scholarly journals Perchlorinated Triarylmethyl Radical 99% Enriched 13C at the Central Carbon as EPR Spin Probe Highly Sensitive to Molecular Tumbling

2021 ◽  
Author(s):  
Martin Poncelet ◽  
Justin L. Huffman ◽  
Gareth R. Eaton ◽  
Whylder Moore ◽  
Sandra Eaton ◽  
...  
Keyword(s):  
Spin Probe ◽  
High Sensitivity ◽  
Rotational Correlation Time ◽  
Spin Labels ◽  
Spectral Simulation ◽  
Epr Spectrum ◽  
Paramagnetic Resonance ◽  
Central Carbon

<p>Soluble stable radicals are used as spin probes and spin labels for <i>in vitro</i> and <i>in vivo</i> Electron Paramagnetic Resonance (EPR) spectroscopy and imaging applications. We report the synthesis and characterization of a perchlorinated triarylmethyl radical enriched 99% at the central carbon, <b><sup>13</sup>C<sub>1</sub>-PTMTC</b>. The anisotropy of the hyperfine splitting with the <sup>13</sup>C<sub>1</sub> (A<sub>x</sub>=26, A<sub>y</sub>=25, A<sub>z</sub>=199.5 MHz) and the g (g<sub>x</sub>=2.0015, g<sub>y</sub>=2.0015, g<sub>z</sub>=2.0040) are responsible for a strong effect of the radical tumbling rate on the EPR spectrum. The rotational correlation time can be determine by spectral simulation or via the linewidth after calibration. As spin probe <b><sup>13</sup>C<sub>1</sub>-PTMTC </b>can be used to measure media microviscosity with high sensitivity. Bound to a macromolecule as spin label, <b><sup>13</sup>C<sub>1</sub>-PTMTC </b>could be used to study local mobility and molecular interactions.</p>

scholarly journals Perchlorinated Triarylmethyl Radical 99% Enriched 13C at the Central Carbon as EPR Spin Probe Highly Sensitive to Molecular Tumbling

2021 ◽  
Author(s):  
Martin Poncelet ◽  
Justin L. Huffman ◽  
Gareth R. Eaton ◽  
Whylder Moore ◽  
Sandra Eaton ◽  
...  
Keyword(s):  
Spin Probe ◽  
High Sensitivity ◽  
Rotational Correlation Time ◽  
Spin Labels ◽  
Spectral Simulation ◽  
Epr Spectrum ◽  
Paramagnetic Resonance ◽  
Central Carbon

<p>Soluble stable radicals are used as spin probes and spin labels for <i>in vitro</i> and <i>in vivo</i> Electron Paramagnetic Resonance (EPR) spectroscopy and imaging applications. We report the synthesis and characterization of a perchlorinated triarylmethyl radical enriched 99% at the central carbon, <b><sup>13</sup>C<sub>1</sub>-PTMTC</b>. The anisotropy of the hyperfine splitting with the <sup>13</sup>C<sub>1</sub> (A<sub>x</sub>=26, A<sub>y</sub>=25, A<sub>z</sub>=199.5 MHz) and the g (g<sub>x</sub>=2.0015, g<sub>y</sub>=2.0015, g<sub>z</sub>=2.0040) are responsible for a strong effect of the radical tumbling rate on the EPR spectrum. The rotational correlation time can be determine by spectral simulation or via the linewidth after calibration. As spin probe <b><sup>13</sup>C<sub>1</sub>-PTMTC </b>can be used to measure media microviscosity with high sensitivity. Bound to a macromolecule as spin label, <b><sup>13</sup>C<sub>1</sub>-PTMTC </b>could be used to study local mobility and molecular interactions.</p>

scholarly journals Biological Applications of Electron Paramagnetic Resonance Viscometry Using a 13C-Labeled Trityl Spin Probe

