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Universal NicE-Seq: A Simple and Quick Method for Accessible Chromatin Detection in Fixed Cells.

Genome-wide accessible chromatin sequencing and identification has enabled deciphering the epigenetic information encoded in chromatin, revealing accessible promoters, enhancers, nucleosome positioning, transcription factor occupancy, and other chromosomal protein binding. The starting biological materials are often fixed using formaldehyde crosslinking. Here, we describe accessible chromatin library preparation from low numbers of formaldehyde-crosslinked cells using a modified nick translation method, where a nicking enzyme nicks one strand of DNA and DNA polymerase incorporates biotin-conju…

Lateral Flow Recombinase Polymerase Amplification Assays for the Detection of Human Plasmodium Species.

This study highlights the development of two lateral flow recombinase polymerase amplification assays for the diagnosis of human malaria. The lateral flow cassettes contained test lines that captured biotin-, 6-carboxyfluorescein, digoxigenin-, cyanine 5-, and dinitrophenyl-labeled amplicons. The overall process can be completed in 30 minutes. Recombinase polymerase amplification coupled with lateral flow had a detection limit of 1 copy/µL for Plasmodium knowlesi, Plasmodium vivax, and Plasmodium falciparum. No cross-reactivity was observed among nonhuman malaria parasites such as Plasmodium c…

Apoptosis of periodontal ligament cells induced by mechanical stress during tooth movement.

The mechanical force generated during tooth movement creates compressed and cell-free areas in the periodontal membrane. The way in which periodontal ligament cells disappear at the compressed area during tooth movement remains unclear. In the present study we examined whether periodontal ligament cells undergo apoptosis by mechanical stress during tooth movement using the terminal deoxynucleotidyl transferase (TdT)-mediated dUTP-biotin nick-end-labelling method (TUNEL). TUNEL-positive stainings of periodontal ligament cells began to appear at the compressed areas 12 h after tooth movement, an…

A case of secondary syphilis with mucous patches on the hard palate.

A rare case of secondary syphilis showing mucous patches on the hard palate is reported. A 31-year-old male had two erosive patches which were slightly raised on his hard palate. A linear lesion was also present on the inside of his right alveolar process. Many red-brown macules on his abdomen and bilateral inguinal lymphadenopathy accompanied these symptoms. The serological tests for syphilis were positive: VDRL test 1:512, TPHA test 1:20,480, and IgM-FTA-ABS test 1:40. Immunohistochemical staining with rabbit monoclonal antibody to Treponema pallidum by the biotin-streptoavidin system detect…

Detection of human papillomavirus DNA sequences in cervical lesions by in situ hybridization using biotinylated DNA probes.

Ninety-six biopsy specimens were collected for studying the association of specific sequences of HPV DNA with genital warts, precancerous lesions and invasive cervical cancer. Using DNA probes labeled with biotin, 17 of the 22 (77.27%) specimens showing condyloma and 1 of the 14 (7.14%) specimens displaying subclinical papillomavirus infection (SPI) of cervix were found to be positive for HPV 6/11; 4 of 23 (17.39%) specimens of squamous epithelial cervical cancer and 2 of the 23 (8.70%) specimens of cervical intraepithelial neoplasia (CIN) were positive for HPV 16/18. All the 14 specimens of c…

Методика определения поверхности чувствительности для оптимизации выражения CHIKV-SFv-BAD против E2 в Escherichia coli Origami B (DE3) для обнаружения вируса Чикунгунья.

Вирус Чикунгунья (CHIKV) является патогеном, передаваемым через комары, который несет ответственность за повторяющиеся вспышки, что подчеркивает потребность в оперативных и точных диагностических инструментах. Гликпротеин E2 CHIKV является перспективной целью для обнаружения на базе антител. Цель этого исследования заключается в определении оптимальных условий выражения рекомбинированного одноцепного изменяющегося фрагмента (scFv), сплавленного с доменом принятия биотинов (BAD), специфичным для CHIKV E2 glycoprotein в Escherichia coli Origami B (DE3). Оптимизация была произведена с использован

Совпадение в аннотации оригинала
Chikungunya virus (CHIKV) is a mosquito-borne pathogen responsible for recurring outbreaks, highlighting the need for rapid and accurate dia … e the optimal expression conditions of recombinant single-chain variable fragment (scFv) fused with biotin acceptor domain (BAD) specific to the CHIKV E2 glycoprotein in Escherichia coli Origami B (DE3). The optimization was performed using the Box-Behnken design of response surface methodology (RSM), with the IPTG inducer concentration, induction time, …

Insights into the neuroprotective mechanisms of 2-iminobiotin employing an in-vitro model of hypoxic-ischemic cell injury.

