Circular dichroism spectroscopy measures the differential absorption of left and right circularly polarized light by chiral molecules. When light passes through an optically active substance, the left and right circular polarizations are absorbed to different extents. A CD spectrometer contains a light source, monochromator, polarizer, photoelastic modulator and detector. It measures the CD signal as a function of wavelength, providing information about secondary structure of proteins and nucleic acids. CD spectroscopy requires minimal sample amounts and can quickly analyze secondary structure without crystallization. It is useful for studying protein folding, ligand binding and environmental effects on structure.
Circular Dichroism Spectroscopy
Introduction
Amino Acid Structure & Polarity
Protein Structures
Polarization of Light
Circular Dichroism
Measurement of Circular Dichroism
Instrumentation For CD Spectropolarimeter
CD Spectrum
CD Spectra of Protein Secondary Structures
Other - CD Based Techniques
Conclusion
References
described about optical activity, specific rotation, angle of rotation and circular dichroism and differences between ORD and CD and applications of ORD AND CD,
A detail and straight forward information about th CD and ORD
and Also about the polarization of light i.e. plane polarized light and circular polarized light
Circular Dichroism Spectroscopy
Introduction
Amino Acid Structure & Polarity
Protein Structures
Polarization of Light
Circular Dichroism
Measurement of Circular Dichroism
Instrumentation For CD Spectropolarimeter
CD Spectrum
CD Spectra of Protein Secondary Structures
Other - CD Based Techniques
Conclusion
References
described about optical activity, specific rotation, angle of rotation and circular dichroism and differences between ORD and CD and applications of ORD AND CD,
A detail and straight forward information about th CD and ORD
and Also about the polarization of light i.e. plane polarized light and circular polarized light
Transmission electron microscopy (TEM)- by sivasangari Shanmugam. Transmission electron microscopy (TEM) is a technique used to observe the features of very small specimens.
Isoelectric focusing electrophoresis- Principle , procedure and applicationsJaskiranKaur72
IEF separates amphoteric compounds, such as proteins, with increased resolution in a medium possessing a stable pH gradient. The protein becomes “focused” at a point on the gel as it migrates to a zone where the pH of the gel matches the protein's pI. At this point, the charge of the protein becomes zero and its migration ceases.
It is a subtype of the gel electrophoresis whereby the normal gel is replaced with polyacrylamide gels used as support media.
Gels are made by free radical-induced polymerization of acrylamide and N,N’-Methylenebisacrylamide.
It is the most widely used technique of electrophoresis.
Sedimentation for determining molecular weight of macromoleculesShubhangiSuri1
Process of sedimentation with mechanism of action and mathematical derivations, different methods for separation of macromolecules by sedimentation, viscometry vs sedimentation
Electron Spray Ionization (ESI) and its ApplicationsNisar Ali
In this slide ,You will get to learn Electron Spray Ionization (ESI) technique used in Mass Spectroscopy and its Various Application in Pharmaceutical Drug Analysis.
SEM is a type of electron microscope designed for directly studying the surfaces of solid objects, that utilizes a beam of focused electron of relatively low energy as an electron probe that is scanned in a regular manner over the specimen.
Transmission electron microscopy (TEM)- by sivasangari Shanmugam. Transmission electron microscopy (TEM) is a technique used to observe the features of very small specimens.
Isoelectric focusing electrophoresis- Principle , procedure and applicationsJaskiranKaur72
IEF separates amphoteric compounds, such as proteins, with increased resolution in a medium possessing a stable pH gradient. The protein becomes “focused” at a point on the gel as it migrates to a zone where the pH of the gel matches the protein's pI. At this point, the charge of the protein becomes zero and its migration ceases.
It is a subtype of the gel electrophoresis whereby the normal gel is replaced with polyacrylamide gels used as support media.
Gels are made by free radical-induced polymerization of acrylamide and N,N’-Methylenebisacrylamide.
It is the most widely used technique of electrophoresis.
Sedimentation for determining molecular weight of macromoleculesShubhangiSuri1
Process of sedimentation with mechanism of action and mathematical derivations, different methods for separation of macromolecules by sedimentation, viscometry vs sedimentation
Electron Spray Ionization (ESI) and its ApplicationsNisar Ali
In this slide ,You will get to learn Electron Spray Ionization (ESI) technique used in Mass Spectroscopy and its Various Application in Pharmaceutical Drug Analysis.
SEM is a type of electron microscope designed for directly studying the surfaces of solid objects, that utilizes a beam of focused electron of relatively low energy as an electron probe that is scanned in a regular manner over the specimen.
Optical rotatory dispersion is the variation in the optical rotation of a substance with a change in the wavelength of light.
For wavelengths that are absorbed by the optically active sample, the two circularly polarized components will be absorbed to differing extents. This unequal absorption is known as circular dichroism.
Infrared spectroscopy (IR spectroscopy or vibrational spectroscopy) involves the interaction of infrared radiation with matter. It covers a range of techniques, mostly based on absorption spectroscopy. As with all spectroscopic techniques, it can be used to identify and study chemicals
Richard's aventures in two entangled wonderlandsRichard 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.
