scholarly journals Nuclear pores as versatile reference standards for quantitative superresolution microscopy

2019 ◽  
Author(s):  
Jervis Vermal Thevathasan ◽  
Maurice Kahnwald ◽  
Konstanty Cieśliński ◽  
Philipp Hoess ◽  
Sudheer Kumar Peneti ◽  
...  
Keyword(s):  
Cell Lines ◽  
Fluorescent Protein ◽  
Reference Standards ◽  
Nuclear Pore ◽  
Biologically Relevant ◽  
Superresolution Microscopy ◽  
Absolute Measurements ◽  
Quantitative Fluorescence

AbstractQuantitative fluorescence and superresolution microscopy are often limited by insufficient data quality or artifacts. In this context, it is essential to have biologically relevant control samples to benchmark and optimize the quality of microscopes, labels and imaging conditions.Here we exploit the stereotypic arrangement of proteins in the nuclear pore complex as in situ reference structures to characterize the performance of a variety of microscopy modalities. We created four genome edited cell lines in which we endogenously labeled the nucleoporin Nup96 with mEGFP, SNAP-tag or HaloTag or the photoconvertible fluorescent protein mMaple. We demonstrate their use a) as 3D resolution standards for calibration and quality control, b) to quantify absolute labeling efficiencies and c) as precise reference standards for molecular counting.These cell lines will enable the broad community to assess the quality of their microscopes and labels, and to perform quantitative, absolute measurements.

scholarly journals The BBSome assembly is spatially controlled by BBS1 and BBS4 in human cells

2020 ◽  
Vol 295 (42) ◽  
pp. 14279-14290 ◽  
Author(s):  
Avishek Prasai ◽  
Marketa Schmidt Cernohorska ◽  
Klara Ruppova ◽  
Veronika Niederlova ◽  
Monika Andelova ◽  
...  
Keyword(s):  
Cell Lines ◽  
Primary Cilia ◽  
Fluorescent Protein ◽  
Correlation Spectroscopy ◽  
Bardet Biedl Syndrome ◽  
Fluorescence Correlation ◽  
Biochemical Assays ◽  
Genes Encoding ◽  
Microscopy Techniques ◽  
Quantitative Fluorescence

Bardet–Biedl syndrome (BBS) is a pleiotropic ciliopathy caused by dysfunction of primary cilia. More than half of BBS patients carry mutations in one of eight genes encoding for subunits of a protein complex, the BBSome, which mediates trafficking of ciliary cargoes. In this study, we elucidated the mechanisms of the BBSome assembly in living cells and how this process is spatially regulated. We generated a large library of human cell lines deficient in a particular BBSome subunit and expressing another subunit tagged with a fluorescent protein. We analyzed these cell lines utilizing biochemical assays, conventional and expansion microscopy, and quantitative fluorescence microscopy techniques: fluorescence recovery after photobleaching and fluorescence correlation spectroscopy. Our data revealed that the BBSome formation is a sequential process. We show that the pre-BBSome is nucleated by BBS4 and assembled at pericentriolar satellites, followed by the translocation of the BBSome into the ciliary base mediated by BBS1. Our results provide a framework for elucidating how BBS-causative mutations interfere with the biogenesis of the BBSome.

Pengaruh Jenis Co-Solvent terhadap Rendemen dan Mutu Biodiesel secara Transesterifikasi In Situ Menggunakan Radiasi Gelombang Mikro

2016 ◽  
Vol 10 (2) ◽  
pp. 119-126
Author(s):  
Mahlinda Mahlinda ◽  
Fitriana Djafar
Keyword(s):  
Catalyst Concentration ◽  
Maximum Yield ◽  
Acid Value ◽  
Experimental Result ◽  
In Situ Transesterification ◽  
Yield And Quality ◽  
Solvent Type ◽  
Biodiesel Quality

