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Experimental and stimulation analyses of fluorescent solvent relaxation process in biomembranes : Inflence of ions and molecular interpretation of the dye dynamics / Analyse expérimentale et numérique des processus de relaxation de solvant dans une membrane biologique : Rôle des ions et interprétation moléculaire de la dynamique des marqueurs fluorescentsBarucha-Kraszewska, Justyna 31 October 2012 (has links)
De nombreux processus biologiques liés aux membranes cellulaires lipidiques sont encore très mal connus. La présence d'eau et d'ions à l'interface influence les propriétés structurelles et dynamiques de la bicouche lipidique. Les techniques de fluorescence sont très utiles pour étudier les membranes en raison de la grande sensibilité des sondes à leur environnement. Nous avons utilisé la technique de relaxation de solvant (SR) pour explorer l'hydratation et la mobilité de l'eau. Nous avons également réalisé des calculs quantiques (QM) et des dynamiques moléculaires (DM) pour étayer nos expériences. Les résultats SR montrent qu'un petit cation (Na+) est très attiré par la membrane et augmente sa rigidité à l'opposé des cations (NH4+, Cs+) plus gros. Les anions (CI04-, SCN-) s'adsorbent à l'interface plus facilement que Cl-. Ces anions changent la mobilité et l'hydratation des têtes polaires des lipides de la bicouche. Les études SR de la zone hydrophobe de la membrane montrent que les processus de relaxation sont ici très complexes. lis reflètent des processus rapides intramoléculaire (relaxation de torsion, transferts de charge) et des processus intermoléculaires lents. Les calculs QM ont permis de créer les champs de force de trois sondes fluorescentes (Prodan, Laurdan et C-laurdan). Les simulations DM ont permis de déterminer les positions des sondes dans une membrane DOPC. La modélisation reproduit correctement les résultats SR, en particulier les temps de relaxation : de l'ordre de la ps en solvant aqueux et de la ns dans la membrane. Les simulations MD sont complémentaires des méthodes SR et permettent de surveiller le comportement de molécules uniques. / Many biologically important processes and phcnomena in lipid membranes are still not fully understood. The presence of ions and water molœules has a significant influence on the structural and dynamical properties of lipid bilayers. Fluorescent techniques are versatile tools for studying the lipid membranes, because the fluorescence emission is strongly sensitive to dye environment. We have conducted fluorescent solvent relaxation (SR) experiments to explore the hydration and mobility properties in lipid membranes in the presence of different chaotropic ions. We have also carried out Quantum Mechanical (QM) calculations and Molecular Dynamics (MD) simulations for supporting the SR experiments. SR experiments show that small cation (Na+) is attracted to the membrane and increases rigidity ofbilayer, while larger cations (NH/, Cs+) should not. Large anions (CI04·, SCN') adsorl, at the membrane interface more easily than smaller ones (Cl') and significantly change tl!e mobility and hydration of the headgroup region oflipid bilayer. SR study ofhydrophobic part of the membrane show that SR processes are complex there and reflect botl!: faster, intramolecular (torsional relaxation or fonnation of charge transfer state) and slower, intermolecular (SR) relaxation processes. QM calculatiom were used to create force-field for three fluorescent dyes (Prodan, Laurdan and C-laurdan). MD simulations allow detennining position of the dye in the lipid membrane in the ground state and after excitation and reproduce correctly SR timescale- ps in water and ns in the membrane. MD simulations extend the capabilities of SR method and allow observing the behaviour of individual molecules.