Molecules ◽  
2021 ◽  
Vol 26 (9) ◽  
pp. 2781
Author(s):  
Murugesan Velayutham ◽  
Martin Poncelet ◽  
Timothy D. Eubank ◽  
Benoit Driesschaert ◽  
Valery V. Khramtsov
Keyword(s):  
Electron Paramagnetic Resonance ◽  
Spin Probe ◽  
Biological Fluids ◽  
Clinical Settings ◽  
Epr Spectrum ◽  
Paramagnetic Resonance ◽  
Field L ◽  
Living Tissues ◽  
Electron Paramagnetic

Alterations in viscosity of biological fluids and tissues play an important role in health and diseases. It has been demonstrated that the electron paramagnetic resonance (EPR) spectrum of a 13C-labeled trityl spin probe (13C-dFT) is highly sensitive to the local viscosity of its microenvironment. In the present study, we demonstrate that X-band (9.5 GHz) EPR viscometry using 13C-dFT provides a simple tool to accurately measure the microviscosity of human blood in microliter volumes obtained from healthy volunteers. An application of low-field L-band (1.2 GHz) EPR with a penetration depth of 1–2 cm allowed for microviscosity measurements using 13C-dFT in the living tissues from isolated organs and in vivo in anesthetized mice. In summary, this study demonstrates that EPR viscometry using a 13C-dFT probe can be used to noninvasively and rapidly measure the microviscosity of blood and interstitial fluids in living tissues and potentially to evaluate this biophysical marker of microenvironment under various physiological and pathological conditions in preclinical and clinical settings.

ДИНИТРОЗИЛЬНЫЕ КОМПЛЕКСЫ ЖЕЛЕЗА C ТИОЛСОДЕРЖАЩИМИ ЛИГАНДАМИ ПРЕДСТАВЛЕНЫ В ЖИВЫХ ОРГАНИЗМАХ В ОСНОВНОМ ИХ БИЯДЕРНОЙ ФОРМОЙ

2020 ◽  
Vol 65 (6) ◽  
pp. 1142-1153
Author(s):  
В.Д. Микоян ◽  
◽  
Е.Н. Бургова ◽  
Р.Р. Бородулин ◽  
А.Ф. Ванин ◽  
...  
Keyword(s):  
Electron Paramagnetic Resonance ◽  
Sodium Nitrite ◽  
Iron Complexes ◽  
Paramagnetic Resonance ◽  
First Case ◽  
Dinitrosyl Iron ◽  
Electron Paramagnetic ◽  
Dinitrosyl Complexes

The number of mononitrosyl iron complexes with diethyldithiocarbamate, formed in the liver of mice in vivo and in vitro after intraperitoneal injection of binuclear dinitrosyl iron complexes with N-acetyl-L-cysteine or glutathione, S-nitrosoglutathione, sodium nitrite or the vasodilating drug Isoket® was assessed by electron paramagnetic resonance (EPR). The number of the said complexes, in contrast to the complexes, formed after nitrite or Isoket administration, the level of which sharply increased after treatment of liver preparations with a strong reducing agent - dithionite, did not change in the presence of dithionite. It was concluded that, in the first case, EPR-detectable mononitrosyl iron complexes with diethyldithiocarbamate in the absence and presence of dithionite appeared as a result of the reaction of NO formed from nitrite with Fe2+-dieth- yldithiocarbamate and Fe3+-diethyldithiocarbamate complexes, respectively. In the second case, mononitrosyl iron complexes with diethyldithiocarbamate appeared as a result of the transition of iron-mononitosyl fragments from ready-made iron-dinitrosyl groups of binuclear dinitrosyl complexes, which is three to four times higher than the content of the mononuclear form of these complexes in the tissue...

scholarly journals Biological nanoscale fluorescent probes: From structure and performance to bioimaging

2020 ◽  
Vol 39 (1) ◽  
pp. 209-221
Author(s):  
Jiafeng Wan ◽  
Xiaoyuan Zhang ◽  
Kai Zhang ◽  
Zhiqiang Su
Keyword(s):  
Fluorescent Probes ◽  
Organic Dyes ◽  
Resonance Energy Transfer ◽  
Resonance Energy ◽  
High Sensitivity ◽  
Biological Imaging ◽  
Fluorescent Materials ◽  
And Performance

