Insight into the Spatial Arrangement of the Lysine Tyrosylquinone and Cu2+ in the Active Site of Lysyl Oxidase-like 2

Lysyl oxidase-2 (LOXL2) is a Cu2+ and lysine tyrosylquinone (LTQ)-dependent amine oxidase that catalyzes the oxidative deamination of peptidyl lysine and hydroxylysine residues to promote crosslinking of extracellular matrix proteins. LTQ is post-translationally derived from Lys653 and Tyr689, but its biogenesis mechanism remains still elusive. A 2.4 Å Zn2+-bound precursor structure lacking LTQ (PDB:5ZE3) has become available, where Lys653 and Tyr689 are 16.6 Å apart, thus a substantial conformational rearrangement is expected to take place for LTQ biogenesis. However, we have recently shown that the overall structures of the precursor (no LTQ) and the mature (LTQ-containing) LOXL2s are very similar and disulfide bonds are conserved. In this study, we aim to gain insights into the spatial arrangement of LTQ and the active site Cu2+ in the mature LOXL2 using a recombinant LOXL2 that is inhibited by 2-hydrazinopyridine (2HP). Comparative UV-vis and resonance Raman spectroscopic studies of the 2HP-inhibited LOXL2 and the corresponding model compounds and an EPR study of the latter support that 2HP-modified LTQ serves as a tridentate ligand to the active site Cu2. We propose that LTQ resides within 2.9 Å of the active site of Cu2+ in the mature LOXL2, and both LTQ and Cu2+ are solvent-exposed

Etude des enzymes de la chaîne respiratoire caractérisées par électrochimie et spectroscopie

This thesis is focused on the study of two members of the heme-copper oxidase family by using spectroscopic and electrochemical techniques. In the first chapter cytochrome bo3 oxidase from E. coli was studied. We focused on the quinone-enzyme interactions by using quinones with different isoprenyl chains. Our aim was to better understand the role of isoprenyl chain on the catalytic activity of the enzyme and the redox properties of the heme cofactors. In the next step we studied the residues that are suggested to be in the high-affinity (QH) quinone binding site. Several site-directed mutants of these residues were investigated in order to better understand the position of QH binding site and the importance of each residue. In the last part of this chapter surface-enhanced infrared absorption spectroscopy (SEIRAS) was introduced as an alternative technique to study the membrane proteins. In the second chapter cytochrome c dependent nitric oxide reducates (cNOR) from P. denitrificans was studied. We focused on the effect of different environment (pH, proteoliposomes) on the stability of cNOR. For that purpose three pH values (6.5, 7.5 and 8.5) was selected and some of the cNOR samples were reconstituted in liposomes. Finally, the terminal proton donor (to the binuclear center) in cNOR was investigated. We studied the ligands of the Ca2+ site in cNOR since it was suggested that the proton donor may be close to this area.Le présent travail porte sur l’étude de deux protéines de la famille des oxydases à hème-fer par des techniques de spectroscopie et d’électrochimie. Le premier chapitre décrit l’étude du cytochrome bo3 oxydase issue d’E. coli. Nous nous sommes intéressés à l’étude des interactions enzyme-quinone par l’utilisation de quinones avec des longueurs chaines isoprenyl différentes. Notre but est de mieux comprendre le rôle de la longueur de la chaine des quinones sur l’activité catalytique de l’enzyme et sur les propriétés redox des cofacteurs à hème. Dans l’étape suivante, on a étudié les résidus impliqués dans le site de liaison des quinones (haute affinité, QH). Plusieurs mutations de ces résidus sont étudiées pour mieux comprendre l’importance de chacun des résidus dans cette liaison. Dans la dernière partie de ce premier chapitre, la spectroscopie SEIRAS «spectroscopie d’absorption infrarouge exaltée de surface» est introduite comme une technique alternative pour l’étude des protéines membranaires. Dans le second chapitre, la protéine membranaire cNOR issue de P. denitrificans est étudiée. Nous nous sommes focalisés sur l’effet de différents environnements (pH, présence de protéo-liposomes) sur la stabilité de la cNOR. Pour ce faire, trois valeurs de pH (6.5, 7.5 et 8.5) sont choisies et quelques échantillons de cNOR sont reconstitués dans des protéo-liposomes. Enfin, le donneur de proton terminal (au centre binucléaire) dans la protéine cNOR était étudié. De plus, nous avons étudié les ligands des ions Ca2+ puisqu’il est proposé que le donneur de proton est situé proche de cette région

Temperature-dependent breakdown of hydrogen peroxide-treated ZnO and TiO2 nanoparticle agglomerates