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Unravelling the Interaction of DNA Origami with Chaotropic Agents: Anion-Specific Stability and Water-Driven EffectsDornbusch, Daniel 01 August 2024 (has links)
In dieser Arbeit werden systematisch die bisher unerforschten grundlegenden physikalischen und chemischen Eigenschaften von DNA-Origami untersucht, die die Stabilität dieser aus doppelsträngiger DNA aufgebauten nanoskopischen Suprastrukturen bestimmen. In Analogie zu den zahlreichen Studien, die sich mit der Stabilität von Proteinen durch kontrollierte Denaturierung beschäftigen, spielen auch in dieser Arbeit die Denaturierungsbedingungen eine zentrale Rolle. Unter Verwendung von Guanidinium (Gdm+) als teilweise DNA-stabilisierendes, aber auch potentiell denaturierendes Kation steht dessen Wirkung auf DNA-Origami-Dreiecke im Mittelpunkt der Untersuchungen, wobei insbesondere die unerwartete Modulation der nanoskopischen Schädigung von DNA-Origami durch die begleitenden Gegenanionen zu Gdm+ im Vordergrund steht. Die Experimente zielen darauf ab, atomistische, molekulare, nanoskopische und thermodynamische Eigenschaften von DNA-Origami zu korrelieren und zu klären, wie diese vom Design des DNA-Origami selbst abhängen können.
Die Ergebnisse zeigen einen unerwarteten Zusammenhang zwischen den spezifischen Gegenanionen des Denaturierungsmittels und der Stabilität der DNA-Origami-Dreiecke: Sulfat wirkt stabilisierend, während Chlorid die Superstruktur bereits unterhalb der globalen Schmelztemperatur destabilisiert. Statistische Analysen von Rasterkraftmikroskop (AFM)-Bildern und Zirkulardichroismus (CD)-Spektren zeigen Strukturübergänge auf nano-skopischer bzw. molekularer Ebene. Werden diese Techniken mit thermischer Denaturierung in Gegenwart von schwacher bis starker chemischer Denaturierung kombiniert, so zeigt sich, dass Änderungen der Wärmekapazität (ΔCp) während der strukturellen Veränderungen der DNA-Originale eine Schlüsselrolle bei der Bestimmung ihrer Empfindlichkeit gegenüber Temperatur und Denaturierungsmitteln spielen. Die Daten deuten darauf hin, dass Wasser auf apolaren DNA-Origami-Oberflächen der molekulare Ursprung der abgeleiteten Wärme-kapazitätsänderungen ist. Diese Hypothese wird durch Molekulardynamik-Simulationen (MD) unterstützt, die die Modulation von ΔCp durch die Hydratationshüllen der Anionen zeigen. Ihr unterschiedliches Potential, stabile Ionenpaare mit Gdm+ in konzentrierten Salzlösungen zu bilden, kann die experimentell beobachteten Variationen der strukturellen Stabilität erklären. Die Kopplung von strukturellen Übergängen an ΔCp wird somit als Schlüsselfaktor für die Destabilisierung von DNA-Origami sowohl bei höheren als auch bei niedrigeren Temperaturen identifiziert. Darüber hinaus weisen DNA-Origami nicht nur diese Eigenschaft auf, sondern ermöglichen auch die Beobachtung von kalten Denaturierungsprozessen auf nanoskopischer Ebene, bei denen kälteinduzierte Spannungen innerhalb der Superstruktur bei einem Bruch an vorherbestimmten lokalen Stellen freigesetzt werden, die in AFM-Bildern sichtbar sind. Dies ist die erste Beobachtung der kälteinduzierten Denaturierung von Nukleinsäuren bei Temperaturen über 0 °C sowie von DNA-basierten Superstrukturen.
