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localization microscopy

Monica Terracciano, Michela Napolitano, Luca De Stefano, Anna Chiara De Luca, Ilaria Rea
Diatomite is a fossil material made of amorphous porous silica. In this work, polyethylene glycol (PEG)-modified diatomite NPs (PEG-DNPs) are decorated with gold NPs (AuNPs) by one-pot liquid-phase synthesis. Nanocomplexes (PEG-DNPs@AuNPs), with an average size of about 450 nm, are characterized by dynamic light scattering, electron microscopy, nitrogen adsorption/desorption analysis, UV-VIS and photoluminescence spectroscopies. Preliminary studies on the use of the nanocomplex in nanomedicine are also presented...
March 19, 2018: Nanotechnology
Sreejesh Sreedharan, Alessandro Sinopoli, Paul J Jarman, Darren Robinson, Christopher Clemmet, Paul A Scattergood, Craig R Rice, Carl G W Smythe, James A Thomas, Paul I P Elliott
Two new biscyclometalated complexes [Ir(ptzR)2(dppz)]+ (dppz = dipyridophenazene; ptzRH = 4-phenyl-1-benzyl-1,2,3-triazole (1+) and 4-phenyl-1-propyl-1,2,3-triazole (2+)) have been prepared. The hexafluorophosphate salts of these complexes have been fully characterized and, in one case, the X-ray structure of a nitrate salt was obtained. The DNA binding properties of the chloride salts of the complexes were investigated, as well as their cellular uptake by A2780 and MCF7 cell lines. Both complexes display an increase in the intensity of phosphorescence upon titration with duplex DNA, indicating the intercalation of the dppz ligand and, given that they are monocations, the complexes exhibit appreciable DNA binding affinity...
March 19, 2018: Dalton Transactions: An International Journal of Inorganic Chemistry
Amandine Bastide, Jonathan W Yewdell, Alexandre David
While isotopic labeling of amino acids remains the reference method in the field for quantifying translation rate, it does not provide any information on spatial localization of translation sites. The rationale behind developing the ribopuromycylation method (RPM) was primarily to map translation sites at the sub-cellular level while avoiding detection of newly synthesized proteins released from ribosomes. RPM visualizes actively translating ribosomes in cells via standard immunofluorescence microscopy in fixed and permeabilized cells using a puromycin-specific monoclonal antibody to detect puromycylated nascent chains trapped on ribosomes treated with a chain elongation inhibitor...
January 5, 2018: Bio-protocol
Roland Thuenauer, Simon Nicklaus, Marco Frensch, Kevin Troendle, Josef Madl, Winfried Römer
A key factor determining the fate of individual cells within an epithelium is the unique microenvironment that surrounds each cell. It regulates location-dependent differentiation into specific cellular sub-types, but, on the other hand, a disturbed microenvironment can promote malignant transformation of epithelial cells leading to cancer formation. Here, we present a tool based on a microfluidic biochip that enables novel research approaches by providing a means to control the basolateral microenvironment of a confined number of neighbouring cells within an epithelial monolayer...
February 19, 2018: RSC Advances
Joshua T Byers, Xin Qing, Christopher Lo, Samuel W French, Ping Ji
Amyloidosis is a disorder characterized by the deposition of insoluble abnormal proteins in the extracellular space. It may occur as a localized lesion or as a systemic disease involving multiple organs and systems. Localized conjunctival amyloidosis is rare and is less frequently associated with systemic involvement. Although amyloidosis itself is a benign lesion involvement of multiple organs and systems is associated with poor prognosis. Diagnosis of amyloidosis is made on biopsy specimens with Congo red staining for the appearance of apple-green birefringence under polarized light microscopy...
March 15, 2018: Experimental and Molecular Pathology
Kartikeya Tiwari, Vikash Kumar Dubey
Asparaginase, a pivotal enzyme of the aspartate metabolic pathway is present as two variants in Leishmania donovani, causative agent of visceral leishmaniasis (VL). Our microscopy based analysis revealed that the green fluorescent protein tagged asparaginase variants were localized in the cytosol of the parasite. The finding was also supported by the in silico analysis of the L. donovani asparaginase variants. This finding is in strike contrast with other parasitic species such as M. tuberculosis where asparaginase is secreted out of the parasite...
March 15, 2018: International Journal of Biological Macromolecules
Mandy Berndt-Paetz, Luise Herbst, Annett Weimann, Andreas Gonsior, Jens-Uwe Stolzenburg, Jochen Neuhaus
PURPOSE: Muscarinic acetylcholine receptors (mAChRs) regulate a number of important physiological functions. Alteration of mAChR expression or function has been associated in the etiology of several pathologies including functional bladder disorders (e.g bladder pain syndrome/interstitial cystitis - BPS/IC). In a previous study we found specific mAChR expression patterns associated with BPS/IC, while correlation between protein and gene expression was lacking. Posttranslational regulatory mechanisms, e...
