RESUMEN
Antibodies are one of the most important reagents used in biomedical and fundamental research, used to identify, and quantify proteins, contribute to knowledge of disease mechanisms, and validate drug targets. Yet many antibodies used in research do not recognize their intended target, or recognize additional molecules, compromising the integrity of research findings and leading to waste of resources, lack of reproducibility, failure of research projects, and delays in drug development. Researchers frequently use antibodies without confirming that they perform as intended in their application of interest. Here we argue that the determinants of end-user antibody choice and use are critical, and under-addressed, behavioral drivers of this problem. This interacts with the batch-to-batch variability of these biological reagents, and the paucity of available characterization data for most antibodies, making it more difficult for researchers to choose high quality reagents and perform necessary validation experiments. The open-science company YCharOS works with major antibody manufacturers and knockout cell line producers to characterize antibodies, identifying high-performing renewable antibodies for many targets in neuroscience. This shows the progress that can be made by stakeholders working together. However, their work so far applies to only a tiny fraction of available antibodies. Where characterization data exists, end-users need help to find and use it appropriately. While progress has been made in the context of technical solutions and antibody characterization, we argue that initiatives to make best practice behaviors by researchers more feasible, easy, and rewarding are needed. Global cooperation and coordination between multiple partners and stakeholders will be crucial to address the technical, policy, behavioral, and open data sharing challenges. We offer potential solutions by describing our Only Good Antibodies initiative, a community of researchers and partner organizations working toward the necessary change. We conclude with an open invitation for stakeholders, including researchers, to join our cause.
Asunto(s)
Anticuerpos , Difusión de la Información , Reproducibilidad de los Resultados , Línea Celular , PolíticasRESUMEN
The transient receptor potential cation channel family member ankyrin 1 (TRPA1) is a potential target for several diseases, but detection of human TRPA1 (hTRPA1) protein in cells and tissues is problematic as rigorous antibody validation is lacking. We expressed hTRPA1 in a TRPA1-negative cell line to evaluate 5 commercially available antibodies by western blotting, immunofluorescence, immunocytochemistry and flow cytometry. The three most cited anti-TRPA1 antibodies lacked sensitivity and/or specificity, but two mouse monoclonal anti-TRPA1 antibodies detected hTRPA1 specifically in the above assays. This enabled the development of a flow cytometry assay, which demonstrated strong expression of TRPA1 in human lung myofibroblasts, human airway smooth muscle cells but not lung mast cells. The most cited anti-TRPA1 antibodies lack sensitivity and/or specificity for hTRPA1. We have identified two anti-TRPA1 antibodies which detect hTRPA1 specifically. Previously published data regarding human TRPA1 protein expression may need revisiting.
Asunto(s)
Anticuerpos/química , Miocitos del Músculo Liso/metabolismo , Miofibroblastos/metabolismo , Canal Catiónico TRPA1/metabolismo , Calcio/metabolismo , Señalización del Calcio , Epítopos/química , Citometría de Flujo , Perfilación de la Expresión Génica , Regulación de la Expresión Génica , Proteínas Fluorescentes Verdes/metabolismo , Células HEK293 , Humanos , Inmunohistoquímica , Pulmón/citología , Microscopía FluorescenteRESUMEN
Radon, helium and uranium measurements have been carried out in hot water springs in the Parbati and Beas valleys of Himachal Himalaya. Most of these hot springs are known as famous pilgrimage centers. The activity of dissolved radon in the liquid phase is found to vary widely, by an order of magnitude, between 10 and 750 Bq L(-1), whereas, the dissolved helium content in these thermal springs varies between 10 and 100 ppm. The uranium contents are low and vary from <0.01 to 5 microg L(-1). The measured values of radon, helium and uranium are possibly controlled by structural geology, namely the presence of pervious fault systems, and by the lithology of the leached host rocks. Redox-potential geochemical barriers cause the mobilization of uranile ions in solution (UO2+); the most plausible hypothesis is when the conditions are oxidising, confirming the importance of physico-chemical conditions up to the supergenic environment, to control the fluid geochemistry of the U-He-222Rn system. Some evidence is available from both geothermometric considerations and geochemical data which will be reported elsewhere, whereas the present study is focused on U decay series-noble gas geochemistry. The first analysis of collected 3He/4He data is consistent with a crustal signature at the studied thermal springs.