In dieser Arbeit wird die strukturelle Stabilität von sechs verschiedenen 2D- und 3D-DNA-Origami-Nanostrukturen in unterschiedlichen chemischen Umgebungen untersucht. Drei chaotrope Salze - Guanidiniumsulfat (Gdm2SO4), Guanidiniumchlorid (GdmCl) und Tetrapropylammoniumchlorid (TPACl) - werden als Denaturierungsmittel verwendet. Mittels Rasterkraftmikroskopie wird die Integrität der Nanostrukturen quantifiziert, wobei sich Gdm2SO4 als das schwächste und TPACl als das stärkste Denaturierungsmittel für DNA-Origami erweist, was sich auch in den Schmelztemperaturen widerspiegelt. Die Abhängigkeit der DNA-Origami-Stabilität von der Superstruktur wird besonders bei 3D-Nanostrukturen deutlich. Hier zeigen mechanisch flexible Designs sowohl in GdmCl als auch in TPACl eine höhere Stabilität als ihre starren Gegenstücke. Die Abhängigkeit der DNA-Origami-Stabilität von der Superstruktur wird besonders in 3D-Nanostrukturen deutlich, in denen mechanisch flexible Strukturen sowohl in GdmCl als auch in TPACl eine höhere Stabilität aufweisen als ihre steifen Gegenstücke. Dies begünstigt die Bildung von intramolekularen Verformungen, die sich entweder in 'weichen' Architekturen über die gesamte Superstruktur verteilen oder in ansonsten 'steifen' Strukturen in den weniger stabilen Regionen konzentrieren.:Table of contents
Questions addressed in this thesis .................................................................................... I
Abstract ................................................................................................................................ I
Englisch .................................................................................................................................................. I
Deutsch .................................................................................................................................................. II
Acronyms ........................................................................................................................... III
Substances .......................................................................................................................................... IV
Physical and Chemical abbreviations ............................................................................................... IV
Mathematical abbreviations ................................................................................................................ V
1 Introduction ................................................................................................................. 1
1.1
Deoxyribonucleic acid ................................................................................................................. 1
1.1.1 The structure of DNA ............................................................................................................ 1
1.1.2 Hydrogen bonds .................................................................................................................... 1
1.1.3 Base stacking ........................................................................................................................ 2
1.1.4 Water DNA interactions: A complex dance of stability and dynamics .................................. 5
1.1.5 The effect of ionic strength on DNA conformation ................................................................ 9
1.1.6 Conformational changes ....................................................................................................... 9
1.1.7 Forms of DNA ..................................................................................................................... 10
1.1.8 The role of apolar groups in DNA unfolding ........................................................................ 13
1.1.9 Energetics of DNA structural transitions ............................................................................. 14
1.1.10 Melting temperature ............................................................................................................ 15
1.2
Hofmeister series ....................................................................................................................... 17
1.2.1 Probing the Hofmeister series: Salt effects biomolecules ................................................... 17
1.2.2 Specific ion effects in electrolyte solutions .......................................................................... 19
1.3
DNA nanostructures................................................................................................................... 20
1.3.1 DNA origami ........................................................................................................................ 22
1.3.2 Challenges in DNA origami stability .................................................................................... 26
1.3.3 DNA origami in single molecule studies .............................................................................. 27
1.4
Circular dichroism ...................................................................................................................... 28
1.4.1 Circular dichroism spectroscopy for analyzing DNA conformations ................................... 30
1.4.2 Wavelength-dependent spectroscopic signatures of DNA conformation ........................... 32
1.5
Atomic force microscopy .......................................................................................................... 34
1.6
2D correlation spectroscopy ..................................................................................................... 36
1.6.1 2D correlation spectroscopy: Synchronous and asynchronous spectra analysis ............... 39
1.6.2 Perturbation-correlation moving-window 2D correlation spectroscopy ............................... 40
1.7
Multivariate analysis of spectral data using PCA and ITTFA ................................................ 41
1.8
Cold denaturation ....................................................................................................................... 42
2 Results and Discussion ............................................................................................ 44
2.1
Cold denaturation of the Rothemund DNA origami triangle .................................................. 45
2.2
Heat denaturation of the Rothemund DNA origami triangle .................................................. 50
2.2.1 Investigations by atomic force microscopy ......................................................................... 51
2.2.2 Circular dichroism spectroscopy and thermodynamic modelling ........................................ 56
