deebeke Kate
This assessed presentation was delivered by me, together with other three course mates. The aim of the presentation was to describe the basic principles, methods involved in EMSA, and some of its application in molecular biology to study the interactions between proteins and DNA. Delivered on 9th December, 2013 with Lolomari Songo, Nicholas Leach & Abhay Jethwani.
This presentation contains information about restriction enzymes, its nomenclature, restriction digestion, and its application. This also contains information about the chemicals used in restriction and also explains the general procedure of restriction digestion of DNA
Transcriptomics is the study of RNA, single-stranded nucleic acid, which was not separated from the DNA world until the central dogma was formulated by Francis Crick in 1958, i.e., the idea that genetic information is transcribed from DNA to RNA and then translated from RNA into protein.
BAC & YAC are artificially prepared chromosomes to clone DNA sequences.yeast artificial chromosome is capable of carrying upto 1000 kbp of inserted DNA sequence
Creation of a cDNA library starts with mRNA instead of DNA. Messenger RNA carries encoded information from DNA to ribosomes for translation into protein. To create a cDNA library, these mRNA molecules are treated with the enzyme reverse transcriptase, which is used to make a DNA copy of an mRNA (i.e., cDNA). A cDNA library represents a sampling of the transcribed genes, but a genomic library includes untranscribed regions.
To modifying the structure of a specific gene.
Gene targeting vector introduced into the cell.
Vector modifies the normal chromosomal gene through homologous recombination.
Useful in treating some human genetic disorders – Hemophilia, Duchenne Muscular Dystrophy.
Treating human diseases by genetic approaches – Gene Therapy.
Gene Therapy – Replacing the defective gene by normal copy of the gene.
Expressed sequence tag/EST is a short partial sequence, typically 200-400 bp long, of a complimentary DNA/Cdna.
EST is a short sub-sequence of a cDNA sequence.
Used to identify gene transcripts, and are instrumental in gene discovery and in gene-sequence determination.
Approximately 74.2 million ESTs are available in public databases.
EST results from one-short sequencing of a cloned cDNA.
Low-quality fragments.
Length is approximately 500 to 800 nucleotides.
This presentation contains information about restriction enzymes, its nomenclature, restriction digestion, and its application. This also contains information about the chemicals used in restriction and also explains the general procedure of restriction digestion of DNA
Transcriptomics is the study of RNA, single-stranded nucleic acid, which was not separated from the DNA world until the central dogma was formulated by Francis Crick in 1958, i.e., the idea that genetic information is transcribed from DNA to RNA and then translated from RNA into protein.
BAC & YAC are artificially prepared chromosomes to clone DNA sequences.yeast artificial chromosome is capable of carrying upto 1000 kbp of inserted DNA sequence
Creation of a cDNA library starts with mRNA instead of DNA. Messenger RNA carries encoded information from DNA to ribosomes for translation into protein. To create a cDNA library, these mRNA molecules are treated with the enzyme reverse transcriptase, which is used to make a DNA copy of an mRNA (i.e., cDNA). A cDNA library represents a sampling of the transcribed genes, but a genomic library includes untranscribed regions.
To modifying the structure of a specific gene.
Gene targeting vector introduced into the cell.
Vector modifies the normal chromosomal gene through homologous recombination.
Useful in treating some human genetic disorders – Hemophilia, Duchenne Muscular Dystrophy.
Treating human diseases by genetic approaches – Gene Therapy.
Gene Therapy – Replacing the defective gene by normal copy of the gene.
Expressed sequence tag/EST is a short partial sequence, typically 200-400 bp long, of a complimentary DNA/Cdna.
EST is a short sub-sequence of a cDNA sequence.
Used to identify gene transcripts, and are instrumental in gene discovery and in gene-sequence determination.
Approximately 74.2 million ESTs are available in public databases.
EST results from one-short sequencing of a cloned cDNA.
Low-quality fragments.
Length is approximately 500 to 800 nucleotides.
In a detail description of the two major blotting techniques. Right from its history to the result interpretation put forth in a concise way. Helps understand these protocols with ease.
