This document discusses the use of PCR and restriction fragment length polymorphism (PRA) techniques to identify different species of non-tuberculous mycobacteria (NTM). NTM can cause various infections, especially in immunocompromised patients. The study aimed to evaluate the PRA method for identifying rapidly growing NTM species found in water and laboratory environments. Samples that tested positive for NTM were cultured and classified based on growth rate. Rapid growers were identified using PRA of the hsp65 gene. Results showed that PRA provided good discrimination of various NTM species and correlated with biochemical identification methods. The study concluded that PRA is a useful technique for rapid and easy differentiation of NTM species, which is
mRNA vaccine is a novel vaccine technology, which delivers mRNA that encoding the antigen protein of pathogen to the cell, and expresses the antigen protein, and then stimulates the immune response of the body.
Creative Biolabs has developed non-replicating mRNA vaccine platform, mRNA vaccine platform, mRNA pharmacology optimization platform, and and Self-amplifying mRNA vaccine platform to spport your vaccine researches. If you need more information about mRNA vaccine, please follow us.
CANFAST™ is widely used for both Primary Cell and established Cell Lines with minimal Cytotoxicity and optimal Transfection Efficiency. It is suitable for transfecting Human Cells as 293-HEK, BOSC 23 and HepG2, Mouse Cells as NIH3T3, P815, B16F10, Rat Cells as RBL2H3, PC12, and Hamster Cells as CHO-K1.
mRNA vaccine is a novel vaccine technology, which delivers mRNA that encoding the antigen protein of pathogen to the cell, and expresses the antigen protein, and then stimulates the immune response of the body.
Creative Biolabs has developed non-replicating mRNA vaccine platform, mRNA vaccine platform, mRNA pharmacology optimization platform, and and Self-amplifying mRNA vaccine platform to spport your vaccine researches. If you need more information about mRNA vaccine, please follow us.
CANFAST™ is widely used for both Primary Cell and established Cell Lines with minimal Cytotoxicity and optimal Transfection Efficiency. It is suitable for transfecting Human Cells as 293-HEK, BOSC 23 and HepG2, Mouse Cells as NIH3T3, P815, B16F10, Rat Cells as RBL2H3, PC12, and Hamster Cells as CHO-K1.
This slide is about the basics of mRNA-based therapy. The content includes: definition of mRNA, timeline of mRNA therapeutics, action mechanism and development strategies of mRNA drugs, therapeutic mRNA applications, and the related services provided by Creative Biolabs.
mechanism of resistance of antibiotics, ESBL, b lactums, enterobactericae, metallobactums, carbapenemases, types of mechanism of resistance, history of antibiotics and resistance
This slide is about the basics of mRNA-based therapy. The content includes: definition of mRNA, timeline of mRNA therapeutics, action mechanism and development strategies of mRNA drugs, therapeutic mRNA applications, and the related services provided by Creative Biolabs.
mechanism of resistance of antibiotics, ESBL, b lactums, enterobactericae, metallobactums, carbapenemases, types of mechanism of resistance, history of antibiotics and resistance
SLIDE CONTAIN BREIF NOTE ON PCR. IT CONTAINS 21 SLIDES INCLUDING, WHAT IS PCR? COMPONENTS, WORKING MECHANISM, APPLICATIONS, CONCLUSION, AND SOME REFRENCES, HISTORY ALSO
If a microbiologist is studying bacteria that premeditate, or break down, toxic wastes and wants to know which specific genes are active when that bacterium is degrading, say, PCBs, he would likely use a tool called the DNA microarray.
Microarrays enable scientists to monitor the activities of hundreds or thousands of genes at once. All microarrays (also called DNA chips or gene chips) work on the basic principle that complementary nucleotide sequences in DNA (and RNA) match up like the two halves of a piece of Velcro coming together.
Pattern of gene activity on a microarray chip.
A microarray consists of an orderly arrangement of bits of genetic material in super-tiny spots laid down in a grid on a suitable surface, often a glass slide with a specially chemically treated surface.
Food borne pathogens causes various diseases. So it is very important to detect them. Rapid methods help to detect pathogens in a very short period of time.