2.2.3 Divergent effects of Cl- and SO42- on DNA origami stability ................................................ 62
2.3
Magnesium concentration modulation of DNA Origami heat denaturation ......................... 65
2.4
Assessing DNA origami stability in different chaotropic environments .............................. 66
2.4.1 DNA origami integrity influenced by Gdm2SO4 ................................................................... 67
2.4.2 DNA origami integrity influenced by GdmCl ........................................................................ 73
2.4.3 DNA origami integrity influenced by TPACl ........................................................................ 75
2.4.4 Quantitative comparison ..................................................................................................... 77
3 Critics ......................................................................................................................... 78
4 Conclusion ................................................................................................................. 79
5 Outlook ...................................................................................................................... 81
6 Material and Methods ................................................................................................ 82
6.1
DNA origami synthesis .............................................................................................................. 82
6.2
Sample preparation and AFM imaging ..................................................................................... 82
6.2.1 Anion-specific structure and stability of guanidinium-bound DNA origami & Cold denaturation of DNA origami nanostructures ...................................................................... 82
6.2.2 Superstructure-dependent stability of DNA origami nanostructures in the presence of chaotropic denaturants ........................................................................................................ 83
6.2.3 Cold denaturation of DNA origami nanostructures ............................................................. 83
6.3
CD spectroscopy and analysis ................................................................................................. 84
6.3.1 Anion-specific structure and stability of guanidinium-bound DNA origami ......................... 84
6.3.2 Pre-treatment of the CD data and calculation of melting temperatures .............................. 84
6.3.3 Cold denaturation of DNA origami nanostructures ............................................................. 84
6.3.4 Superstructure-dependent stability of DNA origami nanostructures in the presence of chaotropic denaturants ........................................................................................................ 84
6.4
Principal component analysis and iterative target test factor analysis ............................... 85
6.5
Thermodynamic modelling ........................................................................................................ 85
6.6
Molecular dynamics modelling ................................................................................................. 85
Appendix ........................................................................................................................... 88
Acknowledgment ............................................................................................................ 100
Bibliography .................................................................................................................... 101
List of Figures ................................................................................................................. 116
List of Tables ................................................................................................................... 118
Declaration of independence – Selbstständigkeitserklärung ...................................... 119 / This thesis undertakes the systematic study of hitherto unexplored fundamental physical and chemical properties of DNA origami that determine the stability of these designed nanoscopic superstructural assemblies of double-stranded DNA. In analogy to the vast number of studies addressing protein stability by controlled denaturation, denaturing conditions play a central role in this thesis as well. Using guanidinium (Gdm+) as a partly DNA-stabilizing but also potentially denaturing cation, its effect on DNA origami triangles is central to the study which particularly addressed the unexpected modulation of nanoscopic damage of DNA origami by the accompanying counter-anions to Gdm+. The experiments aim at correlating atomistic, molecular, nanoscopic and thermodynamic properties of DNA origami and at elucidating how these may depend on the DNA origami design itself.
The results demonstrate an unexpected relationship between the specific counter-anions of the denaturant and the stability of DNA origami triangles: sulphate exhibits stabilizing effects and chloride induces destabilization of the superstructure already below the global melting temperature. Statistical analyses of both atomic force microscopy (AFM) images and circular dichroism (CD) spectra reveal structural transitions at the nanoscopic and molecular level, respectively. Combining these techniques with thermal denaturation in the presence of mild to strong chemical denaturation, changes in heat capacity (ΔCp) during DNA origami structural changes are shown to play the key role in determining their sensitivity to temperature and denaturants. The data suggest that water at apolar DNA origami surfaces is the molecular origin of the derived heat capacity changes. This hypothesis is substantiated by Molecular Dynamics (MD) simulations which shed light on the modulation of ΔCp by the hydration shells of anions. Their different potential to form stable ion pairs with Gdm+ in concentrated salt solutions can explain the experimentally observed variations of structural stability.
The coupling of structural transitions to ΔCp is thus identified as a key factor in the destabilization of DNA origami at both elevated and lowered temperatures. Furthermore, DNA origami not only exhibit this property, but also enable the observation of cold denaturation processes at the nanoscopic level, where cold-induced strain within the superstructure is released upon breakage at predisposed local sites, visible in AFM images. This is the first observation of cold-induced denaturation of nucleic acids at temperatures above 0 °C, as well of DNA-based superstructures.