Asunto(s)
Helio/análisis , Manantiales de Aguas Termales , Contaminantes Radiactivos/análisis , Radón/análisis , Uranio/análisis , Recolección de Datos , Monitoreo del Ambiente , Fenómenos Geológicos , Geología , Helio/química , India , Gases Nobles/análisis , Gases Nobles/química , Oxidación-Reducción , Radón/química , Uranio/químicaRESUMEN
Radon measurements have been carried out in groundwater of Himachal Pradesh and Punjab states, India. Radon concentration values in potable water show a wide range of variation from source to source and from place to place. Generally, radon concentration values in thermal springs groundwater have been found to be higher than the values from other sources.
Asunto(s)
Radón/análisis , Abastecimiento de Agua , Monitoreo del Ambiente , India , SueloRESUMEN
It is well established that some areas of Himachal Pradesh (H.P.) state of India situated in the environs of the Himalayan mountains are relatively rich in uranium-bearing minerals. Some earlier studies by our group have indicated high levels of radon (>200 Bq m(-3)) in the dwellings. It is in this context that an indoor radon/thoron survey has been carried out in selected villages of four districts in the state of H.P. This survey has been conducted as a part of a national, coordinated project using twin chamber dosemeter cups designed by the Environmental Assessment Division (EAD), Department of Atomic Energy, Govt. of India. The track-etch technique is used for calibration of plastic detector LR-115 type-II which are employed for recording alpha tracks due to radon/thoron and their daughters. Year long radon/thoron data have been collected for seasonal correlations of indoor radon/thoron in the dwellings. The indoor radon levels have been found to vary from a minimum value of 17.4 Bq m(-3) to a maximum value of 140.3 Bq m(-3). The indoor thoron levels vary from a minimum value of 5.2 Bq m(-3) to a maximum value of 131.9 Bq m(-3). The year average dose rate for the local population varies from 0.03 microSv h(-1) to 0.83 microSv h(-1). The annual exposure dose to inhabitants in all the dwellings lies below the upper limit of 10 mSv given in ICRP-65.
Asunto(s)
Contaminación del Aire Interior/análisis , Exposición por Inhalación , Radón/análisis , Recolección de Datos , Monitoreo del Ambiente , Vivienda , Humanos , IndiaRESUMEN
Ion track filters (ITFs) are produced by physiochemical treatments to thin films of polymers and mica irradiated by heavy ions. These ITFs have many applications in the fields of science and technology. In the present investigation, the developed ITFs from polycarbonate films have been used to filter bacteria of various types in water. It is observed that the electric conduction through these filters depends upon the concentration of contaminants and pore diameter of filters. Filtration experiments were carried out using both single and multipore filters. Other applications related to environment surveillance have also been reported.
Asunto(s)
Monitoreo del Ambiente/instrumentación , Monitoreo del Ambiente/métodos , Microbiología del Agua , Bacterias , Filtración , Iones , PolímerosRESUMEN
A survey of indoor radon and thoron levels has been carried out in a number of villages in the vicinity of uranium bearing sites in the Hamirpur and Una districts of Himachal Pradesh (H.P.), India. Levels were analysed with reference to the nature of building material, soil type and different seasons of the year. The one year average for radon concentration was found to vary from a minimum of 19.7 to a maximum of 146.3 Bq/m3 while the minimum and maximum thoron concentrations were 9.1 and 70.7 Bq/m3, respectively. The dose rate varied from 0.1 to 8.67 microSv/h. These are discussed in the light of ICRP recommendations.