Several animal models have been used to simulate cerebral hypoxia-ischemia and suggested neuroprotective effects of the biotin analogue 2-iminobiotin (2-IB). The aims of this study were to employ a human in-vitro hypoxia model to confirm protective effects of 2-IB on neuronal cells, determine the optimal neuroprotective concentrations of 2-IB and scrutinize underlying cellular effects of 2-IB. Neuronal IMR-32 cells were exposed to hypoxia employing an enzymatic hypoxia system and were thereafter incubated with various concentrations of 2-IB (10 to 300ng/ml). Cell damage, metabolic activity and…

Increasing the efficacy of bioorthogonal click reactions for bioconjugation: a comparative study.

Raising the bar: the efficacy of bioorthogonal reactions for bioconjugation has been thoroughly evaluated in four different biological settings. Powered by the development of new biocompatible ligands, the copper-catalyzed azide-alkyne cycloaddition has brought about unsurpassed bioconjugation efficiency, and thus it holds great promise as a highly potent and adaptive tool for a broader spectrum of biological applications.

A new, simple method for linking of antibodies to atomic force microscopy tips.

Functionalization of atomic force microscope (AFM) tips with bioligands converts them into monomolecular biosensors which can detect complem … sine residues of proteins via Schiff base formation. The method has been critically examined, using biotinylated IgG as bioligand on the tip and mica-bound avidin as complementary receptor. These experiments were well reproduced on amino-functionalized silicon nitride chips where the number of specifically bound IgG molecules (approximately 2000 per micr …

Mitochondrial function and toxicity: role of the B vitamin family on mitochondrial energy metabolism.

The B vitamins are water-soluble vitamins required as coenzymes for enzymes essential for cell function. This review focuses on their essent … ntial for alpha-ketoglutarate and pyruvate dehydrogenase complexes as well as fatty acid oxidation. Biotin (B7) is the coenzyme of decarboxylases required for gluconeogenesis and fatty acid oxidation. Pyridoxal (B6), folate and cobalamin (B12) properties are reviewed elsewhere in this issue. The experimental animal and clinical evidence that vitamin B th …

Fiber connections of the central nucleus of semicircular torus in cyprinids.

Fiber connections of the presumed auditory portion of the semicircular torus or the central nucleus (TSc) as well as other central and peripheral auditory structures were studied by tract-tracing methods in carp and goldfish. Major ascending projections to the TSc originated from the descending octaval nucleus (DO) and medullary secondary octaval population (SO). Toropetal neurons of the DO were within the saccular terminal zone. A small number of toropetal DO neurons might receive inputs from other inner ear and lateral line endorgans as well. Fibers from the DO terminated in a deep zone of t

Acute in vivo proximity labeling for membrane-targeted proteomics in neuronal circuits.

A major goal within molecular systems neuroscience is to bridge the study of neuronal circuit function with changes in protein expression and localization in awake behaving animals. However, there are limited tools for capturing changes in subcellularly defined proteomes within neuronal circuits during activity-gated timescales in vivo. Here, we engineered targeted versions of the proximity labeling enzyme TurboID to tag proteins at the neuronal membrane during a user-delivered biotin injection. We optimized a labeling strategy that enables a 1- to 2-h labeling window and tagged proteins in me…

CCT8 associates with the MYO10 motor domain and regulates filopodia and breast cancer cell invasion.

Cancer cells utilize filopodia to explore, adhere to and invade their surrounding microenvironment, yet the protein networks that organize these protrusions remain incompletely defined. To uncover the molecular machinery underlying MYO10-positive filopodia, we targeted the fast biotin ligase TurboID to the motor protein MYO10. Proximity biotinylation in two cell types revealed hundreds of potential MYO10 interactors. A targeted microscopy and siRNA screen identified MINK1, SCRIB, CSNK1A1 and CCT8 as new regulators of MYO10-mediated filopodia formation. Focusing on one of the interactors shared…

Sensitive, Low-Background-Signal miRNA Analysis via the Self-Priming-Initiated Color Reaction Loaded on a Rolling Circle Amplification Product.