Extending the scope, the work evaluates the structural stability of six different 2D and 3D DNA origami nanostructures in different chemical environments. Three chaotropic salts - guanidinium sulfate (Gdm2SO4), guanidinium chloride (GdmCl), and tetrapropylammonium chloride (TPACl) - are used as denaturants. Atomic force microscopy quantifies the nanostructural integrity, revealing Gdm2SO4 as the weakest and TPACl as the strongest DNA origami denaturant, which is also reflected in the melting temperatures. The dependence of DNA origami stability on its superstructure is particularly evident in 3D nanostructures, where mechanically flexible designs exhibit higher stability in both GdmCl and TPACl than rigid counterparts. This supports the buildup of intramolecular strain, which becomes either partitioned among the entire superstructure in “soft” architectures or accumulates at the least stable regions in otherwise “rigid” designs.:Table of contents
Questions addressed in this thesis .................................................................................... I
Abstract ................................................................................................................................ I
Englisch .................................................................................................................................................. I
Deutsch .................................................................................................................................................. II
Acronyms ........................................................................................................................... III
Substances .......................................................................................................................................... IV
Physical and Chemical abbreviations ............................................................................................... IV
Mathematical abbreviations ................................................................................................................ V
1 Introduction ................................................................................................................. 1
1.1
Deoxyribonucleic acid ................................................................................................................. 1
1.1.1 The structure of DNA ............................................................................................................ 1
1.1.2 Hydrogen bonds .................................................................................................................... 1
1.1.3 Base stacking ........................................................................................................................ 2
1.1.4 Water DNA interactions: A complex dance of stability and dynamics .................................. 5
1.1.5 The effect of ionic strength on DNA conformation ................................................................ 9
1.1.6 Conformational changes ....................................................................................................... 9
1.1.7 Forms of DNA ..................................................................................................................... 10
1.1.8 The role of apolar groups in DNA unfolding ........................................................................ 13
1.1.9 Energetics of DNA structural transitions ............................................................................. 14
1.1.10 Melting temperature ............................................................................................................ 15
1.2
Hofmeister series ....................................................................................................................... 17
1.2.1 Probing the Hofmeister series: Salt effects biomolecules ................................................... 17
1.2.2 Specific ion effects in electrolyte solutions .......................................................................... 19
1.3
DNA nanostructures................................................................................................................... 20
1.3.1 DNA origami ........................................................................................................................ 22
1.3.2 Challenges in DNA origami stability .................................................................................... 26
1.3.3 DNA origami in single molecule studies .............................................................................. 27
1.4
Circular dichroism ...................................................................................................................... 28
1.4.1 Circular dichroism spectroscopy for analyzing DNA conformations ................................... 30
1.4.2 Wavelength-dependent spectroscopic signatures of DNA conformation ........................... 32
1.5
Atomic force microscopy .......................................................................................................... 34
1.6
2D correlation spectroscopy ..................................................................................................... 36
1.6.1 2D correlation spectroscopy: Synchronous and asynchronous spectra analysis ............... 39
1.6.2 Perturbation-correlation moving-window 2D correlation spectroscopy ............................... 40
1.7
Multivariate analysis of spectral data using PCA and ITTFA ................................................ 41
1.8
Cold denaturation ....................................................................................................................... 42
2 Results and Discussion ............................................................................................ 44
2.1
Cold denaturation of the Rothemund DNA origami triangle .................................................. 45
2.2
Heat denaturation of the Rothemund DNA origami triangle .................................................. 50