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.
THE IMPORTANCE OF MARTIAN ATMOSPHERE SAMPLE RETURN.Sérgio Sacani
The return of a sample of near-surface atmosphere from Mars would facilitate answers to several first-order science questions surrounding the formation and evolution of the planet. One of the important aspects of terrestrial planet formation in general is the role that primary atmospheres played in influencing the chemistry and structure of the planets and their antecedents. Studies of the martian atmosphere can be used to investigate the role of a primary atmosphere in its history. Atmosphere samples would also inform our understanding of the near-surface chemistry of the planet, and ultimately the prospects for life. High-precision isotopic analyses of constituent gases are needed to address these questions, requiring that the analyses are made on returned samples rather than in situ.
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.
(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.
Professional air quality monitoring systems provide immediate, on-site data for analysis, compliance, and decision-making.
Monitor common gases, weather parameters, particulates.
Seminar of U.V. Spectroscopy by SAMIR PANDASAMIR PANDA
Spectroscopy is a branch of science dealing the study of interaction of electromagnetic radiation with matter.
Ultraviolet-visible spectroscopy refers to absorption spectroscopy or reflect spectroscopy in the UV-VIS spectral region.
Ultraviolet-visible spectroscopy is an analytical method that can measure the amount of light received by the analyte.
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.
Earliest Galaxies in the JADES Origins Field: Luminosity Function and Cosmic ...Sérgio Sacani
We characterize the earliest galaxy population in the JADES Origins Field (JOF), the deepest
imaging field observed with JWST. We make use of the ancillary Hubble optical images (5 filters
spanning 0.4−0.9µm) and novel JWST images with 14 filters spanning 0.8−5µm, including 7 mediumband filters, and reaching total exposure times of up to 46 hours per filter. We combine all our data
at > 2.3µm to construct an ultradeep image, reaching as deep as ≈ 31.4 AB mag in the stack and
30.3-31.0 AB mag (5σ, r = 0.1” circular aperture) in individual filters. We measure photometric
redshifts and use robust selection criteria to identify a sample of eight galaxy candidates at redshifts
z = 11.5 − 15. These objects show compact half-light radii of R1/2 ∼ 50 − 200pc, stellar masses of
M⋆ ∼ 107−108M⊙, and star-formation rates of SFR ∼ 0.1−1 M⊙ yr−1
. Our search finds no candidates
at 15 < z < 20, placing upper limits at these redshifts. We develop a forward modeling approach to
infer the properties of the evolving luminosity function without binning in redshift or luminosity that
marginalizes over the photometric redshift uncertainty of our candidate galaxies and incorporates the
impact of non-detections. We find a z = 12 luminosity function in good agreement with prior results,
and that the luminosity function normalization and UV luminosity density decline by a factor of ∼ 2.5
from z = 12 to z = 14. We discuss the possible implications of our results in the context of theoretical
models for evolution of the dark matter halo mass function.
This pdf is about the Schizophrenia.
For more details visit on YouTube; @SELF-EXPLANATORY;
https://www.youtube.com/channel/UCAiarMZDNhe1A3Rnpr_WkzA/videos
Thanks...!
2. WHAT IS CIRCULAR DICHROISM?
Circular dichroism is observed when an optically active matter absorbs left and right
hand circular polarized slightly differently.
Linearly polarised light is light whose oscillations are confined to a single plane.
All polarised light states can be described as a sum of two linearly polarised states at
right angles to each other, usually referenced to the viewer as vertically and
horizontally polarised light.
Verticallypolarisedlight
2
4. If one of the polarised states is out of phase with the other by a quarter-wave, the
resultant will be a helix and is known as circularly polarised light (CPL). The helices
can be either right-handed (R-CPL) or left-handed (L-CPL).
Left circularly polarised
light
Right circularly polarised
light
4
5. Circular Dichroism (CD) is measured with a CD spectropolarimeter, which is
relatively expensive (~$70k).
The instrument needs to be able to measure accurately in the far UV at wavelengths
down to 190-170 nm. In addition, the difference in left and right handed absorbance
A(l)- A(r) is very small (usually in the range of 0.0001) , therefore should be sensitive .
The CD is a function of wavelength.
When circularly polarized light passes through an absorbing optically active medium,
the speeds between right and left polarizations differ (cL ≠ cR) as well as their
wavelength (λL ≠ λR) and the extent to which they are absorb (εL ≠ εR). Circular
dichroism is the difference εL ≠ εR .
CD spectra for distinct types of secondary structure present in peptides, proteins and
nucleic acids are different. The analysis of CD spectra can therefore yield valuable
information about secondary structure of biological macromolecules.
5
6. CD Instrumentation
So we needmost of the same things as for a UV/visible:
Lamp -Xe or Xe/Hg .
Monochromator.
Polarizer.
Photo-elastic modulator.
Sample compartment.
Detector (photomultiplier).
6
8. The Chirascan™ monochromator uses two synthetic,
single‐crystal quartz prisms instead of the diffraction gratings that most people are
familiar with from normal absorbance spectrophotometers.
