In 2008, we published the first set of guidelines for standardizing research in autophagy. Since then, this topic has received increasing attention, and many scientists have entered the field. Our knowledge base and relevant new technologies have also been expanding. Thus, it is important to formulate on a regular basis updated guidelines for monitoring autophagy in different organisms. Despite numerous reviews, there continues to be confusion regarding acceptable methods to evaluate autophagy, especially in multicellular eukaryotes. Here, we present a set of guidelines for investigators to select and interpret methods to examine autophagy and related processes, and for reviewers to provide realistic and reasonable critiques of reports that are focused on these processes. These guidelines are not meant to be a dogmatic set of rules, because the appropriateness of any assay largely depends on the question being asked and the system being used. Moreover, no individual assay is perfect for every situation, calling for the use of multiple techniques to properly monitor autophagy in each experimental setting. Finally, several core components of the autophagy machinery have been implicated in distinct autophagic processes (canonical and noncanonical autophagy), implying that genetic approaches to block autophagy should rely on targeting two or more autophagy-related genes that ideally participate in distinct steps of the pathway. Along similar lines, because multiple proteins involved in autophagy also regulate other cellular pathways including apoptosis, not all of them can be used as a specific marker for bona fide autophagic responses. Here, we critically discuss current methods of assessing autophagy and the information they can, or cannot, provide. Our ultimate goal is to encourage intellectual and technical innovation in the field.
- MeSH
- Autophagy * physiology MeSH
- Autophagosomes MeSH
- Biomarkers MeSH
- Biological Assay standards MeSH
- Humans MeSH
- Lysosomes MeSH
- Autophagy-Related Proteins metabolism MeSH
- Animals MeSH
- Check Tag
- Humans MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Research Support, N.I.H., Extramural MeSH
- Research Support, U.S. Gov't, Non-P.H.S. MeSH
- Guideline MeSH
Importance: Early diagnosis of retinoblastoma, the most common intraocular cancer, can save both a child's life and vision. However, anecdotal evidence suggests that many children across the world are diagnosed late. To our knowledge, the clinical presentation of retinoblastoma has never been assessed on a global scale. Objectives: To report the retinoblastoma stage at diagnosis in patients across the world during a single year, to investigate associations between clinical variables and national income level, and to investigate risk factors for advanced disease at diagnosis. Design, Setting, and Participants: A total of 278 retinoblastoma treatment centers were recruited from June 2017 through December 2018 to participate in a cross-sectional analysis of treatment-naive patients with retinoblastoma who were diagnosed in 2017. Main Outcomes and Measures: Age at presentation, proportion of familial history of retinoblastoma, and tumor stage and metastasis. Results: The cohort included 4351 new patients from 153 countries; the median age at diagnosis was 30.5 (interquartile range, 18.3-45.9) months, and 1976 patients (45.4%) were female. Most patients (n = 3685 [84.7%]) were from low- and middle-income countries (LMICs). Globally, the most common indication for referral was leukocoria (n = 2638 [62.8%]), followed by strabismus (n = 429 [10.2%]) and proptosis (n = 309 [7.4%]). Patients from high-income countries (HICs) were diagnosed at a median age of 14.1 months, with 656 of 666 (98.5%) patients having intraocular retinoblastoma and 2 (0.3%) having metastasis. Patients from low-income countries were diagnosed at a median age of 30.5 months, with 256 of 521 (49.1%) having extraocular retinoblastoma and 94 of 498 (18.9%) having metastasis. Lower national income level was associated with older presentation age, higher proportion of locally advanced disease and distant metastasis, and smaller proportion of familial history of retinoblastoma. Advanced disease at diagnosis was more common in LMICs even after adjusting for age (odds ratio for low-income countries vs upper-middle-income countries and HICs, 17.92 [95% CI, 12.94-24.80], and for lower-middle-income countries vs upper-middle-income countries and HICs, 5.74 [95% CI, 4.30-7.68]). Conclusions and Relevance: This study is estimated to have included more than half of all new retinoblastoma cases worldwide in 2017. Children from LMICs, where the main global retinoblastoma burden lies, presented at an older age with more advanced disease and demonstrated a smaller proportion of familial history of retinoblastoma, likely because many do not reach a childbearing age. Given that retinoblastoma is curable, these data are concerning and mandate intervention at national and international levels. Further studies are needed to investigate factors, other than age at presentation, that may be associated with advanced disease in LMICs.