2.2.1 Investigations by atomic force microscopy ......................................................................... 51
2.2.2 Circular dichroism spectroscopy and thermodynamic modelling ........................................ 56
2.2.3 Divergent effects of Cl- and SO42- on DNA origami stability ................................................ 62
2.3
Magnesium concentration modulation of DNA Origami heat denaturation ......................... 65
2.4
Assessing DNA origami stability in different chaotropic environments .............................. 66
2.4.1 DNA origami integrity influenced by Gdm2SO4 ................................................................... 67
2.4.2 DNA origami integrity influenced by GdmCl ........................................................................ 73
2.4.3 DNA origami integrity influenced by TPACl ........................................................................ 75
2.4.4 Quantitative comparison ..................................................................................................... 77
3 Critics ......................................................................................................................... 78
4 Conclusion ................................................................................................................. 79
5 Outlook ...................................................................................................................... 81
6 Material and Methods ................................................................................................ 82
6.1
DNA origami synthesis .............................................................................................................. 82
6.2
Sample preparation and AFM imaging ..................................................................................... 82
6.2.1 Anion-specific structure and stability of guanidinium-bound DNA origami & Cold denaturation of DNA origami nanostructures ...................................................................... 82
6.2.2 Superstructure-dependent stability of DNA origami nanostructures in the presence of chaotropic denaturants ........................................................................................................ 83
6.2.3 Cold denaturation of DNA origami nanostructures ............................................................. 83
6.3
CD spectroscopy and analysis ................................................................................................. 84
6.3.1 Anion-specific structure and stability of guanidinium-bound DNA origami ......................... 84
6.3.2 Pre-treatment of the CD data and calculation of melting temperatures .............................. 84
6.3.3 Cold denaturation of DNA origami nanostructures ............................................................. 84
6.3.4 Superstructure-dependent stability of DNA origami nanostructures in the presence of chaotropic denaturants ........................................................................................................ 84
6.4
Principal component analysis and iterative target test factor analysis ............................... 85
6.5
Thermodynamic modelling ........................................................................................................ 85
6.6
Molecular dynamics modelling ................................................................................................. 85
Appendix ........................................................................................................................... 88
Acknowledgment ............................................................................................................ 100
Bibliography .................................................................................................................... 101
List of Figures ................................................................................................................. 116
List of Tables ................................................................................................................... 118
Declaration of independence – Selbstständigkeitserklärung ...................................... 119
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Influência de índices hematimétricos e bioquímicos de pacientes submetidas à cirurgia bariátrica sobre a estabilidade de membrana de eritrócitosArvelos, Leticia Ramos de 19 December 2014 (has links)
Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / CHAPTER II: The stability of the erythrocyte membrane, which is essential for maintenance of cell
functions, occurs in a critical region of fluidity, which depends largely on its composition and
the composition and characteristics of the medium. As the composition of the erythrocyte
membrane is influenced by several blood variables, the stability of the erythrocyte
membrane must have relations with them. The present study aimed to evaluate, by
bivariate and multivariate statistical analyses, the correlations and causal relationships
between hematological and biochemical variables and the stability of the erythrocyte
membrane against the chaotropic action of ethanol. The validity of this type of analysis
depends on the homogeneity of the population and on the variability of the studied
parameters, conditions that can be filled by patients who undergo bariatric surgery by the
technique of Roux-en-Y gastric bypass, since they will suffer feeding restrictions that have
great impact on their blood composition. Pathway analysis revealed that an increase in
hemoglobin leads to decreased stability of the cell, probably through a process mediated by
an increase in MCV. Furthermore, an increase in the MCH leads to an increase in the
erythrocyte membrane stability, probably because higher values of MCH are associated to
smaller quantities of RBC and larger contact area between the cell membrane and ethanol
present in the medium.
