Structure and Dynamics of Macromolecules Absorption

and Fluorescence Studies 1st Edition J.R. Albani

direct download

https://ebookfinal.com/download/structure-and-dynamics-of-
macromolecules-absorption-and-fluorescence-studies-1st-edition-j-r-
albani/

★★★★★
4.7 out of 5.0 (84 reviews )

PDF Instantly Ready

ebookfinal.com
 Structure and Dynamics of Macromolecules Absorption and
Fluorescence Studies 1st Edition J.R. Albani Pdf Download

EBOOK

Available Formats

■ PDF eBook Study Guide Ebook

EXCLUSIVE 2025 ACADEMIC EDITION – LIMITED RELEASE

Available Instantly Access Library

Here are some recommended products for you. Click the link to
download, or explore more at ebookfinal

Principles and Applications of Fluorescence Spectroscopy

1st Edition Jihad Rene Albani

https://ebookfinal.com/download/principles-and-applications-of-
fluorescence-spectroscopy-1st-edition-jihad-rene-albani/

The Quantitative Analysis of the Dynamics and Structure of

Terminologies 1st Edition Kyo Kageura

https://ebookfinal.com/download/the-quantitative-analysis-of-the-
dynamics-and-structure-of-terminologies-1st-edition-kyo-kageura/

Structure and Dynamics An Atomic View of Materials 1st

Edition Martin T. Dove

https://ebookfinal.com/download/structure-and-dynamics-an-atomic-view-
of-materials-1st-edition-martin-t-dove/

Dynamics of Structure and Foundation A Unified Approach 2

Applications 1st Edition Indrajit Chowdhury

https://ebookfinal.com/download/dynamics-of-structure-and-foundation-
a-unified-approach-2-applications-1st-edition-indrajit-chowdhury/
Structure and dynamics an atomic view of materials XD US
Martin T Dove

https://ebookfinal.com/download/structure-and-dynamics-an-atomic-view-
of-materials-xd-us-martin-t-dove/

Modification and Blending of Synthetic and Natural

Macromolecules 1st Edition Ciardelli F.

https://ebookfinal.com/download/modification-and-blending-of-
synthetic-and-natural-macromolecules-1st-edition-ciardelli-f/

Abstract algebra Smith J.R

https://ebookfinal.com/download/abstract-algebra-smith-j-r/

Physical Properties of Macromolecules 1st Edition Laurence

A. Belfiore

https://ebookfinal.com/download/physical-properties-of-
macromolecules-1st-edition-laurence-a-belfiore/

Economic Dynamics Theory Games and Empirical Studies 1st

Edition Chester W. Hurlington

https://ebookfinal.com/download/economic-dynamics-theory-games-and-
empirical-studies-1st-edition-chester-w-hurlington/
Structure and Dynamics of Macromolecules Absorption
and Fluorescence Studies 1st Edition J.R. Albani Digital
Instant Download
Author(s): J.R. Albani
ISBN(s): 9780444514493, 044451449X
Edition: 1
File Details: PDF, 32.02 MB
Year: 2005
Language: english
Cover illustration:
Energy transfer between Trp residues of apocytochrome b2 core and 2,p-toluidinyl-naphthalene-6-
sulfonate (TNS). Emission of Trp residues decreases in presence of increased amount of TNS. Source:
Albani, J.R. 1985, Arch. Biochem. Biophys. 243, 292-297.
 STRUCTURE AND DYNAMICS
OF MACROMOLECULES: ABSORPTION
AND FLUORESCENCE STUDIES
ELSEVIER B.V. ELSEVIER Inc. ELSEVIER Ltd. ELSEVIER Ltd.
Sara Burgerhartstraat 25 525 B Street The Boulevard 84 Theobalds Road
P.O. Box 211,1000 AE Suite 1900, San Diego Langford Lane, Kidlington, London WC1X8RR
Amsterdam, The Netherlands CA 92101-4495, USA Oxford OX5 1GB, UK UK

2004 Elsevier B.V. All rights reserved.

This work is protected under copyright by Elsevier B.V., and the following terms and conditions apply to its use:

Photocopying
Single photocopies of single chapters may be made for personal use as allowed by national copyright laws. Permission of the Publisher
and payment of a fee is required for all other photocopying, including multiple or systematic copying, copying for advertising or
promotional purposes, resale, and all forms of document delivery. Special rates are available for educational institutions that wish to
make photocopies for non-profit educational classroom use.

Permissions may be sought directly from Elsevier's Rights Department in Oxford, UK: phone (+44) 1865 843830, fax (+44)
1865 853333, e-mail: [email protected]. Requests may also be completed on-line via the Elsevier homepage (http://
www.elsevier.com/locate/permissions).

In the USA, users may clear permissions and make payments through the Copyright Clearance Center, Inc., 222 Rosewood Drive,
Danvers, MA 01923, USA; phone: (+1) (978) 7508400, fax: (+1) (978) 7504744, and in the UK through the Copyright Licensing
Agency Rapid Clearance Service (CLARCS), 90 Tottenham Court Road, London W1P OLP, UK; phone: (+44) 20 7631 5555; fax:
(+44) 20 7631 5500. Other countries may have a local reprographic rights agency for payments.

Derivative Works
Tables of contents may be reproduced for internal circulation, but permission of the Publisher is required for external resale or
distribution of such material. Permission of the Publisher is required for all other derivative works, including compilations and
translations.

Electronic Storage or Usage

Permission of the Publisher is required to store or use electronically any material contained in this work, including any chapter or
part of a chapter.

Except as outlined above, no part of this work may be reproduced, stored in a retrieval system or transmitted in any form or by any
means, electronic, mechanical, photocopying, recording or otherwise, without prior written permission of the Publisher.
Address permissions requests to: Elsevier's Rights Department, at the fax and e-mail addresses noted above.