Metal oxide nanoparticles (MONPs) are used in a variety of applications including drug formulations, paint, sensors and biomedical devices due to their unique physicochemical properties. One of the major problems with their widespread implementation is their uncontrolled agglomeration. One approach to reduce agglomeration is to alter their surface chemistry with a proper functionality in an environmentally friendly way. In this study, the influence of hydrogen peroxide (H2O2) treatment on the dispersion of ZnO and TiO2 nanoparticle (NP) agglomerates as a function of temperature is studied. The H2O2 treatment of the MONPs increases the density of hydroxyl (–OH) groups on the NP surface, as verified with FTIR spectroscopy. The influence of heating on the dispersion of H2O2-treated ZnO and TiO2 NPs is investigated using dynamic light scattering. The untreated and H2O2-treated ZnO and TiO2 NP suspensions were heated from 30 °C to 90 °C at 5 °C intervals to monitor the breakdown of large aggregates into smaller aggregates and individual nanoparticles. It was shown that the combined effect of hydroxylation and heating enhances the dispersion of ZnO and TiO2 NPs in water

Growth of NBT template particles through topochemical microcrystal conversion and their structural characterization

In this study, uniform rectangular plate-like perovskite Na0.5Bi0.5TiO3 templates were obtained by molten salt synthesis method in three steps. Perovskite plate-like Na0.5Bi0.5TiO3 templates were synthesized from bismuth-layered Na0.5Bi4.5Ti4O15 by topochemical microcrystal conversion method at the third and final step. The most important point was that the plate-like morphology remained as desired, but templates were obtained in cubic perovskite structure. The Na2CO3 excess in the third step was the main controlling parameter. The crystalline orientation of the particles was investigated by electron backscatter diffraction analysis. The results indicated that the particles are single crystalline in nature. In the case of the 90mol% excess of the stoichiometric Na2CO3 ratio, the NBT platelets had highly regular, rectangular prismatic morphology and single-crystal nature. The structure was also confirmed through high-resolution transmission electron microscope and selected area electron diffraction analysis

Ultrafast Background-Free Ultrasound Imaging Using Blinking Nanoparticles

Localization-based ultrasound imaging methods that use microbubbles or nanodroplets offer high-resolution imaging with improved sensitivity and reduced background signal. However, these methods require long acquisition times (typically seconds to minutes), preventing their use for real-time imaging and, thus, limiting their clinical translational potential. Here, we present a new ultrafast localization method using blinking ultrasound-responsive nanoparticles (BNPs). When activated with high frame rate (1 kHz) plane wave ultrasound pulses with a mechanical index of 1.5, the BNPs incept growth of micrometer-sized bubbles, which in turn collapse and generate a blinking ultrasound signal. We showed that background-free ultrasound images could be obtained by localizing these blinking events using acquisition times as low as 11 ms. In addition, we demonstrated that BNPs enable in vivo background-free ultrasound imaging in mice. We envision that BNPs will facilitate the clinical translation of localization-based ultrasound imaging for more sensitive detection of cancer and other diseases

Ultrafast Background-Free Ultrasound Imaging Using Blinking Nanoparticles

Localization-based ultrasound imaging methods that use microbubbles or nanodroplets offer high-resolution imaging with improved sensitivity and reduced background signal. However, these methods require long acquisition times (typically seconds to minutes), preventing their use for real-time imaging and, thus, limiting their clinical translational potential. Here, we present a new ultrafast localization method using blinking ultrasound-responsive nanoparticles (BNPs). When activated with high frame rate (1 kHz) plane wave ultrasound pulses with a mechanical index of 1.5, the BNPs incept growth of micrometer-sized bubbles, which in turn collapse and generate a blinking ultrasound signal. We showed that background-free ultrasound images could be obtained by localizing these blinking events using acquisition times as low as 11 ms. In addition, we demonstrated that BNPs enable in vivo background-free ultrasound imaging in mice. We envision that BNPs will facilitate the clinical translation of localization-based ultrasound imaging for more sensitive detection of cancer and other diseases

Ultrafast Background-Free Ultrasound Imaging Using Blinking Nanoparticles

Localization-based ultrasound imaging methods that use microbubbles or nanodroplets offer high-resolution imaging with improved sensitivity and reduced background signal. However, these methods require long acquisition times (typically seconds to minutes), preventing their use for real-time imaging and, thus, limiting their clinical translational potential. Here, we present a new ultrafast localization method using blinking ultrasound-responsive nanoparticles (BNPs). When activated with high frame rate (1 kHz) plane wave ultrasound pulses with a mechanical index of 1.5, the BNPs incept growth of micrometer-sized bubbles, which in turn collapse and generate a blinking ultrasound signal. We showed that background-free ultrasound images could be obtained by localizing these blinking events using acquisition times as low as 11 ms. In addition, we demonstrated that BNPs enable in vivo background-free ultrasound imaging in mice. We envision that BNPs will facilitate the clinical translation of localization-based ultrasound imaging for more sensitive detection of cancer and other diseases