Abstract In recent years, nanomaterials have attracted lots of attention from researchers due to their unique properties. Nanometer fluorescent materials, such as organic dyes, semiconductor quantum dots (QDs), metal nano-clusters (MNCs), carbon dots (CDs), etc., are widely used in biological imaging due to their high sensitivity, short response time, and excellent accuracy. Nanometer fluorescent probes can not only perform in vitro imaging of organisms but also achieve in vivo imaging. This provides medical staff with great convenience in cancer treatment. Combined with contemporary medical methods, faster and more effective treatment of cancer is achievable. This article explains the response mechanism of three-nanometer fluorescent probes: the principle of induced electron transfer (PET), the principle of fluorescence resonance energy transfer (FRET), and the principle of intramolecular charge transfer (ICT), showing the semiconductor QDs, precious MNCs, and CDs. The excellent performance of the three kinds of nano fluorescent materials in biological imaging is highlighted, and the application of these three kinds of nano fluorescent probes in targeted biological imaging is also introduced. Nanometer fluorescent materials will show their significance in the field of biomedicine.

scholarly journals Bacteriophage-derived CHAP domain protein, P128, kills Staphylococcus cells by cleaving interpeptide cross-bridge of peptidoglycan

Microbiology ◽  
2014 ◽  
Vol 160 (10) ◽  
pp. 2157-2169 ◽  
Author(s):  
Sudarson Sundarrajan ◽  
Junjappa Raghupatil ◽  
Aradhana Vipra ◽  
Nagalakshmi Narasimhaswamy ◽  
Sanjeev Saravanan ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Mechanism Of Action ◽  
Catalytic Domain ◽  
Antibiotic Sensitivity ◽  
High Sensitivity ◽  
Common Mechanism ◽  
Cross Bridge ◽  
Sensitivity Pattern

P128 is an anti-staphylococcal protein consisting of the Staphylococcus aureus phage-K-derived tail-associated muralytic enzyme (TAME) catalytic domain (Lys16) fused with the cell-wall-binding SH3b domain of lysostaphin. In order to understand the mechanism of action and emergence of resistance to P128, we isolated mutants of Staphylococcus spp., including meticillin-resistant Staphylococcus aureus (MRSA), resistant to P128. In addition to P128, the mutants also showed resistance to Lys16, the catalytic domain of P128. The mutants showed loss of fitness as shown by reduced rate of growth in vitro. One of the mutants tested was found to show reduced virulence in animal models of S. aureus septicaemia suggesting loss of fitness in vivo as well. Analysis of the antibiotic sensitivity pattern showed that the mutants derived from MRSA strains had become sensitive to meticillin and other β-lactams. Interestingly, the mutant cells were resistant to the lytic action of phage K, although the phage was able to adsorb to these cells. Sequencing of the femA gene of three P128-resistant mutants showed either a truncation or deletion in femA, suggesting that improper cross-bridge formation in S. aureus could be causing resistance to P128. Using glutathione S-transferase (GST) fusion peptides as substrates it was found that both P128 and Lys16 were capable of cleaving a pentaglycine sequence, suggesting that P128 might be killing S. aureus by cleaving the pentaglycine cross-bridge of peptidoglycan. Moreover, peptides corresponding to the reported cross-bridge of Staphylococcus haemolyticus (GGSGG, AGSGG), which were not cleaved by lysostaphin, were cleaved efficiently by P128. This was also reflected in high sensitivity of S. haemolyticus to P128. This showed that in spite of sharing a common mechanism of action with lysostaphin, P128 has unique properties, which allow it to act on certain lysostaphin-resistant Staphylococcus strains.

scholarly journals Transitional B cells are the target of negative selection in the B cell compartment.