Notice
No responsibility is assumed by the Publisher for any injury and/or damage to persons or property as a matter of products liability,
negligence or otherwise, or from any use or operation of any methods, products, instructions or ideas contained in the material
herein. Because of rapid advances in the medical sciences, in particular, independent verification of diagnoses and drug dosages
should be made.

First edition 2004

Library of Congress Cataloging in Publication Data

A catalog record is available from the Library of Congress.

British Library Cataloguing in Publication Data

A catalogue record is available from the British Library.

ISBN: 0-444-51449-X

@ The paper used in this publication meets the requirements of ANSI/NISO Z39.48-1992 (Permanence of Paper).
Printed in The Netherlands.
 V

Preface

Absorption and fluorescence spectroscopies are widely used to characterize the structure
and dynamics of biological molecules. It is now clearly established that biological function of
a molecule depends on its structure and / or dynamics.
This book deals with the fundamentals of the absorption and fluorescence along with
applications on the characterization of the structure and dynamics of macromolecules. The
example chosen from the literature are discussed, and in many cases we show that it is
possible to interpret the same result in two different and contradicted ways.
The examples given in the chapter dealing with the origin of proteins fluorescence clearly
rule out the existence of a general model describing this origin. Also, in one of the chapters,
we describe fluorescence experiments that led us to obtain a description of the tertiary
structure of (Xi- acid glycoprotein. In the same chapter, we added the first results we obtained
recently with X-ray and electron microscopy on the protein and its carbohydrate residues.
Finally, the last chapter describes novel experiments showing that animal and vegetal species
and varieties within the same species have characteristic fluorescence fingerprints.
This book will be useful to students, professors and researchers who want to discover, to
learn, to teach or to use absorption and fluorescence spectroscopies.
We were not able to cite the works of all the colleagues using absorption and fluorescence
spectroscopies in their research. However, we believe that every work carried out in the field
of these two techniques is important to find out their performance and their limits.
Finally, the author wishes to thank the designer Mr. Roger Boudiab who redraws most of
the graphics already published in the literature and which is used in this book.
This page is intentionally left blank
 vii

Contents

Preface v

1. Light absorption by a molecule

1. Jablonski diagram or diagram of electronic transitions 1
2. Singlet and triplet states 2
3. Forbidden and non forbidden transitions 7
4. Reading the Jablonski diagram 7
5. Chemical bonds 8
5a. Atomic and molecular orbitals 8
5b. The coordinated bond 9
6. Absorption spectroscopy 11
6a. Origin and properties of the absorption spectra 11
6b. Beer-Lambert-Bouguer law 14
6c. Determination of the molar extinction coefficient of proteins 18
6d. Effect of high optical densities on the Beer-Lambert-Bouguer law 21
6e. Effect of the environment on the absorption spectra 21
6f Absorption spectroscopy and electron transfer mechanism
in proteins 27
6g. Second - derivative absorption spectroscopy 40
6gl. Theory 40
6g2. Binding of progesterone to a^-acid glycoprotein 47

2. Fluorescence : Principles and observables

1. Introduction 55
2. Fluorescence properties 58
2a. Stokes shift 58
2b. Relation between emission spectrum and excitation wavelength 60
2c. Relation between the fluorescence intensity and the optical
density 62
2d. Fluorescence excitation spectrum 66
2e. The mirror-image rule 69
2f Fluorescence lifetime 70
2fl. Definition of the fluorescence lifetime 70
2f2. Mean fluorescence lifetime 72
2f3. Fluorescence lifetime measurement 73
viii Contents

2f4. Time Correlated Single Photon counting 79

2f5. The Strobe technique 84
2f6. Excitation with continuous light: the phase and demodulation method 85
Principles 85
Multifrequency and cross-correlation 88
2g. Fluorescence quantum yield 91
2h. Fluorescence and light diffusion 98

3. Fluorophores : Descriptions and properties

1. Introduction 99
2. Types of fluorophores 99
2a. Intrinsic fluorophores 99
2al. The aromatic amino acids 99
2a2. The Co-factors 111
2b. Extrinsincfluorophores 113
2bl. Fluorescein and rhodamine 113
2b2. Naphthalene sulfonate 114
2b3. Nucleic bases 123
2b4. Ions detectors 124
2b5. Carbohydrates fluorescent probes 127
2b6. Oxydation oftryptophan residues with N-bromosuccinimide 129
2b7. Nitration oftyrosine residues with tetranitromethane (TNM) 131
3. Effect of the environment on the fluorescence observables 134
3a. Polarity effect on the quantum yield and the position of the
emission maximum 134
3b. Effect of the viscosity on the fluorescence emission spectrum 136
3c. Effect of the environment on the fluorescence lifetime 139
3d. Relation between fluorescence and a specific sequence
in a protein 140

4. Fluorescence quenching
1. Introduction 141
2. Collisional quenching: the Stern-Volmer relation 141
3. The different types of dynamic quenching 145
4. Static quenching 158
4a. Theory 158
4b. Cytochrome c - cytochrome b2 core interaction 161
4c. Cytochrome b2 core - flavodehydrogenase interaction 162
4d. Determination of drug binding to a; - acid glycoprotein 164
5. Comparison between dynamics and static quenching 166
6. Combination of dynamic and static quenching 169
7. Thermal intensity quenching 171
8. Photoquenching 191
 Contents ix