Ultrafast Background-Free Ultrasound Imaging Using Blinking Nanoparticles

Localization-based ultrasound imaging methods that use microbubbles or nanodroplets offer high-resolution imaging with improved sensitivity and reduced background signal. However, these methods require long acquisition times (typically seconds to minutes), preventing their use for real-time imaging and, thus, limiting their clinical translational potential. Here, we present a new ultrafast localization method using blinking ultrasound-responsive nanoparticles (BNPs). When activated with high frame rate (1 kHz) plane wave ultrasound pulses with a mechanical index of 1.5, the BNPs incept growth of micrometer-sized bubbles, which in turn collapse and generate a blinking ultrasound signal. We showed that background-free ultrasound images could be obtained by localizing these blinking events using acquisition times as low as 11 ms. In addition, we demonstrated that BNPs enable in vivo background-free ultrasound imaging in mice. We envision that BNPs will facilitate the clinical translation of localization-based ultrasound imaging for more sensitive detection of cancer and other diseases

Artificial Metalloproteins with Dinuclear Iron–Hydroxido Centers

Dinuclear iron centers with a bridging hydroxido or oxido ligand form active sites within a variety of metalloproteins. A key feature of these sites is the ability of the protein to control the structures around the Fe centers, which leads to entatic states that are essential for function. To simulate this controlled environment, artificial proteins have been engineered using biotin-streptavidin (Sav) technology in which Fe complexes from adjacent subunits can assemble to form [FeIII-(μ-OH)-FeIII] cores. The assembly process is promoted by the site-specific localization of the Fe complexes within a subunit through the designed mutation of a tyrosinate side chain to coordinate the Fe centers. An important outcome is that the Sav host can regulate the Fe···Fe separation, which is known to be important for function in natural metalloproteins. Spectroscopic and structural studies from X-ray diffraction methods revealed uncommonly long Fe···Fe separations that change by less than 0.3 Å upon the binding of additional bridging ligands. The structural constraints imposed by the protein host on the di-Fe cores are unique and create examples of active sites having entatic states within engineered artificial metalloproteins

2019 Turkish Hypertension Consensus Report

The Turkish Hypertension Consensus Report was prepared for the first time in 2015 to adapt the European and American international guidelines to our clinical practice and to create a practical report that could be a basic reference for all physicians dealing with hypertensive patients. This report, which was prepared by a committee with representation from 5 leading hypertension associations, has been accepted and is widely used. New clinical studies in hypertension literature and updated international guidelines since 2015 have demanded an update of the Turkish Hypertension Consensus Report as well. In this updated 2019 report, blood pressure levels were classified as Normal, Elevated, Stage 1, and Stage 2 hypertension. A new section was added for secondary hypertension. It was specified that drug treatment may be initiated with any 1 or a combination of 4 groups of drugs (diuretics, calcium channel blockers, angiotensin-converting enzyme [ACE] inhibitors, and angiotensin receptor blockers [ARBs]), except a combination of an ACE inhibitor and an ARB. It was emphasized that beta-blockers may be a first choice for hypertension treatment in diseases such as atrial fibrillation, heart failure, and coronary artery disease. The initial recommendation for hypertension treatment is a combination therapy in patients with a blood pressure level >= 150/90 mmHg. Target blood pressure values were redefined according to age and the presence of comorbidities. The hypertension treatment algorithm was renewed; it is proposed that drug therapy can also be initiated with a risk-based approach for the group with an elevated blood pressure (systolic blood pressure: 120-139 mmHg, diastolic blood pressure: 80-89 mmHg). The threshold clinic systolic blood pressure level was reduced from 160 mmHg to >= 150 mmHg for the initiation of drug therapy in individuals 80 years of age or more. The section on the treatment of special groups has now been expanded to include pregnancy and lactation. As in the previous report, in this update, practical recommendations for the most common cases seen in the clinic were the goal, rather than a comprehensive report that addresses all aspects of hypertension. This report has evidence-based recommendations for most patients; however, it should be kept in mind that there may be differences from 1 patient to another and that physicians should take an individualized approach according to a good clinical evaluation