Gentaur Pub

#34545108   2021/09/20 To Up

Inhaled sphingosine has no adverse side effects in isolated ventilated and perfused pig lungs.

Ex-vivo lung perfusion (EVLP) systems like XVIVO are more and more common in the setting of lung transplantation, since marginal donor-lungs can easily be subjected to a performance test or be treated with corticosteroids or antibiotics in high dose regimes. Donor lungs are frequently positive in bronchoalveolar lavage (BAL) bacterial cultures (46-89%) which leads to a donor-to-recipient transmission and after a higher risk of lung infection with reduced posttransplant outcome. We have previously shown that sphingosine very efficiently kills a variety of pathogens, including Pseudomonas aeruginosa, Staphylococcus aureus and epidermidis, Escherichia coli or Haemophilus influenzae. Thus, sphingosine could be a new treatment option with broadspectrum antiinfective potential, which may improve outcome after lung transplantation when administered prior to lung re-implantation. Here, we tested whether sphingosine has any adverse effects in the respiratory tract when applied into isolated ventilated and perfused lungs. A 4-h EVLP run using minipig lungs was performed. Functional parameters as well as perfusate measurements where obtained. Biopsies were obtained 30 min and 150 min after inhalation of sphingosine. Tissue samples were fixed in paraformaldehyde, embedded in paraffin and sectioned. Hemalaun, TUNEL as well as stainings with Cy3-coupled anti-sphingosine or anti-ceramide antibodies were implemented. We demonstrate that tube-inhalation of sphingosine into ex-vivo perfused and ventilated minipig lungs results in increased levels of sphingosine in the luminal membrane of bronchi and the trachea without morphological side effects up to very high doses of sphingosine. Sphingosine also did not affect functional lung performance. In summary, the inhalation of sphingosine results in an increase of sphingosine concentrations in the luminal plasma membrane of tracheal and bronchial epithelial cells. The inhalation has no local side effects in ex-vivo perfused and ventilated minipig lungs.
Henning Carstens, Katharina Kalka, Rabea Verhaegh, Fabian Schumacher, Matthias Soddemann, Barbara Wilker, Simone Keitsch, Carolin Sehl, Burkhard Kleuser, Thorsten Wahlers, Gerald Reiner, Achim Koch, Ursula Rauen, Erich Gulbins, Markus Kamler

1969 related Products with: Inhaled sphingosine has no adverse side effects in isolated ventilated and perfused pig lungs.

50 ul

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#34517663   2021/08/14 To Up

Combination of bidirectional strand displacement amplification with single-molecule detection for multiplexed DNA glycosylases assay.

DNA glycosylases can initiate base excision repair pathway to repair endogenous DNA base damages for the maintenance of genome stability. Multiple DNA glycosylases exhibit abnormal in various diseases, and the simultaneous measurement of different DNA glycosylases is critical to clinical diagnosis and drug discovery. Herein, we take advantage of single-molecule detection and bidirectional strand displacement amplification (SDA) to simultaneously detect uracil DNA glycolase (UDG) and human alkyladenine DNA glycosylase (hAAG). We design a partial double-stranded DNA (dsDNA) substrate modified with specific recognition sites of UDG and hAAG. The dsDNA substrate is labeled with BHQ1 and BHQ2 at the 5'-ends and then hybridizes with the Cy3/Cy5-labeled reporter probes to obtain the BHQ1/Cy3 and BHQ2/Cy5 base pairs, resulting in the quenching of Cy3/Cy5 fluorescence by BHQ1/BHQ2 via fluorescence resonance energy transfer (FRET). When UDG and hAAG are present, they can induce the base excision repair reaction and subsequently initiate the bidirectional SDA amplification process, releasing the Cy5/Cy3-labeled reporter probes from the dsDNA substrate and consequently the recovery of Cy5 and Cy3 fluorescence, which can be measured by single-molecule detection, with Cy5 indicating UDG and Cy3 indicating hAAG. This method possesses high sensitivity and good selectivity with the capability of quantifying multiple DNA glycosylases at the single-cell level. Furthermore, it can be used to simultaneously screen DNA glycosylase inhibitors and determine enzyme kinetic parameters, with the potential of sensing various DNA/RNA enzymes by simple changing the recognition sites of DNA substrates.
Yan Zhang, Jinping Hu, Xiao-Yun Yang, Chun-Yang Zhang

1290 related Products with: Combination of bidirectional strand displacement amplification with single-molecule detection for multiplexed DNA glycosylases assay.

100tests200 ug100 assays1,000 tests50 assays100tests100tests100tests100Tests2x96 well plate

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#34496977   2021/09/08 To Up

Aflibercept clearance through the drainage system in a rat model.