1995 ◽  
Vol 181 (6) ◽  
pp. 2129-2140 ◽  
Author(s):  
R Carsetti ◽  
G Köhler ◽  
M C Lamers
Keyword(s):  
B Cells ◽  
Negative Selection ◽  
Tolerance Induction ◽  
High Sensitivity ◽  
Target Population ◽  
Fas Antigen ◽  
Transitional B Cells ◽  
Immature B Cells

B lymphocytes recognize antigen through membrane-bound antigen-receptors, membrane IgM and IgD (mIgM and mIgD). Binding to foreign antigens initiates a cascade of biochemical events that lead to activation and differentiation. In contrast, binding to self-antigens leads to death or to inactivation. It is commonly believed that the B cells acquire the ability to discriminate between self and nonself in the early phases of development. We report here that immature B cells, which have just emerged from the mIgMneg, B220pos pool, are not deleted upon binding of self-antigen. In vivo, developing B cells become sensitive to tolerance induction in a relatively late window of differentiation, when they are in transition from the immature (HSAbright, B220dull) to the mature (HSAdull, B220bright) stage. In the transitional B cells, early markers of differentiation such as Pgp1 (CD44) and ThB reach the highest level of expression, while the expression of CD23 and mIgD, late markers of differentiation, and expression of class II MHC, progressively increases. Most of the transitional B cells, but only few of the mature and of the immature B cells, express the fas antigen, while mature B cells, but not immature and transitional B cells, express bcl-2 protein. mIgM is present in low amounts in immature B cells, reaches the highest level of expression in transitional B cells and is down-regulated in mature resting B cells, where it is coexpressed with mIgD. The high expression of mIgM, the presence of the fas antigen and the absence of bcl-2 protein is compatible with the high sensitivity of transitional B cells to negative selection. In vitro, immature B cells die rapidly by apoptosis after cross-linking of mIgM. This result, combined with the resistance of immature B cells to elimination in vivo, suggests that early in development the stroma cell microenvironment modulates signals transduced through mIgM. The functional and phenotypic division of IgMpos bone marrow B cells in three compartments not only allows to define the target population of physiological processes like negative selection, but will also be a helpful tool for an accurate description of possible developmental blocks in mutant mice.

Measurement of the Optical Properties of Muscle Tissue In Vivo

2000 ◽  
Author(s):  
P. L. Kopsombut ◽  
D. Willis ◽  
A. E. Schen ◽  
L. X. Xu ◽  
X. Xu
Keyword(s):  
Optical Properties ◽  
Transport Theory ◽  
Rapid Development ◽  
Ccd Camera ◽  
High Sensitivity ◽  
Biological Tissues ◽  
In Vivo Measurement ◽  
Living Tissues

Abstract Along with rapid development of diagnostic and therapeutic applications of lasers in medicine, optical properties of various biological tissues have been extensively studied [1]. Most of the studies were performed in vitro owing to the complexity involved in in vivo measurement. To date, it is well understood that living tissue is an absorbing and scattering heterogeneous medium because of its complex structures including blood network. The transport theory cannot be readily used due to the heterogeneity and the absence of the optical properties of living tissues [2]. In this research, we have developed a procedure for measuring the total attenuation coefficient (μ1) of the exteriorized rat 2-D spinotrapezius muscle in the wavelength ranged from 480–560 nm using the collimated light from a Nitrogen-pumped dye laser and a high-sensitivity CCD camera.

scholarly journals Nitric oxide interactions with cobalamins: biochemical and functional consequences

Blood ◽  
1996 ◽  
Vol 88 (5) ◽  
pp. 1857-1864 ◽  
Author(s):  
M Brouwer ◽  
W Chamulitrat ◽  
G Ferruzzi ◽  
DL Sauls ◽  
JB Weinberg
Keyword(s):  
Nitric Oxide ◽  
Electron Paramagnetic Resonance Spectroscopy ◽  
Resonance Spectroscopy ◽  
Paramagnetic Resonance ◽  
No Formation ◽  
Iron Nitrosyl ◽  
Wide Range ◽  
Functional Consequences

