scholarly journals Fluorescent magnesium nanocomplex in a protein scaffold for cell nuclei imaging applications

RSC Advances ◽  
2015 ◽  
Vol 5 (114) ◽  
pp. 94236-94240 ◽  
Author(s):  
Alok Pandya ◽  
Apritam Tripathi ◽  
Rahul Purohit ◽  
Sanjay Singh ◽  
Manjula I. Nandasiri ◽  
...  
Keyword(s):  
Protein Complex ◽  
High Specificity ◽  
Cell Nuclei ◽  
Protein Scaffold ◽  
Photo Stability ◽  
First Time ◽  
Magnesium Nanoparticles

For the first time, we report a facile strategy for the synthesis of ultra-fine blue-green emitting fluorescent magnesium nanoparticles–protein complex (MgNC) which shows excellent photo stability and also stain cell nuclei with high specificity.

scholarly journals Immunoglobulins anti-endonuclease 32 kDa from Cucurbita pepo var patissonina affect process of DNA synthesis.

1995 ◽  
Vol 42 (2) ◽  
pp. 177-182
Author(s):  
R Rzepecki ◽  
J Szopa
Keyword(s):  
Dna Synthesis ◽  
Nuclear Matrix ◽  
Protein Complex ◽  
Cucurbita Pepo ◽  
Dna Fragments ◽  
Cell Nuclei ◽  
Structural Organisation ◽  
Low Concentrations

Immunoglobulins anti-endonuclease 32 kDa inhibit DNA synthesis. We observed that low concentrations of IgGs (about 50 micrograms IgG per 1 x 10(6) cell nuclei) temporary inhibit DNA synthesis. This inhibition concerns only the synthesis of DNA bound to the nuclear matrix (associated with isolated nuclear matrix). Preincubation of cell nuclei of White bush with IgG generates longer DNA fragments than in controls. Involvement of the 32 kDa endonuclease or an endonuclease-65 kDa protein complex from the nuclear matrix in replication or structural organisation of replication is considered.

Identification of chromosomal actin in insect cells: Structural and functional significance

1994 ◽  
Vol 52 ◽  
pp. 16-17
Author(s):  
Ivo Sauman
Keyword(s):  
Insect Cells ◽  
Polytene Chromosomes ◽  
Follicle Cell ◽  
Nuclear Actin ◽  
Cell Nuclei ◽  
Antibody Staining ◽  
Eukaryotic Nucleus ◽  
Biochemical Studies ◽  
First Time ◽  
Endonuclease Digestion

The non-histone proteins associated with eukaryotic nuclear chromatin are incompletely characterized and their function is poorly understood. Thirty years ago, the presence of actin in the eukaryotic nucleus was reported for first time. Since then, several biochemical studies have identified actin and myosin as significant constituents of isolated nuclear matrix from a variety of cells. The studies cited above and others make a strong case for presence of actin in nuclei, but do not implicate actin as a component of eukaryotic chromosomes.Our examination of cells associated with developing ovarian follicles of lepidopterans confirmed that under routine immunocytochemical conditions, no actin can be detected with anti-actin antibodies in the follicle cell nuclei (Fig. 1a) . However, Fig. 1b demonstrates that endonuclease pretreatment of the same preparation to remove DNA followed by anti-actin antibody staining uncovers the presence of nuclear actin. Moreover, by employing squash preparations of Drosophila salivary glands and the same endonuclease digestion, it is clear that the nuclear actin is directly associated with the polytene chromosomes (Fig. 2a,b).

scholarly journals Diverse high-torque bacterial flagellar motors assemble wider stator rings using a conserved protein scaffold

2016 ◽  
Vol 113 (13) ◽  
pp. E1917-E1926 ◽  
Author(s):  
Morgan Beeby ◽  
Deborah A. Ribardo ◽  
Caitlin A. Brennan ◽  
Edward G. Ruby ◽  
Grant J. Jensen ◽  
...  
Keyword(s):  
Protein Scaffold ◽  
Related Family ◽  
Flagellar Motors ◽  
Torque Generation ◽  
High Torque ◽  
Protein Components ◽  
Subtomogram Averaging ◽  
First Time ◽  
Sequential Assembly

Although it is known that diverse bacterial flagellar motors produce different torques, the mechanism underlying torque variation is unknown. To understand this difference better, we combined genetic analyses with electron cryo-tomography subtomogram averaging to determine in situ structures of flagellar motors that produce different torques, fromCampylobacterandVibriospecies. For the first time, to our knowledge, our results unambiguously locate the torque-generating stator complexes and show that diverse high-torque motors use variants of an ancestrally related family of structures to scaffold incorporation of additional stator complexes at wider radii from the axial driveshaft than in the model enteric motor. We identify the protein components of these additional scaffold structures and elucidate their sequential assembly, demonstrating that they are required for stator-complex incorporation. These proteins are widespread, suggesting that different bacteria have tailored torques to specific environments by scaffolding alternative stator placement and number. Our results quantitatively account for different motor torques, complete the assignment of the locations of the major flagellar components, and provide crucial constraints for understanding mechanisms of torque generation and the evolution of multiprotein complexes.

