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During mitosis, kinetochore-microtubule attachments are monitored by a molecular surveillance system known as the spindle assembly checkpoint. The prevailing model posits that dynein evicts checkpoint proteins (e.g., Mad1, Mad2) from stably attached kinetochores by transporting them away from kinetochores, thus contributing to checkpoint silencing. However, the mechanism by which dynein performs this function, and its precise role in checkpoint silencing remain unresolved. Here, we find that dynein's role in checkpoint silencing is restricted to evicting checkpoint effectors from the fibrous corona, and not the outer kinetochore. Dynein evicts these molecules from the corona in a manner that does not require stable, end-on microtubule attachments. Thus, by disassembling the corona through indiscriminate microtubule encounters, dynein primes the checkpoint signaling apparatus so it can respond to stable end-on microtubule attachments and permit cells to progress through mitosis. Accordingly, we find that dynein function in checkpoint silencing becomes largely dispensable in cells in which checkpoint effectors are excluded from the corona.

Antibodies are indispensable tools used for a large number of applications in both foundational and translational bioscience research; however, there are drawbacks to using traditional antibodies generated in animals. These include a lack of standardization leading to problems with reproducibility, high costs of antibodies purchased from commercial sources, and ethical concerns regarding the large number of animals used to generate antibodies. To address these issues, we have developed practical methodologies and tools for generating low-cost, high-yield preparations of recombinant monoclonal antibodies and antibody fragments directed to protein epitopes from primary sequences. We describe these methods here, as well as approaches to diversify monoclonal antibodies, including customization of antibody species specificity, generation of genetically encoded small antibody fragments, and conversion of single chain antibody fragments (e.g. scFv) into full-length, bivalent antibodies. This study focuses on antibodies directed to epitopes important for mitosis and kinetochore function; however, the methods and reagents described here are applicable to antibodies and antibody fragments for use in any field., Competing Interests: KD, JM, AI, WL, LS, KS, SA, NZ, TS, DV, JN No competing interests declared, JD Reviewing editor, eLife, (© 2021, DeLuca et al.)

Waste of Mytella falcata shell was used as low-cost adsorbent to remove the biocide Basic Green 4 (BG4) from water. Shells were collected form trash nearby the lagoon were Mytella falcata is fished. After clean, dry and crushed, the powder was characterized by scanning electron microscopy (SEM), Fourier-transform infrared spectroscopy (FT-IR), X-ray powder diffraction (XRD), and X-ray dispersive energy spectroscopy (EDS). Both kinetic and equilibrium adsorption tests are carried out. Adsorbent regenerability was tested during adsorption/desorption cycles, using a UV photo-regeneration process. The maximum adsorption capacity reached 539.24 mg.g-1 (60 °C), which was higher than those retrieved for other materials with similar origin. The kinetic results indicated that the process followed pseudo-second order model. Equilibrium data indicate an increase in BG4 adsorption capacity with temperature and Sips model had better fit for all the investigated temperatures (30, 40, 50 and 60 °C). The regeneration/reuse test indicated that the adsorbent is able to assure a BG4 removal above 70 % during five adsorption/desorption cycles evaluated. Thermodynamic parameters suggested that adsorption is spontaneous, endothermal, governed by chemisorption and with structural changes in the solid surface upon adsorption., (Copyright © 2020 Elsevier B.V. All rights reserved.)

A new hybrid microextraction technique (hollow fiber microextraction) is presented that uses the main concepts and advantages of the modern miniaturized devices used for trace analysis. This novel analytical approach uses devices made of polypropylene membranes (10.0 mm long and 0.6 mm internal diameter) in which convenient organic solvents are embedded that promote fast kinetics during the enrichment process, using the floating sampling technology concept. An innovative analytical cycle is also proposed by use of low-cost disposable devices during the microextraction stage together with a user-friendly ("single liquid desorption step") back-extraction stage in compliance with green analytical chemistry principles. To evaluate the performance of the proposed technique, 18 polycyclic aromatic hydrocarbons (PAHs) were used as model compounds and were monitored by gas chromatography coupled with mass spectrometry. Under optimized experimental conditions, assays performed on 25 mL aqueous samples spiked with the PAHs at trace level yielded average recoveries between (14.5 ± 8.2)% (dibenzo[a,h]anthracene) and (90.4 ± 8.4)% (benzo[a]anthracene) with use of a device in which n-nonane had been embedded. Low detection limits were also achieved (2.50-6.00 ng L -1 ), as well as good linear dynamic ranges (20.00-2000.00 ng L -1 ), with suitable coefficients of determination (r 2 > 0.9905) and appropriate precision (relative standard deviation below 15%). By use of the standard addition method, the proposed hybrid microextraction technique had remarkable performance to monitor PAHs at the ultratrace level in several types of matrices, including surface water, wastewater, soil, tea, and fish liver samples. From the data obtained, the new hybrid hollow fiber microextraction technique proved to be user-friendly, eco-friendly, cost-effective, and very competitive for routine work. In short, the novel microextraction technique proposed herein is a remarkable alternative to other well-established microextraction techniques for ultratrace analysis of emerging compounds in real matrices. Graphical abstract Innovative analytical procedure for hollow fiber microextraction (HFμE). GC gas chromatography, LD liquid desorption.

We aimed to evaluate the ecological integrity of a large river, which receives agricultural and urban effluents and is used to water public supply. The fish species Astyanax bifasciatus was used as bioindicator during winter and spring 2012, and summer 2013 at the Middle Iguaçu River basin in Paraná state, Brazil. Water chemical and physical measures and ecotoxicological tests were carried out as well biochemical and genetic biomarkers in sampled fish in each period. The studied area was divided in three sample points: SP1, located where the water is collected to public supply; SP2, located in an urbanized area, and SP3, located at an urbanized area with the discharge of the sewage treatment. Although water chemical and physical analyzes were range of the Brazilian law to hydric bodies, anticholinesterasic effects were found in winter, oxidative stress in summer and spring. The higher genotoxic effect was in winter to all sample points. The temporal variation in biomarkers and the detection of caffeine in the water call attention to the water quality in this river mainly to be used to public supply., (Copyright © 2015 Elsevier Ltd. All rights reserved.)