Abstract Nitric oxide (NO) is a paramagnetic gas that has been implicated in a wide range of biologic functions. The common pathway to evoke the functional response frequently involves the formation of an iron- nitrosyl complex in a target (heme) protein. In this study, we report on the interactions between NO and cobalt-containing vitamin B12 derivatives. Absorption spectroscopy showed that of the four Co(III) derivatives (cyanocobalamin [CN-Cbl], aquocobalamin [H2O-Cbl], adenosylcobalamin [Ado-Cbl], and methylcobalamin [MeCbl]), only the H2O- Cbl combined with NO. In addition, electron paramagnetic resonance spectroscopy of H2O-Cbl preparations showed the presence of a small amount of Cob-(II)alamin that was capable of combining with NO. The Co(III)-NO complex was very stable, but could transfer its NO moiety to hemoglobin (Hb). The transfer was accompanied by a reduction of the Co(III) to Co(II), indicating that NO+ (nitrosonium) was the leaving group. In accordance with this, the NO did not combine with the Hb Fe(II)-heme, but most likely with the Hb cysteine-thiolate. Similarly, the Co(III)-NO complex was capable of transferring its NO to glutathione. Ado-Cbl and Me-Cbl were susceptible to photolysis, but CN- Cbl and H2O-Cbl were not. The homolytic cleavage of the Co(III)-Ado or Co(III)-Me bond resulted in the reduction of the metal. When photolysis was performed in the presence of NO, formation of NO-Co(II) was observed. Co(II)-nitrosyl oxidized slowly to form Co(III)-nitrosyl. The capability of aquocobalamin to combine with NO had functional consequences. We found that nitrosylcobalamin had diminished ability to serve as a cofactor for the enzyme methionine synthase, and that aquocobalamin could quench NO-mediated inhibition of cell proliferation. Our in vitro studies therefore suggest that interactions between NO and cobalamins may have important consequences in vivo.

What can we Predict Before it's Implanted?

1985 ◽  
Vol 55 ◽  
Author(s):  
Donald F. Gibbons
Keyword(s):  
Biological Response ◽  
High Sensitivity ◽  
Biological Pathways ◽  
Surface Topology ◽  
Dystrophic Calcification ◽  
Vitro Assay ◽  
In Vivo Assays ◽  
Tissue Interface

ABSTRACTThe material factors which relate to the degradation and/or leaching of ions or molecules are described and the possible biological pathways which they may activate are described, i.e. cytotoxic, immune, tumor and nonspecific inflammatory response. Cytotoxicity is the only biological response which may be measured with high sensitivity by an in vitro assay prior to implantation. All other biological pathways require some degree of in vivo involvement. Three examples of biological response to material factors associated with devices which require evaluation by in vivo assays are discussed, namely: surface topology (texture), mechanically induced factors at the device/tissue interface caused by differences in compliance, and dystrophic calcification in connective tissue and vascular devices.

scholarly journals A novel decatenation assay for DNA topoisomerases using a singly-linked catenated substrate

BioTechniques ◽  
2020 ◽  
Vol 69 (5) ◽  
pp. 356-362
Author(s):  
Nidda F Waraich ◽  
Shruti Jain ◽  
Sean D Colloms ◽  
William Marshall Stark ◽  
Nicolas P Burton ◽  
...  
Keyword(s):  
High Sensitivity ◽  
Enzyme Digestion ◽  
Restriction Enzyme Digestion ◽  
Kinetoplast Dna ◽  
Dna Topoisomerases ◽  
In Vitro Drug Screening ◽  
Drug Inhibition ◽  
Dna Substrate

Decatenation is a crucial in vivo reaction of DNA topoisomerases in DNA replication and is frequently used in in vitro drug screening. Usually this reaction is monitored using kinetoplast DNA as a substrate, although this assay has several limitations. Here we have engineered a substrate for Tn 3 resolvase that generates a singly-linked catenane that can readily be purified from the DNA substrate after restriction enzyme digestion and centrifugation. We show that this catenated substrate can be used with high sensitivity in topoisomerase assays and drug-inhibition assays.

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