GENE CLONING,ITS HISTORY, NEW ADVENT IN GENE CLONING, PCR IMPORTANCE ,APPLICATION OF GENE CLONING,STEPS OF GENE CLONING,Antisense technology,Gene cloning in agriculture,Somatic cell therapy,Role of gene cloning in identification of genes responsible for human diseases,Synthesis of other recombinant human proteins and recombinant vaccines
Gene cloning in medicine,Recombinant protein from yeast,Problems with the production of recombinant protein in E.coli ,Expression of foreign genes in E.coli,Production of recombinant protein ,PCR can also be used to purify a gene,Obtaining a pure sample of a gene by cloning,Why gene cloning and PCR are so important,The advent of gene cloning and the polymerase
chain reaction.
Richard's entangled aventures in wonderlandRichard Gill
Since the loophole-free Bell experiments of 2020 and the Nobel prizes in physics of 2022, critics of Bell's work have retreated to the fortress of super-determinism. Now, super-determinism is a derogatory word - it just means "determinism". Palmer, Hance and Hossenfelder argue that quantum mechanics and determinism are not incompatible, using a sophisticated mathematical construction based on a subtle thinning of allowed states and measurements in quantum mechanics, such that what is left appears to make Bell's argument fail, without altering the empirical predictions of quantum mechanics. I think however that it is a smoke screen, and the slogan "lost in math" comes to my mind. I will discuss some other recent disproofs of Bell's theorem using the language of causality based on causal graphs. Causal thinking is also central to law and justice. I will mention surprising connections to my work on serial killer nurse cases, in particular the Dutch case of Lucia de Berk and the current UK case of Lucy Letby.
Cancer cell metabolism: special Reference to Lactate PathwayAADYARAJPANDEY1
Normal Cell Metabolism:
Cellular respiration describes the series of steps that cells use to break down sugar and other chemicals to get the energy we need to function.
Energy is stored in the bonds of glucose and when glucose is broken down, much of that energy is released.
Cell utilize energy in the form of ATP.
The first step of respiration is called glycolysis. In a series of steps, glycolysis breaks glucose into two smaller molecules - a chemical called pyruvate. A small amount of ATP is formed during this process.
Most healthy cells continue the breakdown in a second process, called the Kreb's cycle. The Kreb's cycle allows cells to “burn” the pyruvates made in glycolysis to get more ATP.
The last step in the breakdown of glucose is called oxidative phosphorylation (Ox-Phos).
It takes place in specialized cell structures called mitochondria. This process produces a large amount of ATP. Importantly, cells need oxygen to complete oxidative phosphorylation.
If a cell completes only glycolysis, only 2 molecules of ATP are made per glucose. However, if the cell completes the entire respiration process (glycolysis - Kreb's - oxidative phosphorylation), about 36 molecules of ATP are created, giving it much more energy to use.
IN CANCER CELL:
Unlike healthy cells that "burn" the entire molecule of sugar to capture a large amount of energy as ATP, cancer cells are wasteful.
Cancer cells only partially break down sugar molecules. They overuse the first step of respiration, glycolysis. They frequently do not complete the second step, oxidative phosphorylation.
This results in only 2 molecules of ATP per each glucose molecule instead of the 36 or so ATPs healthy cells gain. As a result, cancer cells need to use a lot more sugar molecules to get enough energy to survive.
Unlike healthy cells that "burn" the entire molecule of sugar to capture a large amount of energy as ATP, cancer cells are wasteful.
Cancer cells only partially break down sugar molecules. They overuse the first step of respiration, glycolysis. They frequently do not complete the second step, oxidative phosphorylation.
This results in only 2 molecules of ATP per each glucose molecule instead of the 36 or so ATPs healthy cells gain. As a result, cancer cells need to use a lot more sugar molecules to get enough energy to survive.
introduction to WARBERG PHENOMENA:
WARBURG EFFECT Usually, cancer cells are highly glycolytic (glucose addiction) and take up more glucose than do normal cells from outside.
Otto Heinrich Warburg (; 8 October 1883 – 1 August 1970) In 1931 was awarded the Nobel Prize in Physiology for his "discovery of the nature and mode of action of the respiratory enzyme.
WARNBURG EFFECT : cancer cells under aerobic (well-oxygenated) conditions to metabolize glucose to lactate (aerobic glycolysis) is known as the Warburg effect. Warburg made the observation that tumor slices consume glucose and secrete lactate at a higher rate than normal tissues.