Molecular techniques are major tools for the analysis of microorganisms.
Molecular methods varies with respect to discriminatory power, reproducibility, ease of use, and ease of interpretation.
Molecular marker technology in studies on plant genetic diversityChanakya P
A molecular marker is a molecule contained within a sample taken from an organism (biological markers) or other matter. It can be used to reveal certain characteristics about the respective source. DNA, for example, is a molecular marker containing information about genetic disorders, genealogy and the evolutionary history of life. Specific regions of the DNA (genetic markers) are used to diagnose the autosomal recessive genetic disorder cystic fibrosis, taxonomic affinity (phylogenetics) and identity (DNA Barcoding). Further, life forms are known to shed unique chemicals, including DNA, into the environment as evidence of their presence in a particular location.Other biological markers, like proteins, are used in diagnostic tests for complex neurodegenerative disorders, such as Alzheimer's disease. Non-biological molecular markers are also used, for example, in environmental studies.
The interface in a complex involves two structurally matched protein subunits, and the binding sites can be predicted by identifying structural matches at protein surfaces.
Identification of Protein–Ligand Binding Sites by Sequence & Identifying protein–ligand binding sites is an important process in drug discovery and structure-based drug design. Detecting protein–ligand binding sites is expensive and time-consuming by traditional experimental methods. Hence, computational approaches provide many effective strategies to deal with this issue. Recently, lots of computational methods are based on structure information on proteins. However, these methods are limited in the common scenario, where both the sequence of protein target is known and sufficient 3D structure information is available. Studies indicate that sequence-based computational approaches for predicting protein–ligand binding sites are more practical. Different methods were used to determine protein binding sites fir instance, chromatin immuno preciptitation assay ( ChIP),
Electrophoretic mobility shift assay (EMSA), Dnase footprinting assay etc.
Potential therapy derived from banana protein is effective against SARS-CoV-2Creative BioMart
On January 13, 2020, a paper was published online touting the creation of a possible therapy that could be used against all known strains of the flu. A week later, the first laboratory-confirmed case of SARS-CoV-2 sparked the two-and-a-half-year COVID-19 pandemic in the United States. Interestingly, before the virus temporarily halted their work, the international team of researchers for this flu paper also investigated treatments for the coronavirus.
CAR-T Cell Anti-Cancer Therapy Ushered in A Revolutionary Breakthrough!Creative BioMart
Chimeric antigen receptor T cells (CAR-T) and T cell antigen receptor chimeric T cells (TCR-T) are currently the "top stream" in adoptive T cell tumor immunotherapy. In particular, CAR-T therapy, which has been approved by the FDA, is rewriting the treatment paradigm of some hematological tumors.
In recent years, the increase in the number of multi-drug resistant pathogens and food safety have become serious global problems, and it is increasingly important to find or develop a new generation of antibacterial drugs or preservatives. Scientists have discovered that bacteria-produced bacteriocins can control clinically relevant susceptible and resistant bacteria, and purified bacteriocins can be added to foods as natural preservatives. Bacteriocins can be added to animal feeds as anti-pathogen additives to protect livestock from pathogen damage. In medicine, bacteriocin has the potential to replace antibiotics as antibacterial drugs and is a new type of anticancer drug.
Creative BioMart to Attend BIO International Convention June 13-16, 2022Creative BioMart
Creative BioMart, a leading biotechnology company supplying comprehensive protein products and services for the science community, recently announced that it will be attending the BIO International Convention in San Diego, California, from June 13-16, 2022.
Third Natural Eukaryotic Epigenetic Mark Found.pdfCreative BioMart
A group at the Marine Biology Laboratory (MBL) now found a third epigenetic mark in this freshwater invertebrate, Adineta vaga, which has previously been found only in bacteria. For the first time, a horizontally transferred gene has been shown to remodel gene regulatory systems in eukaryotes.
ELife: T Interferon Suppresses Inflammatory Diseases by Balancing the MicrobiomeCreative BioMart
The authors report that STAT1KO mice spontaneously develop inflammatory diseases characterized by bone marrow hyperplasia and splenic accumulation of hematopoietic stem cells.