March 15, 2018: Acta Histochemica
Michael M Goodin
Protein interaction and localization studies in plants are a fundamental component of achieving mechanistic understanding of virus:plant interactions at the systems level. Many such studies are conducted using transient expression assays in leaves of Nicotiana benthamiana, the most widely used experimental plant host in virology, examined by laser-scanning confocal microscopy. This chapter provides a workflow for protein interaction and localization experiments, with particular attention to the many control and supporting assays that may also need to be performed...
2018: Advances in Virus Research
E Trybus, G Krol, T Obarzanowski, W Trybus, A Kopacz-Bednarska, M Obarzanowski, T Krol
Most studies on the effects of glucocorticosteroid therapy in rhinitis relate to their inhibitory effect on activation and the number of inflowing cells that are involved in the development and maintenance of inflammation. It is also very important to determine the range of effect of budesonide on residing cells (epithelial cells). The purpose of this study was to evaluate the effect of local budesonide therapy on the cytological image of the nasal mucosa, with attention paid to columnar cells in patients with rhinitis...
December 2017: Journal of Physiology and Pharmacology: An Official Journal of the Polish Physiological Society
Karl A Hujsak, Eric W Roth, William Kellogg, Yue Li, Vinayak P Dravid
Technological advances in electron microscopy, particularly improved detectors and aberration correctors, have led to higher throughput and less invasive imaging of materials and biological structures by enhancing signal-to-noise ratios at lower electron exposures. Analytical methods, such as electron energy loss spectroscopy (EELS) and energy dispersive x-ray spectrometry (EDS), have also benefitted and offer a rich set of local elemental and bonding information with nano-or atomic resolution. However, spatially resolved spectrum imaging with EELS and EDS continue to be difficult to scale due to limited detector collection angles or high signal background, requiring hours or even days for full maps...
March 10, 2018: Micron: the International Research and Review Journal for Microscopy
Kostantinos Kiakos, Bernhard Englinger, Stephanie K Yanow, Debora Wernitznig, Michael A Jakupec, Walter Berger, Bernhard K Keppler, John A Hartley, Moses Lee, Pravin C Patil
HxTfA 4 is a fluorescent analog of a potent cytotoxic and antimalarial agent, TfA 3, which is currently being investigated for the development of an antimalarial vaccine, PlasProtect®. HxTfA contains a p-anisylbenzimidazole or Hx moiety, which is endowed with a blue emission upon excitation at 318 nm; thus enabling it to be used as a surrogate for probing the cellular fate of TfA using confocal microscopy, and addressing the question of nuclear localization. HxTfA exhibits similar selectivity to TfA for A-tract sequences of DNA, alkylating adenine-N3, albeit at 10-fold higher concentrations...
March 5, 2018: Bioorganic & Medicinal Chemistry Letters
René Ledesma-Alonso, Romeli Barbosa, Jaime Ortegón
The characterization and reconstruction of heterogeneous materials, such as porous media and electrode materials, involve the application of image processing methods to data acquired by scanning electron microscopy or other microscopy techniques. Among them, binarization and decimation are critical in order to compute the correlation functions that characterize the microstructure of the above-mentioned materials. In this study, we present a theoretical analysis of the effects of the image-size reduction, due to the progressive and sequential decimation of the original image...
February 2018: Physical Review. E
Allison A Lindgren, Adam R Filipowicz, Julian B Hattler, Soon Ok Kim, Hye Kyung Chung, Marcelo J Kuroda, Edward M Johnson, Woong-Ki Kim
OBJECTIVE: HIV-1 infection of the brain and related cognitive impairment remain prevalent in HIV-1-infected subjects despite combination antiretroviral therapy. Sterile alpha motif and histidine-aspartate domain-containing protein 1 (SAMHD1) is a newly identified host restriction factor that blocks the replication of HIV-1 and other retroviruses in myeloid cells. Cell-cycle-regulated phosphorylation at residue Thr592 and viral protein X (Vpx)-mediated degradation of SAMHD1 have been shown to bypass SAMHD1 restriction in vitro...
March 15, 2018: AIDS
Frederik Flenner, Nicole Arlt, Mahtab Nasib, Sophie Schobesberger, Thea Koch, Ursula Ravens, Felix Friedrich, Viacheslav Nikolaev, Torsten Christ, Sebastian N Stehr
BACKGROUND: Systemic toxicity of local anesthetics is predominantly complicated by their myocardial toxicity. Especially long-acting local anesthetics exert a negative inotropic effect that has been described at lower concentrations than defined for blockade of myocardial ion channels. We evaluated the negative inotropic effect of bupivacaine at a concentration described for clinical toxicity testing the hypothesis that negative inotropy is a result of reduced Ca sensitivity rather than blockade of ion channels...