As intravitreal anti-VEGF injections became the mainstay of treatment for many retinal diseases, the cause of a secondary sustained elevated intraocular pressure is still unclear. The aim of our study was to study the clearance of Aflibercept from the anterior chamber angle, in a rat model, to test if an aggregation exists.
Yariv Keshet, Orly Gal-Or, Michal Schaap Fogler, Karin Mimouni, Meydan Ben Ishai, Dov Weinberger, Assaf Dotan

1902 related Products with: Aflibercept clearance through the drainage system in a rat model.

100 UG1

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#34492515   2021/08/31 To Up

A turn-on fluorescence sensor for rapid sensing of ATP based on luminescence resonance energy transfer between upconversion nanoparticles and Cy3 in vivo or vitro.

Adenosine triphosphate (ATP) is an energy molecule of significant importance, and, the monitoring of ATP in living cells is considerable for the clinical diagnosis of many related diseases, including cancer. Upconversion nanoparticles (UCNPs) have recently been attracting widespread interest in biomedical applications due to their chemical and thermal stability, high sensitivity, good biocompatibility, and excellent tissue penetration. Herein, a Cy3-aptamer-cDNA- UCNPs nanosensor was synthesized, based on the luminescence resonance energy transfer (LRET) between UCNPs and Cy3 for monitoring ATP in living cells. It showed a selective sensing ability for ATP levels by changes of fluorescence intensity of UNCPs at 536 nm. The investigated biosensor showed a precise, efficient detection with sufficient selectivity which was achieved through the optimization of conditions. In the range of 1-1000 μM, the ATP-induced changes of the fluorescence intensity were linearly proportional to the ATP concentrations. Furthermore, the cytotoxicity assay revealed that the UCNPs sensor exhibited favorable biocompatibility, implicating the use of UCNPs in vivo imaging. This study highlights the potential of using a combination of UCNPs and ATP-binding aptamer to design an ATP-activatable probe for fluorescence-mediated imaging in living cells. These results implied that the nanosensor can be applicable for the monitoring of intracellular ATP by fluorescence imaging and the quantitative analysis of biological liquids.
Jing Xu, Huanhuan Li, Selva Sharma Arumugam, Yawen Rong, Pingyue Wang, Quansheng Chen

1720 related Products with: A turn-on fluorescence sensor for rapid sensing of ATP based on luminescence resonance energy transfer between upconversion nanoparticles and Cy3 in vivo or vitro.

100ug1 kit100 plates100 μg

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#34487317   2021/09/06 To Up

Zika Virus NS1 Suppresses VE-Cadherin and Claudin-5 via hsa-miR-101-3p in Human Brain Microvascular Endothelial Cells.

Zika virus (ZIKV) is a neurotropic virus that causes microcephaly in newborns and Guillain-Barré syndrome (GBS) in adults. ZIKV is known to transmigrate through the blood-brain barrier (BBB) by utilizing different strategies. NS1 is a conserved flavivirus protein, which is secreted extracellularly. ZIKV-NS1 has been shown to target adherens junctions (AJs) and tight junctions (TJs) to disrupt the endothelial barrier integrity. The microRNAs are short non-coding RNAs, which post-transcriptionally regulate the gene expression by binding to 3' UTR of the target gene. In the present study, we studied the ZIKV-NS1-mediated effect through hsa-miR-101-3p on the junctional barrier integrity in human brain microvascular endothelial cells. We exposed hBMVECs and hCMEC/D3 cells with ZIKV-NS1 at different time points (12 h and 24 h) with the doses 500 ng/mL and 1000 ng/mL. The change in the expression of VE-cadherin and claudin-5 was quantified using immunoblotting. The expression of the hsa-miR-101-3p was quantified using qRT-PCR. To prove the targeting of hsa-miR-101-3p to VE-cadherin, we transfected hsa-miR-101-3p mimic, scramble, hsa-miR-101-3p inhibitor, and Cy3 in the ZIKV-NS1-exposed hCMEC/D3 cells. The distribution and expression of the VE-cadherin and claudin-5 were observed using immunofluorescence and immunoblotting. The ZIKV-NS1 compromises the endothelial barrier integrity by disrupting the VE-cadherin and claudin-5 protein expression via hsa-miR-101-3p. The findings of this study suggest that ZIKV-NS1 dysregulates the adherens junction and tight junction proteins through hsa-miR-101-3p, which compromises the barrier integrity of human brain microvascular endothelial cells.
Utkarsh Bhardwaj, Sunit K Singh

2355 related Products with: Zika Virus NS1 Suppresses VE-Cadherin and Claudin-5 via hsa-miR-101-3p in Human Brain Microvascular Endothelial Cells.