CHAPTER III: The need to treat obesity, a growing worldwide public health problem, has led to an
increase in performing bariatric surgery, particularly the Roux-en-Y gastric bypass. The
sudden change in eating habits, resulting from this type of surgery, leads to abrupt changes
in the body. This study analyzed the correlation between the osmotic stability of
erythrocytes and various biochemical and hematological indices in a population consisting
of 24 female volunteers, before and at four different times after surgery, distributed along
eight weeks, what allowed the generation of 120 sampling points. The osmotic stability of
erythrocytes proved to be of great importance for understanding the meaning of the redcell
distribution width (RDW), because the stability variables (1/H50 and dX) were positively
correlated with this hematological index. However, the stability variables and RDW seem to
suffer different influences from other variables, as the LDL-cholesterol (LDL-C), because only
RDW has increased throughout time. Indeed, the stability of variable 1/H50 showed positive
correlation with the blood levels of LDL-C, which declined throughout time. Path analysis
showed that the body mass index (BMI) has an indirect effect, mediated by RDW, on the
osmotic stability of erythrocytes. The correlations that the osmotic stability variables
presented with RDW may help to understand the origin of the predictive ability of this
hematological index in relation to various pathological conditions. / CAPÍTULO II: A estabilidade de membrana de eritrócitos, que é essencial para a manutenção da função
dessas células, ocorre em uma região crítica de fluidez, que depende largamente de sua
composição e das características do meio. Como a composição da membrana do eritrócito é
influenciada por muitas variáveis sanguíneas, a estabilidade de membrana do eritrócito
deve ter relações com elas. O presente estudo objetivou avaliar, por análises estatísticas
bivariadas e multivariadas, as correlações e relações causais entre variáveis hematológicas e
bioquímicas e a estabilidade de membrana de eritrócitos conta a ação caotrópica do etanol.
A validade deste tipo de análise depende da homogeneidade da população e da
variabilidade dos parâmetros estudados, condições que podem ser satisfeitas por pacientes
que sofrem cirurgia bariátrica pela técnica do desvio gástrico em Y-de-Roux, uma vez que
eles passam por restrições alimentares que têm grande impacto sobre a composição
sanguínea deles. A análise de caminho revelou que um aumento na concentração de
hemoglobina leva a uma diminuição da estabilidade da célula, provavelmente através de um
processo mediado por um aumento no volume corpuscular médio (MCV). Além disso, um
aumento na hemoglobina corpuscular media (MCH) leva a um aumento na estabilidade de
membrana do eritrócito, provavelmente porque valores elevados de MCH são associados a
menores quantidades de células vermelhas (RBC) e maiores áreas de contato entre a
membrana da célula e o etanol presente no meio.
CAPÍTULO III: A necessidade de tratar a obesidade, um crescente problema de saúde pública em todo o
mundo, tem levado a um aumento na execução de cirurgia bariátrica, particularmente o
desvio gástrico pelo Y-de-Roux. A súbita mudança nos hábitos alimentares, resultante deste
tipo de cirurgia, leva a mudanças abruptas no corpo. Este estudo analisou a correlação entre
a estabilidade osmótica de eritrócitos e vários índices hematológicos e bioquímicos em uma
população constituída de 24 participantes do sexo feminino, antes e em quatro diferentes
momentos após a cirurgia, distribuídos ao longo de oito semanas, o que permitiu a geração
de 120 pontos amostrais. A estabilidade osmótica de eritrócitos mostrou ser de grande
importância para a compreensão do significado da distribuição de volumes das células
vermelhas do sangue (RDW), porque as variáveis de estabilidade (1/H50 and dX) foram
positivamente correlacionadas com este índice hematológico. Entretanto, as variáveis de
estabilidade e o RDW parecem sofrer diferentes influências de outras variáveis, como o LDLcolesterol
(LDL-C), porque somente o RDW aumentou ao longo do tempo após a cirurgia.
Realmente, a variável de estabilidade 1/H50 apresentou correlação positiva com os níveis
sanguíneos de LDL-C, os quais diminuíram ao longo do tempo. A análise de caminho
mostrou que o índice de massa corporal (BMI) tem um efeito indireto, mediado pelo RDW,
sobre a estabilidade osmótica de eritrócitos. As correlações que as variáveis de estabilidade
osmótica apresentaram com RDW podem ajudar na compreensão da origem da habilidade
preditiva que este índice hematológico tem em relação a várias condições patológicas. / Doutor em Genética e Bioquímica
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