Slide 1: Title Slide
Extrachromosomal Inheritance
Slide 2: Introduction to Extrachromosomal Inheritance
Definition: Extrachromosomal inheritance refers to the transmission of genetic material that is not found within the nucleus.
Key Components: Involves genes located in mitochondria, chloroplasts, and plasmids.
Slide 3: Mitochondrial Inheritance
Mitochondria: Organelles responsible for energy production.
Mitochondrial DNA (mtDNA): Circular DNA molecule found in mitochondria.
Inheritance Pattern: Maternally inherited, meaning it is passed from mothers to all their offspring.
Diseases: Examples include Leber’s hereditary optic neuropathy (LHON) and mitochondrial myopathy.
Slide 4: Chloroplast Inheritance
Chloroplasts: Organelles responsible for photosynthesis in plants.
Chloroplast DNA (cpDNA): Circular DNA molecule found in chloroplasts.
Inheritance Pattern: Often maternally inherited in most plants, but can vary in some species.
Examples: Variegation in plants, where leaf color patterns are determined by chloroplast DNA.
Slide 5: Plasmid Inheritance
Plasmids: Small, circular DNA molecules found in bacteria and some eukaryotes.
Features: Can carry antibiotic resistance genes and can be transferred between cells through processes like conjugation.
Significance: Important in biotechnology for gene cloning and genetic engineering.
Slide 6: Mechanisms of Extrachromosomal Inheritance
Non-Mendelian Patterns: Do not follow Mendel’s laws of inheritance.
Cytoplasmic Segregation: During cell division, organelles like mitochondria and chloroplasts are randomly distributed to daughter cells.
Heteroplasmy: Presence of more than one type of organellar genome within a cell, leading to variation in expression.
Slide 7: Examples of Extrachromosomal Inheritance
Four O’clock Plant (Mirabilis jalapa): Shows variegated leaves due to different cpDNA in leaf cells.
Petite Mutants in Yeast: Result from mutations in mitochondrial DNA affecting respiration.
Slide 8: Importance of Extrachromosomal Inheritance
Evolution: Provides insight into the evolution of eukaryotic cells.
Medicine: Understanding mitochondrial inheritance helps in diagnosing and treating mitochondrial diseases.
Agriculture: Chloroplast inheritance can be used in plant breeding and genetic modification.
Slide 9: Recent Research and Advances
Gene Editing: Techniques like CRISPR-Cas9 are being used to edit mitochondrial and chloroplast DNA.
Therapies: Development of mitochondrial replacement therapy (MRT) for preventing mitochondrial diseases.
Slide 10: Conclusion
Summary: Extrachromosomal inheritance involves the transmission of genetic material outside the nucleus and plays a crucial role in genetics, medicine, and biotechnology.
Future Directions: Continued research and technological advancements hold promise for new treatments and applications.
Slide 11: Questions and Discussion
Invite Audience: Open the floor for any questions or further discussion on the topic.
(May 29th, 2024) Advancements in Intravital Microscopy- Insights for Preclini...Scintica Instrumentation
Intravital microscopy (IVM) is a powerful tool utilized to study cellular behavior over time and space in vivo. Much of our understanding of cell biology has been accomplished using various in vitro and ex vivo methods; however, these studies do not necessarily reflect the natural dynamics of biological processes. Unlike traditional cell culture or fixed tissue imaging, IVM allows for the ultra-fast high-resolution imaging of cellular processes over time and space and were studied in its natural environment. Real-time visualization of biological processes in the context of an intact organism helps maintain physiological relevance and provide insights into the progression of disease, response to treatments or developmental processes.
In this webinar we give an overview of advanced applications of the IVM system in preclinical research. IVIM technology is a provider of all-in-one intravital microscopy systems and solutions optimized for in vivo imaging of live animal models at sub-micron resolution. The system’s unique features and user-friendly software enables researchers to probe fast dynamic biological processes such as immune cell tracking, cell-cell interaction as well as vascularization and tumor metastasis with exceptional detail. This webinar will also give an overview of IVM being utilized in drug development, offering a view into the intricate interaction between drugs/nanoparticles and tissues in vivo and allows for the evaluation of therapeutic intervention in a variety of tissues and organs. This interdisciplinary collaboration continues to drive the advancements of novel therapeutic strategies.