The main purpose of this research was to observer effect co-solvent type (n-Hexane, chloroform and without co-solvent)  toward yield and quality of biodiesel via in situ transesterification process using microwave irradiation. The process was studied at microwave power 450 watt, reaction time 4 minutes, methanol to seed ratio 25:1 and catalyst concentration 5%. The physicochemical parameters of the biodiesel produced such as viscosity, density and acid value were analysed and compared with the SNI 7182-2012 standard. The experimental result showed the maximum yield biodiesel 78,32% obtained by using co-solvent chloroform.Test result of physicochemical properties (viscosity, density and acid value) of biodiesel products using co solvent n-Hexane, chloroform and without co solvent showed that these products conform to the SNI 7182-2012 standars. The type of co-solvent only affectedon biodiesel yield dan not affected on biodiesel quality (viscosity, density and acid value).  ABSTRAKTujuan penelitian ini adalah untuk mempelajari pengaruh jenis co-solvent (n-Hexane, chloroform dan tanpa co-solvent) terhadap rendemen dan mutu biodiesel secara trasesterifikasi in situ menggunakan radiasi gelombang mikro. Proses dilakukan pada daya gelombang mikro 450 watt, waktu reaksi 4 menit, perbandingan berat metanol terhadap bahan baku 25:1 dan jumlah katalis 5%. Parameter fisiko kimia dari produk biodiesel seperti viskositas, densitas dan angka asam di analisa dan dibandingkan dengan standar SNI 7182-2012 tentang biodiesel. Hasil penelitian menunjukkan rendemen maksimum biodiesel sebesar 78,32% diperoleh dengan menggunakan co-solvent chloroform. Hasil pengujian  karakteristik fisiko kimia (viskositas, densitas dan angka asam) dari produk biodiesel menggunakan co-solvent n-Hexane, chloroform dan tanpa co-solvent menunjukkan bahwa semua parameter ini masih memenuhi standar SNI 1782-2012 tentang biodiesel. Jenis co-solvent hanya berpengaruh pada rendemen biodiesel dan tidak berpengaruh terhadap mutu biodiesel (viskositas, densitas dan bilangan asam).Kata kunci: co-solvent, in situ transesterifikasi, microwave, rendemen, mutu   

Estudio del proceso de lavado de contenedores para eliminar tiempos y movimientos innecesarios, caso: Wash Containers S.A de C.V.

2020 ◽  
pp. 25-36
Author(s):  
Juan Alfredo Lino-Gamiño ◽  
Carlos Méndez-González ◽  
Eduardo José Salazar-Araujo ◽  
Pablo Adrián Magaña-Sánchez
Keyword(s):  
Business Process ◽  
Value Chain ◽  
Process Management ◽  
Core Business ◽  
The Core ◽  
Washing Process ◽  
Overall Performance ◽  
The Times

In the value chain it is important to keep in mind the core business of the company, since it depends largely on the competitiveness of the company and its overall performance, bearing in mind that all business indicators depend on it. In this work we will study the washing process within the company WASH CONTAINERS SA DE CV, to improve the washing processes and in this way reduce times and movements in the process leading the company to reduce costs considerably within the operations company daily, having a more competitive operation and with greater profit margin in its business process. Goals: It Improve the logistics of the movement of containers for washing and with it the core business of the company. Methodology: The action research will be applied applying Business Process Management for the improvement of processes in situ, it will be developed in a certain period of time and with that it will establish an improvement projection. Contribution: The improvement of the times for the disposal of the containers and their subsequent use, allows a better competitiveness and with it the income of the company, on the other hand, the transport companies improve in performance in quantity, quality of disposition and with it their income.

Interphase Quantitative Fluorescence in Situ Hybridization (IQ-FISH)

2019 ◽  
Vol 3 (1) ◽  
Author(s):  
Ivan Y. Iourov ◽  
◽  
Ilia V. Soloviev ◽  
Yuri B. Yurov ◽  
Svetlana G. Vorsanova ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Fluorescence In Situ Hybridization ◽  
Quantitative Fluorescence

scholarly journals Mex67p of Schizosaccharomyces pombeInteracts with Rae1p in Mediating mRNA Export

2000 ◽  
Vol 20 (23) ◽  
pp. 8767-8782 ◽  
Author(s):  
Jin Ho Yoon ◽  
Dona C. Love ◽  
Anjan Guhathakurta ◽  
John A. Hanover ◽  
Ravi Dhar
Keyword(s):  
Nuclear Export ◽  
Rna Binding ◽  
Fluorescent Protein ◽  
Synthetic Lethality ◽  
Sequence Similarity ◽  
Mrna Export ◽  
Nuclear Periphery ◽  
Nuclear Pore ◽  
Multicopy Suppressor ◽  
Accessory Factor