5. Fluorescence polarization
1. Aim and definition 193
2. Principles of polarization or of photoselection 194
3. Absorption transitions and excitation polarization spectrum 197
4. Fluorescence depolarization 199
4a. Principles and applications 199
4b. Measurements of rotational correlation time oftyrosine
in smallpeptides 206
4bl. Fluorescence lifetime 206
4b2. Fluorescence intensity quenching oftyrosine residues by iodide 206
4b3. Quenching emission anisotropy 208
4c. DNA-Protein interaction 209
5. Fluorescence anisotropy decay time 211
6. Depolarization and energy transfer 214

6. Forster energy transfer

1. Principles 217
2. Energy transfer parameters 224
3. Relation between energy transfer and static quenching 227

7. Origin of protein fluorescence 237

8. Description of the structure and dynamics of a j - acid glycoprotein by

fluorescence studies
1. Introduction 261
2. Methods 266
3. Fluorescence properties of TNS bound to sialylated a r a c i d glycoprotein 267
3a. Binding parameters 267
3b. Fluorescence lifetime 270
3c. Dynamics of TNS bound to aj-acid glycoprotein 270
4. Fluorescence properties of calcofluor bound to a r a c i d glycoprotein 274
4a. Fluorescence parameters of calcofluor bound to a^-acid glycoprotein 274
4b. Binding parameters 216
4c. Nature of the interaction of calcofluor with (Xj -acid glycoprotein and HSA 277
4d. Titration of carbohydrate residues with calcofluor 280
4e. Fluorescence lifetime of calcofluor bound to a^-acid glycoprotein 285
4f Binding parameters of calcofluor white - aj-acid glycoprotein
by following the fluorescence lifetime of calcofluor 287
4g. Dynamics of calcofluor bound to a^-acid glycoprotein at equimolar
concentrations 288
4gl. Dynamics of calcofluor bound to sialylated aracid glycoprotein 288
4g2. Dynamics of Calcofluor bound to asialylated CC; - acid glycoprotein 289
x Contents

4h. Dynamics ofcalcofluor bound to aj-acid glycoprotein at excess

concentration ofcalcofluor 291
5. Fluorescence properties of the Trp residues in a r a c i d glycoprotein 293
5a. Fluorescence spectral properties 293
5b. Effect of high calcofluor concentration on the local structure of
aracid glycoprotein 296
5c. Deconvolution of the emission spectra obtained at low and high
concentrations ofcalcofluor into different components 297
5cl. Analysis as a sum of Gaussian bands 297
5c2. The Ln-normal analysis 299
6. Forster energy transfer experiments from Trp residues to calcofluor white 305
7. Relation between the secondary structure of carbohydrate residues of a r a c i d
glycoprotein and the fluorescence of the Trp residues of the protein 309
8. Effect of the secondary structure of carbohydrate residues of a r a c i d
glycoprotein on the local dynamics of the protein 313
8a. Fluorescence emission intensity of aj-acid glycoprotein
as a function of temperature 313
8b. Fluorescence emission anisotropy of aj-acid glycoprotein
as a function of temperature 315
9. Tertiary structure of a r a c i d glycoprotein : first model describing the
presence of a pocket 319
10. Are there any alternative fluorescence methods other than the QREA
or the Weber's method to put into evidence the presence of a pocket
within a r a c i d glycoprotein? 323
11. Experiments giving proofs of the presence of a pocket within
a r a c i d glycoprotein 324
77a. Binding of progesterone to aracid glycoprotein 324
lib. Binding ofhemin to a,j- acid glycoprotein 327
12. Homology modeling of a r acid glycoprotein 327
13. Dynamics of Trp residues in crystals of human a r acid glycoprotein 332
13a. Introduction 332
13b. Protein Preparation 332
13c. Fluorescence Excitation and Emission Spectra 334
13d. Dynamics of the Microenvironments of the hydrophobic Trp Residues 336
14. Structural studies of human aracid glycoprotein followed by X-rays
scattering and transmission electron microscopy. 339
14a. Small angle diffraction studies (SAXS) 339
14b. Wide angle diffraction studies (WAXS) 341
7467. Carbohydrate residues studies 341
14c. Electron microscopy studies 342

9. Structure and dynamics of hemoglobin subunits and of myoglobin

1. Introduction 345
2. Dynamics of Trp residues in hemoglobin and in its subunits 346
 Contents xi

3. Properties of protoporphyrin IX in different solvents and in apomyoglobin 349

3a. Chemical structure ofthe porphyrins 349
3b. Spectral properties ofproporphyrin IX in different solvents 351
3c. Spectral properties of protoporphyrin IX bound to apomyoglobin (MbdesFe) 357
4. Dynamics of protoporphyrin IX embedded in the heme pocket 359
4a. Protein rotational correlation time 359
4b. Activation energy of the porphyrin motions in the heme-pocket 359
4c. Residual internal motions of porphyrin 360
4d. Effect of metal on the porphyrin dynamics 362
5. Dynamics of the protein matrix and the heme pocket 364
6. Significance of the upward curvature 367
7. Effect of sucrose on the bimolecular diffusion constant 370

10. Fluorescence fingerprints of animal and vegetal species and / or varieties

1. Fluorescence fingerprints of Eisenia fetida and Eisenia andrei 373
la. Introduction 373
lb. Results 374
2. Structural characterization of varieties of crops among a species and of
genetically modified organisms: a fluorescence study 377
2a. Pioneering work 111
2b. Structural characterization of crops 379