Nutraceutical market, scope and growth: Herbal drug technologyLokesh Patil
As consumer awareness of health and wellness rises, the nutraceutical market—which includes goods like functional meals, drinks, and dietary supplements that provide health advantages beyond basic nutrition—is growing significantly. As healthcare expenses rise, the population ages, and people want natural and preventative health solutions more and more, this industry is increasing quickly. Further driving market expansion are product formulation innovations and the use of cutting-edge technology for customized nutrition. With its worldwide reach, the nutraceutical industry is expected to keep growing and provide significant chances for research and investment in a number of categories, including vitamins, minerals, probiotics, and herbal supplements.
The increased availability of biomedical data, particularly in the public domain, offers the opportunity to better understand human health and to develop effective therapeutics for a wide range of unmet medical needs. However, data scientists remain stymied by the fact that data remain hard to find and to productively reuse because data and their metadata i) are wholly inaccessible, ii) are in non-standard or incompatible representations, iii) do not conform to community standards, and iv) have unclear or highly restricted terms and conditions that preclude legitimate reuse. These limitations require a rethink on data can be made machine and AI-ready - the key motivation behind the FAIR Guiding Principles. Concurrently, while recent efforts have explored the use of deep learning to fuse disparate data into predictive models for a wide range of biomedical applications, these models often fail even when the correct answer is already known, and fail to explain individual predictions in terms that data scientists can appreciate. These limitations suggest that new methods to produce practical artificial intelligence are still needed.
In this talk, I will discuss our work in (1) building an integrative knowledge infrastructure to prepare FAIR and "AI-ready" data and services along with (2) neurosymbolic AI methods to improve the quality of predictions and to generate plausible explanations. Attention is given to standards, platforms, and methods to wrangle knowledge into simple, but effective semantic and latent representations, and to make these available into standards-compliant and discoverable interfaces that can be used in model building, validation, and explanation. Our work, and those of others in the field, creates a baseline for building trustworthy and easy to deploy AI models in biomedicine.
Bio
Dr. Michel Dumontier is the Distinguished Professor of Data Science at Maastricht University, founder and executive director of the Institute of Data Science, and co-founder of the FAIR (Findable, Accessible, Interoperable and Reusable) data principles. His research explores socio-technological approaches for responsible discovery science, which includes collaborative multi-modal knowledge graphs, privacy-preserving distributed data mining, and AI methods for drug discovery and personalized medicine. His work is supported through the Dutch National Research Agenda, the Netherlands Organisation for Scientific Research, Horizon Europe, the European Open Science Cloud, the US National Institutes of Health, and a Marie-Curie Innovative Training Network. He is the editor-in-chief for the journal Data Science and is internationally recognized for his contributions in bioinformatics, biomedical informatics, and semantic technologies including ontologies and linked data.
Multi-source connectivity as the driver of solar wind variability in the heli...Sérgio Sacani
The ambient solar wind that flls the heliosphere originates from multiple
sources in the solar corona and is highly structured. It is often described
as high-speed, relatively homogeneous, plasma streams from coronal
holes and slow-speed, highly variable, streams whose source regions are
under debate. A key goal of ESA/NASA’s Solar Orbiter mission is to identify
solar wind sources and understand what drives the complexity seen in the
heliosphere. By combining magnetic feld modelling and spectroscopic
techniques with high-resolution observations and measurements, we show
that the solar wind variability detected in situ by Solar Orbiter in March
2022 is driven by spatio-temporal changes in the magnetic connectivity to
multiple sources in the solar atmosphere. The magnetic feld footpoints
connected to the spacecraft moved from the boundaries of a coronal hole
to one active region (12961) and then across to another region (12957). This
is refected in the in situ measurements, which show the transition from fast
to highly Alfvénic then to slow solar wind that is disrupted by the arrival of
a coronal mass ejection. Our results describe solar wind variability at 0.5 au
but are applicable to near-Earth observatories.
2. Introduction
Hsp 65
• have been
considered as
useful
phylogenetic
markers in several
eubacterial
genera
• Highly conserved
primary structures
mycobacterium
• A genus of gram
positive and
aerobic bacilli.