THIN PREPARATIONS OF HEMOPOIETIC TISSUES

1961 ◽  
Vol 39 (3) ◽  
pp. 367-372 ◽  
Author(s):  
Vibeke E. Engelbert
Keyword(s):  
Toluidine Blue ◽  
Blue Staining ◽  
Cell Nuclei ◽  
Millipore Filter ◽  
Toluidine Blue Staining ◽  
Glass Slides ◽  
Cell Layers ◽  
First Time ◽  
Millipore Filters

Previous papers dealing with behaviors and activities of blood cell nuclei have been recording mainly results with the imprint method and observations of blood cells cultured in vitro. The present paper demonstrates that the Feulgen "gentle squash" method as well as digestion with N HCl followed by toluidine blue staining at pH 4 reveals the same nuclear behaviors as the imprint method. Imprinting on millipore filters with subsequent fixation in Zenker's acetic solution followed by the Feulgen or toluidine blue staining method (the latter with or without previous N HCl digestion) is reported for the first time. The millipore imprint preserves a thicker layer of cells than imprinting on glass slides. Also cell stages which may not easily stick to glass seem to adhere well to the millipore filter. The filter imprint may in places present as many cell layers as a thin slice cut on the microtome, but without the disadvantage of having large or elongated cell stages cut into pieces by the microtome knife.

scholarly journals Objective and Subjective Evaluation of Online Error Correction during P300-Based Spelling

2012 ◽  
Vol 2012 ◽  
pp. 1-13 ◽  
Author(s):  
Perrin Margaux ◽  
Maby Emmanuel ◽  
Daligault Sébastien ◽  
Bertrand Olivier ◽  
Mattout Jérémie
Keyword(s):  
Error Correction ◽  
Error Detection ◽  
Subjective Evaluation ◽  
High Specificity ◽  
Automatic Correction ◽  
Bit Rate ◽  
Computer Interfaces ◽  
Probabilistic Classifier ◽  
Online Error Detection ◽  
First Time

Error potentials (ErrP) are alterations of EEG traces following the subject’s perception of erroneous feedbacks. They provide a way to recognize misinterpreted commands in brain-computer interfaces (BCI). However, this has been evaluated online in only a couple of studies and mostly with very few subjects. In this study, we implemented a P300-based BCI, including not only online error detection but also, for the first time, automatic correction. We evaluated it in 16 healthy volunteers. Whenever an error was detected, a new decision was made based on the second best guess of a probabilistic classifier. At the group level, correction did neither improve nor deteriorate spelling accuracy. However, automatic correction yielded a higher bit rate than a respelling strategy. Furthermore, the fine examination of interindividual differences in the efficiency of error correction and spelling clearly distinguished between two groups who differed according to individual specificity in ErrP detection. The high specificity group had larger evoked responses and made fewer errors which were corrected more efficiently, yielding a 4% improvement in spelling accuracy and a higher bit rate. Altogether, our results suggest that the more the subject is engaged into the task, the more useful and well accepted the automatic error correction.

scholarly journals Biomimetic Sensitive Elements for 2,4,6-Trinitrotoluene Tested on Multi-Layered Sensors

Coatings ◽  
2020 ◽  
Vol 10 (3) ◽  
pp. 273
Author(s):  
Ana Mihaela GAVRILA ◽  
Tanta Verona IORDACHE ◽  
Carmen LAZAU ◽  
Traian ROTARIU ◽  
Ileana CERNICA ◽  
...  
Keyword(s):  
Technological Progress ◽  
Chemical Sensor ◽  
Response Times ◽  
High Specificity ◽  
Imprinted Polymer ◽  
Vapor State ◽  
Assembly Mechanism ◽  
Low Sensitivity ◽  
First Time ◽  
Tnt Detection

In spite of technological progress, most of the current techniques for 2,4,6-trinitrotoluene (TNT) detection are time consuming due to laborious sensor preparation. Thereby, the aim of this work was to enlarge the knowledge for preparing sensitive elements for TNT with the aid of molecular imprinting; a known technique used to deliver biomimetic materials. The study first depicts the auto-assembly mechanism of (TNT) with functional diamino-silanes (i.e., N-(2-aminoethyl)-3-aminopropyl methyl dimethoxysilane), via “double” Meisenheimer complexes. This mechanism is being described herein for the first time and applied further to obtain molecularly imprinted polymer (MIP) films for TNT recognition. For testing the potential application of films as chemical sensor elements, typical rebinding assays of TNT in a liquid state and the rebinding of TNT in a vapor state, using multilayered sensor chips composed of quartz-chromium (Cr)-gold (Au)-titanium oxide (TiO2), were employed. Batch rebinding experiments have shown that thinner films were more efficient on retaining TNT molecules in the first five min, with a specificity of about 1.90. The quartz-Cr-Au-TiO2-MIP capacitive sensors, tested in vapor state, registered short response times (less than 25 s), low sensitivity to humidity and high specificity for TNT.