Chemokines are small proteins, usually ~70–80 amino acid residues, with conserved sequence and structural features and expressed in tissues during normal immune surveillance or in response to injury or infection.
Creative BioMart has more than 20 years of experience in protein characterization. We have extensive experience in developing and establishing protein identification methods to help our customers generate the data they need to obtain the level of product identification required for regulatory submission. These characterization services are also part of our stability and product launch support services, where we provide critical identity, purity, and performance measurements.
The latest research has identified FGL1 as an immune inhibitory ligand of LAG-3, an immune inhibitory receptor that mediates T-cell suppression. Blocking FGL1/LAG-3 interaction improves anti-tumor immunity.
Creative BioMart has currently developed FGL1 proteins from multiple species. Their binding activity with LAG-3 protein has been verified by functional ELISA.
https://www.creativebiomart.net/resource/pdfdownload/dbd8e3ddfc158995b7e56f7277140c50.pdf
Protein quantification is divided into "total quantification method" of whole protein and "individual quantification method" of specific protein according to its purpose. It is an indispensable part of biological experiments.
To master the basic operation process of primary culture and subculture of mammalian cells, lay a foundation for the application of bioengineering in medicine.
Electrophoresis technology is the basic technology of molecular biology. This experiment helps better understanding of DNA electrophoresis tank and gel electrophoresis technology.
These simplified slides by Dr. Sidra Arshad present an overview of the non-respiratory functions of the respiratory tract.
Learning objectives:
1. Enlist the non-respiratory functions of the respiratory tract
2. Briefly explain how these functions are carried out
3. Discuss the significance of dead space
4. Differentiate between minute ventilation and alveolar ventilation
5. Describe the cough and sneeze reflexes
Study Resources:
1. Chapter 39, Guyton and Hall Textbook of Medical Physiology, 14th edition
2. Chapter 34, Ganong’s Review of Medical Physiology, 26th edition
3. Chapter 17, Human Physiology by Lauralee Sherwood, 9th edition
4. Non-respiratory functions of the lungs https://academic.oup.com/bjaed/article/13/3/98/278874
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!
- 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
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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
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
ARTIFICIAL INTELLIGENCE IN HEALTHCARE.pdfAnujkumaranit
Artificial intelligence (AI) refers to the simulation of human intelligence processes by machines, especially computer systems. It encompasses tasks such as learning, reasoning, problem-solving, perception, and language understanding. AI technologies are revolutionizing various fields, from healthcare to finance, by enabling machines to perform tasks that typically require human intelligence.
2. • The Electrophoretic Mobility Shift Assay (EMSA), or
gel shift assay is a basic and rapid method to detect
protein complex with nucleic acids.
3. • Originally utilized broadly in the investigation of
sequence-specific DNA-binding proteins such as
transcription factors, EMSA has been additionally
developed to explore DNA-protein interactions, RNA-
protein interactions.
4. • It is likewise connected to qualify an quantify
proteins that particularly tie to given nucleotides,
empowering to accommodate a wide range of
binding conditions.
5. Principles
• The purified protein and the cell crude
extract are usually incubated with the 32P
isotope-labeled DNA or RNA probe, and
the complex and the unbound probe are
isolated on the non-denaturing
polypropylene gel electrophoresis.
6. • DNA-complexes or RNA-complexes move slower than
non-bound probes. Isotope-labeled probes differ
depending on the binding protein studied, double-
stranded or single-stranded can both be ok. When
detecting DNA binding proteins such as
transcriptional regulatory factors, purified proteins,
partially purified proteins, or nuclear cell extracts can
be used.
7. • In the detection of RNA binding protein, according to
the location of the target RNA binding protein,
purified or partially purified protein, as well as
nuclear or cytoplasmic cell extract can also be used .
8. • The DNA or RNA fragments and oligonucleotide
fragments (specific) containing protein binding
sequences, and other non-relevant fragments (non-
specific), were used in competitive experiments to
determine the specificity of DNA or RNA-binding
proteins. Specific binding is determined according to
the characteristics and intensity of the complex in
the presence of competing specific and non-specific
fragments.