ABSTRACT We identified the Schizosaccharomyces pombe mex67 gene (spmex67) as a multicopy suppressor of rae1-167 nup184-1 synthetic lethality and the rae1-167 tsmutation. spMex67p, a 596-amino-acid-long protein, has considerable sequence similarity to the Saccharomyces cerevisiae Mex67p (scMex67p) and human Tap. In contrast toscMEX67, spmex67 is essential for neither growth nor nuclear export of mRNA. However, an spmex67 null mutation (Δmex67) is synthetically lethal with therae1-167 mutation and accumulates poly(A)+ RNA in the nucleus. We identified a central region (149 to 505 amino acids) within spMex67p that associates with a complex containing Rae1p that complements growth and mRNA export defects of therae1-167 Δmex67 synthetic lethality. This region is devoid of RNA-binding, N-terminal nuclear localization, and the C-terminal nuclear pore complex-targeting regions. The (149–505)-green fluorescent protein (GFP) fusion is found diffused throughout the cell. Overexpression of spMex67p inhibits growth and mRNA export and results in the redistribution of the diffused localization of the (149–505)-GFP fusion to the nucleus and the nuclear periphery. These results suggest that spMex67p competes for essential mRNA export factor(s). Finally, we propose that the 149–505 region of spMex67p could act as an accessory factor in Rae1p-dependent transport and that spMex67p participates at various common steps with Rae1p export complexes in promoting the export of mRNA.

scholarly journals Modelling the Quality of Bathing Waters in the Adriatic Sea

Water ◽  
2021 ◽  
Vol 13 (11) ◽  
pp. 1525
Author(s):  
Christian Ferrarin ◽  
Pierluigi Penna ◽  
Antonella Penna ◽  
Vedrana Spada ◽  
Fabio Ricci ◽  
...  
Keyword(s):  
Adriatic Sea ◽  
Management Tool ◽  
Negative Consequences ◽  
Sea Temperature ◽  
Bathing Water Directive ◽  
Bathing Waters ◽  
Diffusion Dynamics ◽  
Study Sites

The aim of this study is to develop a relocatable modelling system able to describe the microbial contamination that affects the quality of coastal bathing waters. Pollution events are mainly triggered by urban sewer outflows during massive rainy events, with relevant negative consequences on the marine environment and tourism and related activities of coastal towns. A finite element hydrodynamic model was applied to five study areas in the Adriatic Sea, which differ for urban, oceanographic and morphological conditions. With the help of transport-diffusion and microbial decay modules, the distribution of Escherichia coli was investigated during significant events. The numerical investigation was supported by detailed in situ observational datasets. The model results were evaluated against water level, sea temperature, salinity and E. coli concentrations acquired in situ, demonstrating the capacity of the modelling suite in simulating the circulation in the coastal areas of the Adriatic Sea, as well as several main transport and diffusion dynamics, such as riverine and polluted waters dispersion. Moreover, the results of the simulations were used to perform a comparative analysis among the different study sites, demonstrating that dilution and mixing, mostly induced by the tidal action, had a stronger effect on bacteria reduction with respect to microbial decay. Stratification and estuarine dynamics also play an important role in governing microbial concentration. The modelling suite can be used as a beach management tool for improving protection of public health, as required by the EU Bathing Water Directive.

scholarly journals Chlorosulfuric acid-assisted production of functional 2D materials

2021 ◽  
Vol 5 (1) ◽  
Author(s):  
Mohsen Moazzami Gudarzi ◽  
Maryana Asaad ◽  
Boyang Mao ◽  
Gergo Pinter ◽  
Jianqiang Guo ◽  
...  
Keyword(s):  
Hexagonal Boron Nitride ◽  
2D Materials ◽  
Real Life ◽  
Large Area ◽  
Polar Solvents ◽  
Aspect Ratios ◽  
Two Dimensional Materials ◽  
Liquid Phase Exfoliation

AbstractThe use of two-dimensional materials in bulk functional applications requires the ability to fabricate defect-free 2D sheets with large aspect ratios. Despite huge research efforts, current bulk exfoliation methods require a compromise between the quality of the final flakes and their lateral size, restricting the effectiveness of the product. In this work, we describe an intercalation-assisted exfoliation route, which allows the production of high-quality graphene, hexagonal boron nitride, and molybdenum disulfide 2D sheets with average aspect ratios 30 times larger than that obtained via conventional liquid-phase exfoliation. The combination of chlorosulfuric acid intercalation with in situ pyrene sulfonate functionalisation produces a suspension of thin large-area flakes, which are stable in various polar solvents. The described method is simple and requires no special laboratory conditions. We demonstrate that these suspensions can be used for fabrication of laminates and coatings with electrical properties suitable for a number of real-life applications.

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