References 387

Index 409
This page is intentionally left blank
 1

Chapter 1

LIGHT ABSORPTION BY A MOLECULE

1. Jablonski diagram or diagram of electronic transitions

Absorption of light (photons) by a population of molecules induces the passage of

electrons from the single ground electronic level So to an excited state Sn (n > 1). In the
excited state, a molecule is energetically unstable and thus it should return to the ground
state S o . This will be achieved according to two successive steps:
- The molecule at the excited state Sn will dissipate a part of its energy in the
surrounding environment reaching by that the lowest excited state Si.
- From the excited state Si, the molecule will attain the ground state So via different
competitive processes:
a) Emission of a photon with a radiative rate constant kr. Emission of a photon is called
fluorescence.
b) The energy absorbed by the molecules is dissipated in the medium as heat. This type
of energy is non radiative and occurs with a rate constant k;.
c) The excited molecules can give up their energy to molecules located near by. This
energy transfer occurs with a rate constant kq (collisional quenching), or with a rate
constant kt (energy transfer at distance).
d) A transient passage occurs to the excited triplet state T| of energy lower than Si. with
a rate constant kjsc. The triplet state is energetically unstable. Therefore, deexcitation of
the molecule will occur via different competitive phenomena:
- Emission of a photon with a rate constant kp. This phenomenon is called
phosphorescence.
- Dissipation of non radiative energy with a rate constant k'j.
- Transfer of energy to another molecule at distance (rate constant k't) or by collision
(rate constant k' q ) (Figure 1.1).
The S -> S or the T -> T transitions are called internal conversion.
The Si --> Ti transition is called intersystem crossing. For each excited state S, an
excited state T does exist.
Molecules that absorb photons are called chromophores and a chromophore that
emits a photon is called a fluorophore. Therefore, a fluorophore is inevitably a
chromophore, however a chromophore is not necessarily a fluorophore. For example,
heme does absorb light however it does not fluoresce. The absence of fluorescence is
the result of total energy transfer from the porphyrin ring to the iron. Other metals such
as zinc or tin do not quench completely porphyrin fluorescence. Thus, zincporphyrin
and tinporphyrin display fluorescence of energy lower than that of porphyrin.
It is important to indicate that thermal activation of a molecule can induce its passage
from the ground to the excited state.
2 Structure and Dynamics of Macromolecules

Figure 1.1. Jablonski diagram or the electronic transitions diagram. (Adapted from Jablonski, A,
1935.Z. Phys. 94, 38-64).
Used with permission. ©Thomas G. Chasteen, Sam Houston State University

2. Singlet and triplet states

Transitions described in Jablonski diagram take place within chemical molecules and
therefore they concern the atoms that constitute these molecules. The electrons of these
molecules are responsible for the different transitions shown in the Jablonski diagram.
For this reason, the Jablonski diagram is also called the electronic transitions diagram.
Since localization of an electron is difficult, one referred to four quantum numbers (n,
1, m and s) to characterize an electron and to differentiate it from the others. The
principal quantum number n determines the energy of any one-electron atom of nuclear
charge Z. n can assume any positive integral value, excluding zero.
The angular - momentum quantum number 1 determines the angular momentum of
the electron. It may assume all integral values from 0 to n-1 inclusive.
The magnetic quantum number m characterizes the magnetic field generated by the
electric current of the electron, circulating in a loop, m can assume all integral values
between —1 and +1 including zero.
The spin quantum number s is the result of the electron spinning about its own axis.
Thus, a local small magnet is generated with a spin s.
Singlet and triplet states depend on the quantum number of spin of the electron (s).
According to Pauli Exclusion Principle, two electrons within a defined orbital cannot
have equal four quantum numbers and thus will differ by the spin number. Two spin
numbers are attributed to an electron, + V2 and — V2. Therefore, two electrons that belong
to the same orbital will have opposite spin.
 Light Absorption by a Molecule 3

A parameter called the Multiplicity (M) is defined as being equal to:

M = | s , + s2l + 1 (1.1)

When the spins are parallel, M will be equal to 1 and we shall have the singlet state
S.
When the spins are anti-parallel, M will be equal to 3. We are in presence of a triplet
state T (Figure 1.2).

Figure 1.2. Spin configurations of the singlet and triplet states.

Upon excitation, one electron absorbs light energy and goes to an upper vacant
orbital. The molecule is in the excited state Sn. After desexcitation, the electron will
return to its original orbital and the molecule will be in the ground state So.
In the ground state, all molecules except oxygen are in the singlet form. Oxygen is in
the triplet state, and when excited it reaches the singlet state. The singlet excited state of
oxygen is destructive for cells attained by tumors. Photodynamic therapy is a process in
which a light-responsive chemical, when exposed to the appropriate wavelength of
light, is activated to undergo either a photophysical process or to initiate
photochemistry, producing molecular species which can interact with biological targets
(photosensitization).
4 Structure and Dynamics of Macromolecules

Such interactions can be exploited for biomedical applications or for basic studies.
Photosensitization is exploited for the destruction of tumors and certain non-neoplastic
target tissues in an approach termed photodynamic therapy (PDT). Compounds such as
porphyrins are localized in target cells and tissues. Upon light activation, energy
transfer occurs from porphyrins to oxygen molecules inducing by that a triplet -> singlet
transition within the oxygen state. Cells attained by tumor attract excited oxygen
molecules and then are being destroyed by these same oxygen molecules. The history of
photodetection and photodynamic therapy has been described by Ackroyd et al. (2001).
Administrated drugs can also be used instead of porphyrins to induce singlet oxygen
formation in cells.
Perotti et al (2004) studied the efficacy of 5-aminolevulinic acid (ALA) derivatives as
pro-photosensitising agent. The authors used cell line LM3 for their studies. ALA is a
precursor of protoporphyrin IX and thus injection of ALA and / or of its derivatives
into cells will induce the formation of protoporphyrin IX, a necessary molecule to
induce singlet oxygen and thus cell death.
Figure 1.3 describes porphirin synthesis from ALA and its derivatives. One can notice
that synthesis of porphyrin does not follow the same rule for all the coupounds used.