MicroRNAs (miRNAs) have been regarded as potential biomarkers in evaluating various diseases, such as pregnancy-induced hypertension and cancers. However, sensitive and reliable miRNA detection is still a challenge due to the low amplification efficiency and high background signal. Herein, we developed a colorimetric method for miRNA detection utilizing the self-priming-initiated color reaction loaded on a rolling circle amplification (RCA) product. In this method, a biotin-labeled RCA product is fixed on the surface of the streptavidin-coated wells, and the interfering components in samples a…

Chemical proteomics tracks virus entry and uncovers NCAM1 as Zika virus receptor.

The outbreak of Zika virus (ZIKV) in 2016 created worldwide health emergency which demand urgent research efforts on understanding the virus biology and developing therapeutic strategies. Here, we present a time-resolved chemical proteomic strategy to track the early-stage entry of ZIKV into host cells. ZIKV was labeled on its surface with a chemical probe, which carries a photocrosslinker to covalently link virus-interacting proteins in living cells on UV exposure at different time points, and a biotin tag for subsequent enrichment and mass spectrometric identification of the receptor or othe…

A colorimetric aptamer-based method for detection of cadmium using the enhanced peroxidase-like activity of Au-MoS2 nanocomposites.

In this work, a colorimetric aptamer-based method for detection of cadmium using gold nanoparticles modified MoS2 nanocomposites as enzyme mimic is established. In short, biotinylated Cd2+ aptamers are immobilized by biotin-avidin binding on the bottoms of the microplate, the complementary strands of Cd2+ aptamers are connected to the Au-MoS2 nanocomposites which have the function of enhanced peroxidase-like activity. The csDNA-Au-MoS2 signal probe and target Cd2+ compete for binding Cd2+ aptamer, the color change can be observed by addition of chromogenic substrate, thereby realizing visual d…

A quantitative enzyme-linked immunoassay (ELISA) to approximate complement-fixing antibody titers in serum from patients with coccidioidomycosis.

Coccidioidomycosis is most frequently diagnosed serologically, and the quantitative test for complement-fixing antibodies is considered prog … onformational. Further, anchoring the antigenic peptide to the ELISA plate by means of a C-terminal biotin-mimic peptide tag instead of allowing the peptide to randomly adhere to the plastic plate improves sensitivity of antibody detection by 1-2 logs in different sera. The newly developed ELISA shows a significant quantitative correlation with complemen …

Ligand-Appended BBB-Targeted Nanocarriers (LABTNs).

Delivering therapeutics across the blood brain barrier (BBB) remains the rate-limiting step in brain medicine research. Three main categorie … l as recent advances in BBB targeting tactics including antibody-directed enzyme prodrug therapy, a biotin-avidin conjugated system, and chimeric peptides. This review serves as an excellent source of knowledge for budding brain researchers.

[Expression of ERK and nm23-H1 in tongue squamous cell carcinoma and its relation with invasion and metastasis].

PURPOSE: To reveal the relations between expression of extracellular signal-regulated kinase (ERK) and nm23-H1 and tumor invasion and metastasis in tongue squamous cell carcinoma (TSCC). METHODS: The expression of ERK and nm23-H1 at protein level was examined in 74 cases of TSCC by strep avidin-biotin complex (SABC) immunohistochemical technique. SPSS10.0 software package was used for data analysis. RESULTS: The expression of ERK was positively related to TNM clinical stage and cervical lymph node metastasis. The expression of nm23-H1 was negatively related to TNM clinical stage and cervical…

Identification of host cell factors required for intoxication through use of modified cholera toxin.

We describe a novel labeling strategy to site-specifically attach fluorophores, biotin, and proteins to the C terminus of the A1 subunit (CTA1) of cholera toxin (CTx) in an otherwise correctly assembled and active CTx complex. Using a biotinylated N-linked glycosylation reporter peptide attached to CTA1, we provide direct evidence that ~12% of the internalized CTA1 pool reaches the ER. We also explored the sortase labeling method to attach the catalytic subunit of diphtheria toxin as a toxic warhead to CTA1, thus converting CTx into a cytolethal toxin. This new toxin conjugate enabled us to co…