• Tuberculous:
group of 4 species
(koch bacilli)
• NTM: mainly
affect
inmmunocompro
mised patients
PAGE
• It is a widely used
technique to
separate proteins
according to their
electrophoretic
mobility
• Particles are
separate by their
weight and charge
PRA
• The method
involves
restriction
enzyme analysis
of PCR products
obtained with
primers common
to all
mycobacteria.
3. Introduction
NTM can
cause:
• Pulmonary infection, lymph node infection, brain,
bone, kydney, genital tract and skin infection
Identification of
Hsp65
Use of
techniques
like PRA
• contains epitopes that are unique as well as
epitopes that are common to various species
of mycobacterium
• Will be very useful to identify
different species of NTM in a faster
and easly way.
4. OBJECTIVE
• The aim of the present study was to
evaluate the PRA method and to extend
it to other mycobacterial species,
especially rapidly growing
nonchromogenic mycobacteria, often
found in tap water and laboratory
environment.
5. MATERIALES Y MÉTODOS
Estudio realizado entre
octubre 2013 y
diciembre de 2014
recolección de muestras
positivas para NTM
Bacterias puestas en
medio de cultivo y
analizadas
Según la velocidad de
crecimiento se
clasificaron en:
NTM de crecimiento
rápido y de crecimiento
lento
NTM de crecimiento
rápido fueron
identificadas por PARA
(genes que codifican
Hsp 65)
6. Extracción de
DNA Fundamento: el reactivo se une cationes como
Mg2 +, que es un cofactor esencial para las
ADNasas. Por lo que protege la muestra de las
ADNasas que podrían permanecer activas después
de la ebullición y podría posteriormente degradar el
ADN
REACTIVO CHELEX
Uso: protege el DNA, evitando su degradación
Se añade el reactivo, se pone en vortex y se
centrifuga
7. PCR
(reacción en cadena de
la polimeraza)
FUNDAMENTO
Es una técnica utilizada para la amplificación
de un gen o de un fragmento de DNA,
permite la obtención de muchas copias de
una secuencia de DNA mediante la
replicación de la hebra sencilla de DNA por
una DNA polimeraza a partir de un primer
USO: se amplificaron las secuencias que
codifican para la Hsp 65
TB11: 50-ACCAACGATGGTGTGTCCAT-30
8. ANÁLISIS DE
RESTRICCIÓN
FUNDAMENTO:
Las enzimas de restricción rompen la cadena de
DNA al reconocer una secuencia específica de bases
y consiste en deshacer la unión que forma el grupo
fosfato entre dos moléculas de desoxirribosa de la
hebra de DNA
USO: las enzimas de restricción fueron
utilizadas para cortar segmentos específicos de
la cadena
13. AUTHOR ARTICLE YES, NO
Steingrube et al Bst II, Hae III along with other enzymes AciI and CfoI
gave the best separation of rapidly growing
mycobacteria.12
Ringuet et al The results of amplification of hsp65 gene followed by
restriction enzyme analysis of rapid grower
mycobacterium helped in discriminating 8 different
types of rapid grower mycobacterium which were well
correlated with biochemical identification results
Buchanan et al. In this study, the highly conserved hsp-65 gene was
targeted for amplifications. It has a high discriminating
power to differentiate species of Mycobacteria
Telenti et al The restriction enzyme digestion
patterns obtained in the present study provides
species
separation similar to those described by Telenti et al.
DISCUSSION
14. CONCLUSIONS
• PRA is a very good technique to determinate
and differenciate different species of NTM in a
easy and fast way.
• With the increase of immunosuppressed people
for various reasons, the differentiation of the
NTM species is indispensable to be able to treat
in a timely manner the infections caused by
these bacteria and prevent the alterations that
they generate.
Editor's Notes
With the help of 26 different
band patterns, the rapid growers were divided into 8 species
(Table 3)
4. A pesar de que el autor si describe que las otras dos enzimas de restricción pueden generar mayor especificidad, las únicas enzimas de r usadas en este art fueron y según steingrube tienen entre un 94 y un 100% de especificidad dependiendo del tipo de micobacteria y la aci I y la Cfol solo son especificas cuando se usan las ptras dos, sino la separación es menos optima