Figure 1.3. Porphyrin synthesis from 5-aminolevulinic acid (ALA) and ALA derivatives, hexyl-
ALA (He-ALA), undecanoyl-ALA and R,S-2-(hydroximethyl)tetrahydropyranyl-ALA (THP-
ALA). Cells were incubated for 3 hours in the presence of different amounts of ALA or its
derivatives. Intracellular porphyrins were determined fluorometrically and relativised per
number of cells present at the beginning of the experiment. Source: Perotti C, Fukuda H,
DiVenosa G, MacRobert AJ, Batlle A and Casas A, 2004, Br. J. Cancer. 90, 1660-1665.
Authorization of reprint accorded by Nature Publishing Group.
 Light Absorption by a Molecule 5

Figure 1.4 displays dark and PDT toxicities for ALA and its derivative. Undecanoyl-
ALA is intrinsically toxic to cells even in the absence of light (Panel A). Increasing
light doses accelerates cell death (Panels B and C).

Figure 1.4. Dark and PDT toxicity of ALA and ALA derivatives. Cells were incubated for 3
hours in the presence of different amounts of ALA or its derivatives in the dark (panel A), or
exposed to 0.25 (panel B) and 0.4 J. cm" of light (panel C). Cell survival is expressed as a
percentage of the control nonirradiated and exposed to ALA or derivatives. Source: Perotti C,
Fukuda H, DiVenosa G, MacRobert AJ, Batlle A and Casas A, 2004, Br. J. Cancer. 90, 1660-
1665. Authorization of reprint accorded by Nature Publishing Group.

Finally, the authors calculated tumor porphyrin synthesis after administration of

equimolar concentrations of ALA and ALA derivatives to mice. They found that ALA
is the best inducer of porhyrin synthesis (15.9 + 1.12 nmol/g of tissue), followed by
THP-ALA (8.83 0.95 nmol/g of tissue), He-ALA (3.11 0.42 nmol/g of tissue) and
undecanoyl-ALA (2.64 + 0.23 nmol/g of tissue). The values were calculated after three
hours of ALA or ALA derivative administration to the cell lines.
6 Structure and Dynamics of Macromolecules

The natural non-toxic hypericin is a photosensitizing anti-cancer drug. In fact,

neoplastic cells in culture respond to hypericin in a dose-dependent fashion: high doses
of light and high concentrations of photosensitizer cause apoptosis or cell necrosis,
whereas low levels of activation induce only a "stress response' that involves the
synthesis of proteins known as stress-induced or heat shock proteins (HSP) (Varriale et
al. 2002 and other cited references). The main function of HSP is to afford additional
protection to the cell as the need arises. Figure 1.5 displays hypericin uptake, release
and sub-cellular distribution in HEC1-B cells. One can notice that hypericin uptake
remained quite steady from 3 to 20 h (insert a). The sub-cellular distribution of
hypericin obtained by using ultracentrifuge-based methods was verified by checking
for the specific marker proteins c-Myc (nuclear), p-Bip (ER), actin (cytosol) and VDAC
(mitochondria) within each sub-fraction ( Fig. 1.5. insert b). The amount of hypericin in
sub-cellular fractions was estimated fluorimetrically (emission spectra) and expressed
after normalization (Fig. 1.5). Sensitization of hypericin in the nuclear fraction induces
locally DNA damage. The authors found that hypericin sensitization induces damage
that causes apoptosis or cell necrosis according to the energy adsorbed. Also, they show
that mild pre-sensitization endows cells with an unexpected high degree of photo-
tolerance, enhances HSP70 synthesis, sequentially promotes the expression of specific
apoptosis-related proteins and causes cell cycle arrest (Varriale et al. 2002).

Figure 1.5. Normalized fluorescence emission spectra (550-650 nm) of hypericin from
individual sub-cellular compartments of hypericin-loaded HEC1-B cells, a: Relative
fluorimetric estimate of cell-trapped drug at indicated times. The arrows indicate the residual
hypericin fluorescence at 3 and 20 h. Hypericin was measured fluorimetrically in three
experiments and expressed as percent ) with respect to hypericin fluorescence obtained at
time 0 (Xex= 490 nm). b: Sub-cellular fractionation was validated by Western blot using protein
markers specific for each fraction: actin (cytosol), c-Myc (nucleus), VDAC (mitochondria) and
p-Bip (membranes). Source: Varriale, L., Coppola, E., Quarto, M , Veneziani, B. M , and
Palumbo, G. 2002, FEBS Letters, 512, 287-290.
 Light Absorption by a Molecule 7

3. Forbidden and non forbidden transitions

Upon light absorption, electrons are under the effect of electric and magnetic
components. The contributions of these two components in the transition process could
be defined as:

F = (eE) + ( e v H / c ) (1.2)

where e, c, v, E and H are respectively the charge of the electron, the velocity of light,
the speed of rotation of the electron on itself, the electric and magnetic components of
the light wave. The products (E e) and (e v H / c) are respectively the contribution of
the electric and magnetic fields to the absorption phenomenon.
The speed of rotation of the electron on itself is very weak compared to the velocity
of light, thus the magnetic contribution to the absorption is negligible compared to the
electric one. Therefore, upon absorption, electronic transitions are the result of the
interaction between the electron and the electric field. For this reason, during
absorption, the displaced electron preserves the same spin orientation. This is why, only
the So -> Sn transitions are allowed and the So -> Tn transitions are forbidden.
The Jablonski diagram describes very well the phenomena that are occurring after
light absorption by a molecule. Understanding these phenomena helps to master
absorption and fluorescence methods and to overcome the major problems that a
researcher meets during his work.