- MeSH
- Infant MeSH
- Humans MeSH
- Child, Preschool MeSH
- Retinoblastoma economics epidemiology MeSH
- Check Tag
- Infant MeSH
- Humans MeSH
- Male MeSH
- Child, Preschool MeSH
- Female MeSH
- Publication type
- Journal Article MeSH
The current research confirmed the presence of mi-croplastics in the environment and also in living organ-isms. Collection, separation and identification of micro-plastics from environment are very desirable activities, important for monitoring these undesirable materials. In order to determine the real risk brought about by plastic fragments, it is necessary to implement a standard protocol for their sampling, quantification and identification. In this study, analytical techniques, such as SEM, py-GC-MS, FTIR, NMR and Raman spectroscopy, often used for the identification of microplastics, are discussed, together with novel methods used to identify the pollutants adsorbed on them, e.g., pharmaceuticals and metals.
Polyurethane has a good insulation characteristic, and it is widely used as an insulation and lining material for refrigerators. Nevertheless, at the end of a product's lifetime, serious problems arise related to waste management. Recently, energy recovery has been marked as a promising solution, especially, waste-to-energy applications. To find an appropriate application for such waste, a thermal analysis was performed. An experimental analysis of polyurethane (PU) waste material was performed by the method of pyrolysis gas chromatography with mass spectrometric detection (Py-GC/MS) at various pyrolytic temperatures, namely, at 500, 600 and 700 °C. Waste polyurethane foam was conducted to investigations in the form of a bulk sample and sorted grain-size samples with a goal to detect the chemical composition of the pyrolysate. The investigation revealed various groups of organic compounds such as heterocyclic compounds of nitrogen and simple and polycyclic aromatic hydrocarbons, while notable concentrations of compounds containing chlorine were detected as well. The experimental analysis found differences in the composition of amines and other compounds and in the dependence on grain size composition. Bulk samples produced the highest concentration of amines (˃ 40%) at a temperature of 500 °C. The sample homogenization led to a significant increase in amines production. Polyurethane waste of grain size from 0.125 to 0.25 mm contained approximately 80% amines. The pyrolysis of the bulk sample at 600 °C yielded only 24% nitrogen heterocyclic compounds, while from the sorted grain-size <0.045 mm and 0.045-0.063 mm, yields were increased to 47.4 and 45.7%, respectively.
- MeSH
- Autophagy * physiology MeSH
- Biological Assay methods standards MeSH
- Humans MeSH
- Computer Simulation MeSH
- Animals MeSH
- Check Tag
- Humans MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
- Research Support, N.I.H., Extramural MeSH
- Guideline MeSH
Contents of uranium in coals from Odeř in the northernmost part of the Sokolov Basin, Czech Republic, in the vicinity of the well known St. Joachimsthal uranium ore deposits, reach extremely high values. In the present work, coal samples with contents of uranium ranging from 0.02 to 6 wt.% were studied. The study employing a whole complex of analytical techniques has been aimed at identification of changes in the structure of coal organic matter, which are associated with the high contents of uranium in coal. The study includes proximate and ultimate analyses, multielement analysis by instrumental neutron and photon activation analyses, micropetrographic analysis by optical microscopy, ESEM/EDX analysis of mineral matter, infrared and Raman spectroscopies, solvent extraction followed by gas chromatography with mass spectroscopy (GC/MS), and analytical pyrolysis (Py-GC/MS). The study has confirmed previously proposed explanation of uraniferous mineralization in sedimentary carboniferous substances by the mechanism of reduction and fixation of soluble U(VI) (uranyl, UO2(2+)) species (e.g., humic, carbonate/hydroxo/phosphate complexes) by sedimentary organic matter under diagenetic or hydrothermal conditions, and formation of insoluble U(IV) species as phosphate minerals and uraninite. The process is accompanied with alteration and destruction of the coal organic matter. The changes in the structure of coal organic matter involve dehydrogenation and oxidation mainly in the aliphatic, aromatic and hydroxyl structures, and an increase in aromaticity, content of ether bonds, and the degree of coalification.
- MeSH
- Radioactive Pollutants analysis MeSH
- Coal analysis MeSH
- Uranium analysis MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Geographicals
- Czech Republic MeSH