1.00 flask1.00 flask1.00 flask1.00 flask1.00 flask1.00 flask1.00 flask1.00 flask1.00 flask1.00 flask1.00 flask1.00 flask

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#34453919   2021/08/25 To Up

Comprehensive analysis of protein-expression changes specific to immunoglobulin G4-related disease.

Immunoglobulin 4 (IgG4)-related disease (IgG4-RD) is a lymphoproliferative disorder characterized by elevated serum IgG4 levels and tissue infiltration of IgG4-positive plasma cells. We analyzed the serum proteins, whose levels varied based on the disease state and treatment.
Takafumi Kawanami, Haruka Kawanami-Iwao, Takanobu Takata, Yasuhito Ishigaki, Naohisa Tomosugi, Tsutomu Takegami, Hiroto Yanagisawa, Shino Fujimoto, Tomoyuki Sakai, Yoshimasa Fujita, Kazunori Yamada, Shuichi Mizuta, Hiroshi Kawabata, Toshihiro Fukushima, Yuko Hirose, Yasufumi Masaki

2360 related Products with: Comprehensive analysis of protein-expression changes specific to immunoglobulin G4-related disease.

100 1 mg1 module1 module96T100 1 module2 Pieces/Box1 module1 module1 module1 module

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#34450954   2021/08/17 To Up

Development of Lectin Modified Fluorescent Magnetic Particles for Highly Sensitive Detection of Glycoconjugates.

I conducted this study to develop an improved method for glycome detection using fluorescent magnetic beads, whose surfaces were modified using lectins, for the highly sensitive detection of saccharides or glycoproteins via fluorescence quenching using a novel fluorescence emitter and quencher pair. The emitter (Cy3 fluorophore) was incorporated into magnetic beads, and a fluorescence quencher (cyanopyranyl group) was bound to glycomes via covalent bonding. The fluorescence intensities of fluorescent magnetic beads containing lectins decreased specifically in the presence of glycomes, which was a result of fluorescence quenching from Cy3 to cyanopyranyl groups due to the formation of a stable complex between lectins and glycome. Fluorescence intensities were plotted as a function of glycoprotein concentration, and good linear relationships were observed. This method enabled the fluorescent reading-out of a series of lectin-glycome interactions on the basis of recognition selectivity and affinity of immobilized lectins without tedious washing processes. Moreover, a simple profiling process was performed using this assay for diverse glycoconjugates, which not only included simple saccharides but also glycoproteins and glycome in cell lysates. These results clearly indicate that the combination of magnetic beads with the novel emitter-quencher pair enabled the highly sensitive detection of lectin-glycome interactions.
Yoshio Suzuki

1503 related Products with: Development of Lectin Modified Fluorescent Magnetic Particles for Highly Sensitive Detection of Glycoconjugates.

96 tests1x96 well plate96 wells 5 G

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#34432445   2021/08/25 To Up

Mechanism of Cyanine5 to Cyanine3 Photoconversion and Its Application for High-Density Single-Particle Tracking in a Living Cell.

Cyanine (Cy) dyes are among the most useful organic fluorophores that have found a wide range of applications in single-molecule and super-resolution imaging as well as in other biophysical studies. However, recent observations that blueshifted derivatives of Cy dyes are formed via photoconversion have raised concerns as to the potential artifacts in multicolor imaging. Here, we report the mechanism for the photoconversion of Cy5 to Cy3 that occurs upon photoexcitation during fluorescent imaging. Our studies show that the formal CH excision from Cy5 occurs mainly through an intermolecular pathway involving a combination of bond cleavage and reconstitution while unambiguously confirming the identity of the fluorescent photoproduct of Cy5 to be Cy3 using various spectroscopic tools. The carbonyl products generated from singlet oxygen-mediated photooxidation of Cy5 undergo a sequence of carbon-carbon bond-breaking and -forming events to bring about the novel dye-to-dye transformation. We also show that the deletion of a two-methine unit from the polymethine chain, which results in the formation of blueshifted products, commonly occurs in other cyanine dyes, such as Alexa Fluor 647 (AF647) and Cyanine5.5. The formation of a blueshifted congener dye can obscure the multicolor fluorescence imaging, leading to misinterpretation of the data. We demonstrate that the potentially deleterious photoconversion, however, can be exploited to develop a new photoactivation method for high-density single-particle tracking in a living cell without using UV illumination and cell-toxic additives.
Yoonjung Cho, Hyeong Jeon An, Taehoon Kim, Chulbom Lee, Nam Ki Lee

1228 related Products with: Mechanism of Cyanine5 to Cyanine3 Photoconversion and Its Application for High-Density Single-Particle Tracking in a Living Cell.

1 kit100ug Lyophilized

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