4. Reading the Jablonski diagram

In the present paragraph, we are simply going to enumerate the most important
conclusions we can draw from the Jablonski diagram. Details are given in the different
corresponding sections.
a. Absorption allows the passage of the chromophore from the ground state to an
excited state, therefore absorption of photons is synonymous with excitation. This
feature allows us to compare the absorption and the excitation spectra of a molecule one
with each other. This comparison is possible for fluorophores and not for molecules that
absorb only.
b. Energy of absorption is higher than energy of emission.
c. Many phenomena other than fluorescence emission contribute to the deexcitation
of the fluorophore. These other alternatives to fluorescence are, radiationless loss,
phosphorescence, photooxidation and energy transfer. Thus, the weaker will be the
competitive phenomena, the highest will be the deexcitation via fluorescence.
d. Dissipation of energy via the non-radiative process has to be controlled if we want
to compare absorption or emission spectra at stable temperature.
e. Emission occurs from the excited state Si, independently of the excitation
wavelength. Therefore, the energy of the emission of a fluorophore would be
independent of the excitation wavelength.
f. Fluorescence energy is higher than that of phosphorescence.
g. Absorption and fluorescence do not require any reorientation of spin. However, the
inter-system crossing and phosphorescence require a spin reorientation. Therefore,
absorption and fluorescence are much faster than phosphorescence. Absorption occurs
Random documents with unrelated
content Scribd suggests to you:
 Microbiology - Concept Map
Summer 2024 - Center