Quartz prisms are more efficient than diffraction gratings for a very wide range of
wavelengths, particularly in the UV.
Quartz is also birefringent and the prisms not only disperse light into the
component wavelengths but also, because of their birefringence, disperse the linearly
polarised components, one of which is selected for conversion to circularly polarised
light.
A further advantage of prisms is they do not pass second‐order multiples of the
desired wavelength, which is a major source of stray‐light in grating‐based
monochromators.
8
9. Photo-elastic modulator:
This is a piezoelectric element cemented to a block of fused silica.
At rest, when the piezoelectric element is not oscillating, the silica block is not
birefringent . when driven, the piezoelectric element oscillates at its resonance
frequency (typically around 50 kHz), and induces stress in the silica in such a way
that it becomes birefringent.
The alternating stress turns the fused silica element into a dynamic quarter‐wave
plate, retarding first vertical with respect to horizontal components of the incident
linearly polarised light by a quarter‐wave and then vice versa, producing left‐ and
then right‐ circularly polarised light at the drive frequency.
The amplitude of the oscillation is tuned so that the retardation is appropriate for the
wavelength of light passing through the silica block.
9
11. Operation of CD spectrometer.
11
A CD active sample will absorb either L-CPL or R-CPL preferentially
Consequently there is only a steady output from the light detector.
With no CD active sample, the L-CPL and R-CPL have equal light intensities.
The photo elastic modulator converts the linearly polarised beam into left and right
circularly polarised light altering at 50 KHz.
The monochromator outputs linearly polarised light of a single
wavelength(monochromatic).
12. 12
Plot of CD signal Vs. wavelength of light gives the CD spectrum.
The vAC signal is divided by vDC signal. The siganl is multiplied by a calibration
factor (G) to give units of milidegrees or absorbance.
The average of the light intensity over time is measured and is termed as vDC. This
can be used to scale the changes in CD measured by vAC signal.
Difference in the intensity of L-CPL and R-CPL components is measured by amplifier
tuned to the PEM frequency.
The result of the unequal amount of L-CPL and R-CPL reaching the detector , is a
signal that varies with the frequency of the PEM (50 KHz).
13. Sample preparation
Additives, buffers and stabilizing compounds: Any compound which absorbs in
the region of interest (250 - 190 nm) should be avoided.
A buffer or detergent or other chemical should not be used unless it can be shown
that the compound in question will not mask the protein signal.
Protein solution: From the above follows that the protein solution should contain
only those chemicals necessary to maintain protein stability, and at the lowest
concentrations possible. Avoid any chemical that is unnecessary for protein
stability/solubility. The protein itself should be as pure as possible, any additional
protein or peptide will contribute to the CD signal.
13
14. Contaminants: Unfolded protein, peptides, particulate matter
(scattering particles), anything that adds significant noise (or artificial
signal contributions) to the CD spectrum must be avoided. Filtering of
the solutions (0.02 um syringe filters) may improve signal to noise ratio.
Nitrogen purging: the function of purging the CD instrument with
nitrogen is to remove oxygen from the lamp housing, monochromator,
and the sample chamber. The reason for removing oxygen is that
oxygen absorbs deep UV light, thus reducing the light available for the
measurement.
14
15. Typical Conditions for CD
• Protein Concentration: 0.25 mg/ml
• Cell Path Length: 1 mm
• Volume 400 μl
• Need very little sample 0.1 mg
• Concentration reasonable
• Stabilizers (Metal ions, etc.): minimum
• Buffer Concentration : 5 mM or as low as
possible while maintaining protein stability
• A structural biology method that can give real answers in a day
15
17. ADVANTAGES
Simple and quick experiment.
No extensive preparation..
Relatively low
concentrations/amounts of
sample.
Microsecond time resolution.
Any size of macromolecule.
DISADVANTAGES
Difficult to quantitate
similarity or differences.
Certain buffer components
absorb strongly in Far-UV &
can cause interference.
17
18. Applications
CD spectroscopy has wide range of applications in many different fields.
UV CD – to investigate the secondary structure of proteins that cannot be crystallized.
UV/ VIS CD – to investigate charge-transfer transitions.
Near IR CD – to investigate geometric and electronic structure by probing metal
transitions.
Investigation of the effect of drug binding on protein secondary structure.
Studies of the effects of environment on protein structure.
Study of ligand -induced conformational changes.
18
19. References
Applications of Circular Dichroism, International Journal of Molecular Sciences ISSN 1422-
0067.
www.mdpi.com/journal/ijms - visited on 22/11/2014.
Understanding Circular Dichroism , Applied Photo physics Ltd- www.photophysics.com -
22/11/2014.
Protein Secondary Structure and Circular Dichroism : A Practical Guide, W.C.Johnson,
PROTEINS 7, 205-214 (1990).
Circular Dichroism and Optical Rotary Dispersion of Proteins and Polypeptides, A.J.Alder,
N.J.Greenfield and G.D.Fasman, Meth. Enzymology 27, 675 (1973).
www.wikipedia.com – 24/11/2014.
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