March 16, 2018: Anesthesiology
Maria Siemionow, Joanna Cwykiel, Ahlke Heydemann, Jesus Garcia, Enza Marchese, Krzysztof Siemionow, Erzsebet Szilagyi
Duchenne Muscular Dystrophy (DMD) is a progressive and lethal disease caused by mutations of the dystrophin gene. Currently no cure exists. Stem cell therapies targeting DMD are challenged by limited engraftment and rejection despite the use of immunosuppression. There is an urgent need to introduce new stem cell-based therapies that exhibit low allogenic profiles and improved cell engraftment. In this proof-of-concept study, we develop and test a new human stem cell-based approach to increase engraftment, limit rejection, and restore dystrophin expression in the mdx/scid mouse model of DMD...
March 15, 2018: Stem Cell Reviews
Beatriz Galatas, Helena Martí-Soler, Lidia Nhamussua, Pau Cisteró, Pedro Aide, Francisco Saute, Clara Menéndez, N Regina Rabinovich, Pedro L Alonso, Quique Bassat, Alfredo Mayor
Background: Afebrile Plasmodium falciparum infections usually remain undetected and untreated in the community and could potentially contribute to sustaining local malaria transmission in areas aiming for malaria elimination. Methods: Thirty-two men with afebrile P. falciparum infections detected with rapid diagnostic test (RDTs) were followed for 28 days. Kaplan-Meier estimates were computed to estimate probability of parasite positivity and of reducing parasitaemia by half of its initial level by day 28...
March 13, 2018: Clinical Infectious Diseases: An Official Publication of the Infectious Diseases Society of America
Amy M Bittel, Ashley M Davis, Lei Wang, Michel A Nederlof, Jorge O Escobedo, Robert M Strongin, Summer L Gibbs
Multicolor microscopy tools necessary to localize and visualize the complexity of subcellular systems are limited by current fluorophore technology. While commercial fluorophores cover spectral space from the ultraviolet to the near infrared region and are optimized for conventional bandpass based fluorescence microscopy, they are not ideal for highly multiplexed fluorescence microscopy as they tend to have short Stokes shifts, restricting the number of fluorophores that can be detected in a single sample to four to five...
March 15, 2018: Scientific Reports
Sangmoon Yoon, Hyobin Yoo, Seoung-Hun Kang, Miyoung Kim, Young-Kyun Kwon
We report the existence of latent order during core relaxation in the high-angle grain boundaries (GBs) of GaN films using atomic-resolution scanning transmission electron microscopy and ab initio density functional theory calculations. Core structures in the high-angle GBs are characterized by two pairs of Ga-N bonds located next to each other. The core type correlates strongly with the bond angle differences. We identify an order of core relaxation hidden in the high-angle GBs by further classifying the 5/7 atom cores into a stable 5/7 core (5/7(S)) and a metastable 5/7 core (5/7(M))...
March 15, 2018: Scientific Reports
Hisao Tsukamoto, Masahiro Higashi, Hideyoshi Motoki, Hiroki Watanabe, Christian Ganser, Koichi Nakajo, Yoshihiro Kubo, Takayuki Uchihashi, Yuji Furutani
Canonical K+ channels are tetrameric and highly K+ selective, whereas two-pore domain K+ (K2P) channels form dimers, but with a similar pore architecture. A two-pore domain potassium channel TWIK1 (KCNK1 or K2P1) allows permeation of Na+ and other monovalent ions, resulting mainly from the presence of Thr118 in the P1 domain. However, the mechanistic basis for this reduced selectivity is unclear. Using ion-exchange-induced difference infrared spectroscopy, we analyzed WT TWIK1 and T118I (highly K+ selective) and L228F (substitution in the P2 domain) TWIK1 variants and found that in the presence of K+ ions, WT and both variants exhibit an amide-I band at 1680 cm-1 This band corresponds to interactions of the backbone carbonyls in the selectivity filter with K+ , a feature very similar to that of the canonical K+ channel KcsA...
March 15, 2018: Journal of Biological Chemistry
P A Stapleton, C R McBride, J Yi, A B Abukabda, T R Nurkiewicz
Preconceptive health encompasses male and female reproductive capability. In females, this takes into account each of the stages of the estrous cycle. Microvascular reactivity varies throughout the estrous cycle in response to hormonal changes and in preparation for pregnancy. Microvascular alterations in response to engineered nanomaterial (ENM) exposure have been described within 24-hours of inhalation; however, the impact upon the uterine vasculature at differing estrous stages and at late-stage pregnancy is unclear...
March 12, 2018: Reproductive Toxicology
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