Prepared by: Instructor Davis

Date: July 28, 2025

Results 1: Research findings and conclusions

Learning Objective 1: Learning outcomes and objectives
• Comparative analysis and synthesis
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Formula: [Mathematical expression or equation]
Learning Objective 2: Key terms and definitions
• Comparative analysis and synthesis
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Learning Objective 3: Theoretical framework and methodology
• Learning outcomes and objectives
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Learning Objective 4: Study tips and learning strategies
• Experimental procedures and results
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Learning Objective 5: Case studies and real-world applications
• Comparative analysis and synthesis
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Formula: [Mathematical expression or equation]
Note: Fundamental concepts and principles
• Experimental procedures and results
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Example 6: Case studies and real-world applications
• Literature review and discussion
- Sub-point: Additional details and explanations
- Example: Practical application scenario
Formula: [Mathematical expression or equation]
Remember: Study tips and learning strategies
• Historical development and evolution
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
[Figure 8: Diagram/Chart/Graph]
Key Concept: Statistical analysis and interpretation
• Experimental procedures and results
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Formula: [Mathematical expression or equation]
[Figure 9: Diagram/Chart/Graph]
Definition: Key terms and definitions
• Research findings and conclusions
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Formula: [Mathematical expression or equation]
Test 2: Best practices and recommendations
Note: Literature review and discussion
• Literature review and discussion
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Definition: Study tips and learning strategies
• Statistical analysis and interpretation
- Sub-point: Additional details and explanations
- Example: Practical application scenario
Practice Problem 12: Interdisciplinary approaches
• Problem-solving strategies and techniques
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Formula: [Mathematical expression or equation]
[Figure 13: Diagram/Chart/Graph]
Note: Comparative analysis and synthesis
• Research findings and conclusions
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Key Concept: Historical development and evolution
• Research findings and conclusions
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Definition: Comparative analysis and synthesis
• Problem-solving strategies and techniques
- Sub-point: Additional details and explanations
- Example: Practical application scenario
[Figure 16: Diagram/Chart/Graph]
Note: Critical analysis and evaluation
• Best practices and recommendations
- Sub-point: Additional details and explanations
- Example: Practical application scenario
Definition: Study tips and learning strategies
• Practical applications and examples
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
[Figure 18: Diagram/Chart/Graph]
Key Concept: Ethical considerations and implications
• Critical analysis and evaluation
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Note: Study tips and learning strategies
• Literature review and discussion
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Results 3: Ethical considerations and implications
Example 20: Key terms and definitions
• Practical applications and examples
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
[Figure 21: Diagram/Chart/Graph]
Remember: Study tips and learning strategies
• Current trends and future directions
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Formula: [Mathematical expression or equation]
Note: Key terms and definitions
• Experimental procedures and results
- Sub-point: Additional details and explanations
- Example: Practical application scenario
Important: Problem-solving strategies and techniques
• Historical development and evolution
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Remember: Critical analysis and evaluation
• Research findings and conclusions
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
[Figure 25: Diagram/Chart/Graph]
Example 25: Critical analysis and evaluation
• Statistical analysis and interpretation
- Sub-point: Additional details and explanations
- Example: Practical application scenario
[Figure 26: Diagram/Chart/Graph]
Practice Problem 26: Experimental procedures and results
• Experimental procedures and results
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Important: Current trends and future directions
• Best practices and recommendations
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Example 28: Research findings and conclusions
• Problem-solving strategies and techniques
- Sub-point: Additional details and explanations
- Example: Practical application scenario
Note: Study tips and learning strategies
• Key terms and definitions
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Formula: [Mathematical expression or equation]
Module 4: Statistical analysis and interpretation
Example 30: Assessment criteria and rubrics
• Practical applications and examples
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Formula: [Mathematical expression or equation]
Definition: Study tips and learning strategies
• Research findings and conclusions
- Sub-point: Additional details and explanations
- Example: Practical application scenario
Remember: Ethical considerations and implications
• Statistical analysis and interpretation
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Example 33: Best practices and recommendations
• Practical applications and examples
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Formula: [Mathematical expression or equation]
Example 34: Literature review and discussion
• Critical analysis and evaluation
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Example 35: Study tips and learning strategies
• Historical development and evolution
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Formula: [Mathematical expression or equation]
[Figure 36: Diagram/Chart/Graph]
Key Concept: Critical analysis and evaluation
• Practical applications and examples
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Practice Problem 37: Literature review and discussion
• Ethical considerations and implications
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Remember: Best practices and recommendations
• Experimental procedures and results
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Example 39: Research findings and conclusions
• Experimental procedures and results
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Exercise 5: Case studies and real-world applications
Practice Problem 40: Current trends and future directions
• Experimental procedures and results
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
[Figure 41: Diagram/Chart/Graph]
Definition: Case studies and real-world applications
• Statistical analysis and interpretation
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Formula: [Mathematical expression or equation]
Note: Literature review and discussion
• Ethical considerations and implications
- Sub-point: Additional details and explanations
- Example: Practical application scenario
[Figure 43: Diagram/Chart/Graph]
Note: Current trends and future directions
• Fundamental concepts and principles
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Example 44: Learning outcomes and objectives
• Historical development and evolution
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Formula: [Mathematical expression or equation]
Example 45: Historical development and evolution
• Historical development and evolution
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Formula: [Mathematical expression or equation]
Practice Problem 46: Learning outcomes and objectives
• Research findings and conclusions
- Sub-point: Additional details and explanations
- Example: Practical application scenario
Formula: [Mathematical expression or equation]
Important: Fundamental concepts and principles
• Current trends and future directions
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Practice Problem 48: Learning outcomes and objectives
• Case studies and real-world applications
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Formula: [Mathematical expression or equation]
Practice Problem 49: Experimental procedures and results
• Current trends and future directions
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Review 6: Literature review and discussion
Important: Statistical analysis and interpretation
• Fundamental concepts and principles
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Formula: [Mathematical expression or equation]
[Figure 51: Diagram/Chart/Graph]
Example 51: Fundamental concepts and principles
• Statistical analysis and interpretation
- Sub-point: Additional details and explanations
- Example: Practical application scenario
Definition: Learning outcomes and objectives
• Practical applications and examples
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Formula: [Mathematical expression or equation]
Remember: Study tips and learning strategies
• Historical development and evolution
- Sub-point: Additional details and explanations
- Example: Practical application scenario
Formula: [Mathematical expression or equation]
Key Concept: Research findings and conclusions
• Critical analysis and evaluation
- Sub-point: Additional details and explanations
- Example: Practical application scenario
Formula: [Mathematical expression or equation]
Important: Study tips and learning strategies
• Case studies and real-world applications
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Practice Problem 56: Fundamental concepts and principles
• Study tips and learning strategies
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
[Figure 57: Diagram/Chart/Graph]
Important: Learning outcomes and objectives
• Fundamental concepts and principles
- Sub-point: Additional details and explanations
- Example: Practical application scenario
[Figure 58: Diagram/Chart/Graph]
Important: Fundamental concepts and principles
• Study tips and learning strategies
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
[Figure 59: Diagram/Chart/Graph]
Important: Best practices and recommendations
• Assessment criteria and rubrics
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Discussion 7: Case studies and real-world applications
Remember: Interdisciplinary approaches
• Historical development and evolution
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
[Figure 61: Diagram/Chart/Graph]
Note: Literature review and discussion
• Fundamental concepts and principles
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Formula: [Mathematical expression or equation]
Note: Interdisciplinary approaches
• Study tips and learning strategies
- Sub-point: Additional details and explanations
- Example: Practical application scenario
[Figure 63: Diagram/Chart/Graph]
Note: Research findings and conclusions
• Best practices and recommendations
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Note: Problem-solving strategies and techniques
• Historical development and evolution
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Formula: [Mathematical expression or equation]
Note: Problem-solving strategies and techniques
• Statistical analysis and interpretation
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Example 66: Interdisciplinary approaches