Rational design of a lipid-droplet-polarity based fluorescent probe for potential cancer diagnosis

2018 ◽  
Vol 54 (85) ◽  
pp. 12093-12096 ◽  
Author(s):  
Junling Yin ◽  
Min Peng ◽  
Yanyan Ma ◽  
Rui Guo ◽  
Weiying Lin
Keyword(s):  
Fluorescent Probe ◽  
Lipid Droplet ◽  
Cancer Diagnosis ◽  
Rational Design ◽  
High Specificity ◽  
Diagnosis Of Cancer ◽  
First Time ◽  
Potential Cancer

We have rationally designed a robust fluorescent probe CTPA for potential cancer diagnosis by monitoring LD numbers and polarity variation. With the outstanding solvatochromism and high specificity for LDs of CTPA, the diagnosis of cancer can be achieved not only at the cellular levels but also in organs and living mice for the first time.

scholarly journals Atroposelective antibodies as a designed protein scaffold for artificial metalloenzymes

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Takuma Adachi ◽  
Akira Harada ◽  
Hiroyasu Yamaguchi
Keyword(s):  
Bond Formation ◽  
Metal Catalysts ◽  
Alkylation Reaction ◽  
Protein Scaffold ◽  
Asymmetric Catalysts ◽  
Reaction Proceeds ◽  
Cu Catalyst ◽  
Artificial Metalloenzymes ◽  
First Time ◽  
Protein Environment

Abstract Design and engineering of protein scaffolds are crucial to create artificial metalloenzymes. Herein we report the first example of C-C bond formation catalyzed by artificial metalloenzymes, which consist of monoclonal antibodies (mAbs) and C2 symmetric metal catalysts. Prepared as a tailored protein scaffold for a binaphthyl derivative (BN), mAbs bind metal catalysts bearing a 1,1′-bi-isoquinoline (BIQ) ligand to yield artificial metalloenzymes. These artificial metalloenzymes catalyze the Friedel-Crafts alkylation reaction. In the presence of mAb R44E1, the reaction proceeds with 88% ee. The reaction catalyzed by Cu-catalyst incorporated into the binding site of mAb R44E1 is found to show excellent enantioselectivity with 99% ee. The protein environment also enables the use of BIQ-based catalysts as asymmetric catalysts for the first time.

scholarly journals Development of High-Specificity Fluorescent Probes to Enable Cannabinoid Type 2 Receptor Studies in Living Cells

2020 ◽  
Author(s):  
Roman Sarott ◽  
Matthias Westphal ◽  
Patrick Pfaff ◽  
Claudia Korn ◽  
David Sykes ◽  
...  
Keyword(s):  
Fluorescent Probes ◽  
Modular Design ◽  
Inflammatory Diseases ◽  
High Specificity ◽  
Cell Types ◽  
Living Cells ◽  
Binding Assays ◽  
Mouse Splenocytes ◽  
First Time

Pharmacological modulation of cannabinoid type 2 receptor (CB<sub>2</sub>R) holds promise for the treatment of numerous conditions, including inflammatory diseases, autoimmune disorders, pain, and cancer. Despite the significance of this receptor, researchers lack reliable tools to address questions concerning the expression and complex mechanism of CB<sub>2</sub>R signaling, especially in cell-type and tissue-dependent context. Herein, we report for the first time a versatile ligand platform for the modular design of a collection of highly specific CB<sub>2</sub>R fluorescent probes, used successfully across applications, species and cell types. These include flow cytometry of endogenously expressing cells, real-time confocal microscopy of mouse splenocytes and human macrophages, as well as FRET-based kinetic and equilibrium binding assays. High CB<sub>2</sub>R specificity was demonstrated by competition experiments in living cells expressing CB<sub>2</sub>R at native levels. The probes were effectively applied to FACS analysis of microglial cells derived from a mouse model relevant to Alzheimer’s disease and to the detection of CB<sub>2</sub>R in human breast cancer cells.

Single-base mismatch discrimination by T7 exonuclease with target cyclic amplification detection

2015 ◽  
Vol 51 (14) ◽  
pp. 2954-2956 ◽  
Author(s):  
Zhen-Kun Wu ◽  
Dian-Ming Zhou ◽  
Zhan Wu ◽  
Xia Chu ◽  
Ru-Qin Yu ◽  
...  
Keyword(s):  
High Specificity ◽  
Single Base ◽  
Single Base Mismatch ◽  
Base Mismatch ◽  
Mismatch Discrimination ◽  
First Time ◽  
T7 Exonuclease

T7 exonuclease is reported for the first time to have high specificity in discriminating single-base mismatch.

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