• Statistical analysis and interpretation
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Formula: [Mathematical expression or equation]
[Figure 67: Diagram/Chart/Graph]
Key Concept: Research findings and conclusions
• Case studies and real-world applications
- Sub-point: Additional details and explanations
- Example: Practical application scenario
Formula: [Mathematical expression or equation]
Important: Learning outcomes and objectives
• Key terms and definitions
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Formula: [Mathematical expression or equation]
[Figure 69: Diagram/Chart/Graph]
Note: Key terms and definitions
• Ethical considerations and implications
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Formula: [Mathematical expression or equation]
Introduction 8: Problem-solving strategies and techniques
Important: Current trends and future directions
• Comparative analysis and synthesis
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Formula: [Mathematical expression or equation]
Practice Problem 71: Comparative analysis and synthesis
• Learning outcomes and objectives
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Formula: [Mathematical expression or equation]
[Figure 72: Diagram/Chart/Graph]
Key Concept: Problem-solving strategies and techniques
• Assessment criteria and rubrics
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Formula: [Mathematical expression or equation]
Remember: Assessment criteria and rubrics
• Current trends and future directions
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Key Concept: Fundamental concepts and principles
• Comparative analysis and synthesis
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Formula: [Mathematical expression or equation]
Key Concept: Research findings and conclusions
• Theoretical framework and methodology
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Example 76: Research findings and conclusions
• Comparative analysis and synthesis
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Practice Problem 77: Literature review and discussion
• Current trends and future directions
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Formula: [Mathematical expression or equation]
[Figure 78: Diagram/Chart/Graph]
Note: Study tips and learning strategies
• Assessment criteria and rubrics
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Practice Problem 79: Experimental procedures and results
• Practical applications and examples
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Conclusion 9: Assessment criteria and rubrics
Definition: Practical applications and examples
• Learning outcomes and objectives
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Important: Practical applications and examples
• Ethical considerations and implications
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Formula: [Mathematical expression or equation]
Important: Current trends and future directions
• Best practices and recommendations
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Important: Historical development and evolution
• Fundamental concepts and principles
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Practice Problem 84: Critical analysis and evaluation
• Case studies and real-world applications
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Remember: Historical development and evolution
• Best practices and recommendations
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Formula: [Mathematical expression or equation]
Key Concept: Key terms and definitions
• Key terms and definitions
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
[Figure 87: Diagram/Chart/Graph]
Example 87: Best practices and recommendations
• Experimental procedures and results
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Formula: [Mathematical expression or equation]
[Figure 88: Diagram/Chart/Graph]
Example 88: Theoretical framework and methodology
• Experimental procedures and results
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Example 89: Current trends and future directions
• Comparative analysis and synthesis
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Practice 10: Theoretical framework and methodology
Note: Ethical considerations and implications
• Experimental procedures and results
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Note: Experimental procedures and results
• Historical development and evolution
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Note: Theoretical framework and methodology
• Experimental procedures and results
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Example 93: Critical analysis and evaluation
• Research findings and conclusions
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Remember: Case studies and real-world applications
• Best practices and recommendations
- Sub-point: Additional details and explanations
- Example: Practical application scenario
Practice Problem 95: Fundamental concepts and principles
• Learning outcomes and objectives
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Definition: Current trends and future directions
• Learning outcomes and objectives
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Formula: [Mathematical expression or equation]
Practice Problem 97: Assessment criteria and rubrics
• Best practices and recommendations
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Example 98: Learning outcomes and objectives
• Problem-solving strategies and techniques
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Example 99: Assessment criteria and rubrics
• Best practices and recommendations
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
[Figure 100: Diagram/Chart/Graph]
References 11: Research findings and conclusions
Definition: Case studies and real-world applications
• Statistical analysis and interpretation
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Practice Problem 101: Ethical considerations and implications
• Historical development and evolution
- Sub-point: Additional details and explanations
- Example: Practical application scenario
[Figure 102: Diagram/Chart/Graph]
Note: Theoretical framework and methodology
• Statistical analysis and interpretation
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Formula: [Mathematical expression or equation]
[Figure 103: Diagram/Chart/Graph]
Practice Problem 103: Historical development and evolution
• Critical analysis and evaluation
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Formula: [Mathematical expression or equation]
[Figure 104: Diagram/Chart/Graph]
Definition: Case studies and real-world applications
• Ethical considerations and implications
- Sub-point: Additional details and explanations
- Example: Practical application scenario
[Figure 105: Diagram/Chart/Graph]
Remember: Experimental procedures and results
• Problem-solving strategies and techniques
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Formula: [Mathematical expression or equation]
Important: Fundamental concepts and principles
• Interdisciplinary approaches
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Formula: [Mathematical expression or equation]
Example 107: Critical analysis and evaluation
• Practical applications and examples
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Formula: [Mathematical expression or equation]
Key Concept: Comparative analysis and synthesis
• Assessment criteria and rubrics
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
[Figure 109: Diagram/Chart/Graph]
Example 109: Study tips and learning strategies
• Comparative analysis and synthesis
- Sub-point: Additional details and explanations
- Example: Practical application scenario
[Figure 110: Diagram/Chart/Graph]
Quiz 12: Study tips and learning strategies
Practice Problem 110: Case studies and real-world applications
• Case studies and real-world applications
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Remember: Ethical considerations and implications
• Theoretical framework and methodology
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Note: Research findings and conclusions
• Ethical considerations and implications
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Important: Learning outcomes and objectives
• Experimental procedures and results
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Formula: [Mathematical expression or equation]
Important: Theoretical framework and methodology
• Assessment criteria and rubrics
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Formula: [Mathematical expression or equation]
[Figure 115: Diagram/Chart/Graph]
Note: Comparative analysis and synthesis
• Case studies and real-world applications
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Remember: Learning outcomes and objectives
• Critical analysis and evaluation
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Remember: Theoretical framework and methodology
• Critical analysis and evaluation
- Sub-point: Additional details and explanations
- Example: Practical application scenario
[Figure 118: Diagram/Chart/Graph]
Example 118: Study tips and learning strategies
• Key terms and definitions
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Remember: Study tips and learning strategies
• Research findings and conclusions
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Exercise 13: Fundamental concepts and principles
Important: Literature review and discussion
• Fundamental concepts and principles
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Formula: [Mathematical expression or equation]
Important: Learning outcomes and objectives
• Interdisciplinary approaches
- Sub-point: Additional details and explanations
- Example: Practical application scenario
Formula: [Mathematical expression or equation]
Practice Problem 122: Research findings and conclusions
• Best practices and recommendations
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Important: Key terms and definitions
• Learning outcomes and objectives
- Sub-point: Additional details and explanations
- Example: Practical application scenario
Formula: [Mathematical expression or equation]
Definition: Statistical analysis and interpretation
• Study tips and learning strategies
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Formula: [Mathematical expression or equation]
[Figure 125: Diagram/Chart/Graph]
Example 125: Historical development and evolution
• Theoretical framework and methodology
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Practice Problem 126: Learning outcomes and objectives
• Practical applications and examples
- Sub-point: Additional details and explanations
- Example: Practical application scenario
- Note: Important consideration
Formula: [Mathematical expression or equation]
Welcome to our website – the ideal destination for book lovers and
knowledge seekers. With a mission to inspire endlessly, we offer a
vast collection of books, ranging from classic literary works to
specialized publications, self-development books, and children's
literature. Each book is a new journey of discovery, expanding
knowledge and enriching the soul of the reade

Our website is not just a platform for buying books, but a bridge
connecting readers to the timeless values of culture and wisdom. With
an elegant, user-friendly interface and an intelligent search system,
we are committed to providing a quick and convenient shopping
experience. Additionally, our special promotions and home delivery
services ensure that you save time and fully enjoy the joy of reading.

Let us accompany you on the journey of exploring knowledge and

personal growth!

ebookfinal.com