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- Klionsky, Daniel J., et al.
(författare)
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Guidelines for the use and interpretation of assays for monitoring autophagy
- 2012
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Ingår i: Autophagy. - : Informa UK Limited. - 1554-8635 .- 1554-8627. ; 8:4, s. 445-544
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Forskningsöversikt (refereegranskat)abstract
- In 2008 we published the first set of guidelines for standardizing research in autophagy. Since then, research on this topic has continued to accelerate, and many new scientists have entered the field. Our knowledge base and relevant new technologies have also been expanding. Accordingly, it is important to update these guidelines for monitoring autophagy in different organisms. Various reviews have described the range of assays that have been used for this purpose. Nevertheless, there continues to be confusion regarding acceptable methods to measure autophagy, especially in multicellular eukaryotes. A key point that needs to be emphasized is that there is a difference between measurements that monitor the numbers or volume of autophagic elements (e.g., autophagosomes or autolysosomes) at any stage of the autophagic process vs. those that measure flux through the autophagy pathway (i.e., the complete process); thus, a block in macroautophagy that results in autophagosome accumulation needs to be differentiated from stimuli that result in increased autophagic activity, defined as increased autophagy induction coupled with increased delivery to, and degradation within, lysosomes (in most higher eukaryotes and some protists such as Dictyostelium) or the vacuole (in plants and fungi). In other words, it is especially important that investigators new to the field understand that the appearance of more autophagosomes does not necessarily equate with more autophagy. In fact, in many cases, autophagosomes accumulate because of a block in trafficking to lysosomes without a concomitant change in autophagosome biogenesis, whereas an increase in autolysosomes may reflect a reduction in degradative activity. Here, we present a set of guidelines for the selection and interpretation of methods for use by investigators who aim to examine macroautophagy and related processes, as well as for reviewers who need to provide realistic and reasonable critiques of papers that are focused on these processes. These guidelines are not meant to be a formulaic set of rules, because the appropriate assays depend in part on the question being asked and the system being used. In addition, we emphasize that no individual assay is guaranteed to be the most appropriate one in every situation, and we strongly recommend the use of multiple assays to monitor autophagy. In these guidelines, we consider these various methods of assessing autophagy and what information can, or cannot, be obtained from them. Finally, by discussing the merits and limits of particular autophagy assays, we hope to encourage technical innovation in the field.
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| 2. |
- Carmona-Gutierrez, D., et al.
(författare)
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Guidelines and recommendations on yeast cell death nomenclature
- 2018
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Ingår i: Microbial Cell. - : Shared Science Publishers OG. - 2311-2638. ; 5:1, s. 4-31
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Forskningsöversikt (refereegranskat)abstract
- Elucidating the biology of yeast in its full complexity has major implications for science, medicine and industry. One of the most critical processes determining yeast life and physiology is cellular demise. However, the investigation of yeast cell death is a relatively young field, and a widely accepted set of concepts and terms is still missing. Here, we propose unified criteria for the definition of accidental, regulated, and programmed forms of cell death in yeast based on a series of morphological and biochemical criteria. Specifically, we provide consensus guidelines on the differential definition of terms including apoptosis, regulated necrosis, and autophagic cell death, as we refer to additional cell death routines that are relevant for the biology of (at least some species of) yeast. As this area of investigation advances rapidly, changes and extensions to this set of recommendations will be implemented in the years to come. Nonetheless, we strongly encourage the authors, reviewers and editors of scientific articles to adopt these collective standards in order to establish an accurate framework for yeast cell death research and, ultimately, to accelerate the progress of this vibrant field of research.
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| 3. |
- Kucharíková, S., et al.
(författare)
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Covalent immobilization of antimicrobial agents on titanium prevents Staphylococcus aureus and Candida albicans colonization and biofilm formation
- 2016
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Ingår i: Journal of Antimicrobial Chemotherapy. - : Oxford University Press. - 0305-7453 .- 1460-2091. ; 71:4, s. 936-945
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Tidskriftsartikel (refereegranskat)abstract
- Objectives: Biofilm-associated implant infections represent a serious public health problem. Covalent immobilization of antimicrobial agents on titanium (Ti), thereby inhibiting biofilm formation of microbial pathogens, is a solution to this problem. Methods: Vancomycin (VAN) and caspofungin (CAS) were covalently bound on Ti substrates using an improved processing technique adapted to large-scale coating of implants. Resistance of the VAN-coated Ti (VAN-Ti) and CAS-coated Ti (CAS-Ti) substrates against in vitro biofilm formation of the bacterium Staphylococcus aureus and the fungal pathogen Candida albicans was determined by plate counting and visualized by confocal laser scanning microscopy. The efficacy of the coated Ti substrates was also tested in vivo using an adapted biomaterial-associated murine infection model in which control-Ti, VAN-Ti or CAS-Ti substrates were implanted subcutaneously and subsequently challenged with the respective pathogens. The osseointegration potential of VAN-Ti and CAS-Ti was examined in vitro using human bone marrow-derived stromal cells, and for VAN-Ti also in a rat osseointegration model. Results: In vitro biofilm formation of S. aureus and C. albicans on VAN-Ti and CAS-Ti substrates, respectively, was significantly reduced compared with biofilm formation on control-Ti. In vivo, we observed over 99.9% reduction in biofilm formation of S. aureus on VAN-Ti substrates and 89% reduction in biofilm formation of C. albicans on CAS-Ti substrates, compared with control-Ti substrates. The coated substrates supported osseointegration in vitro and in vivo. Conclusions: These data demonstrate the clinical potential of covalently bound VAN and CAS on Ti to reduce microbial biofilm formation without jeopardizing osseointegration.
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| 8. |
- Thevissen, E, et al.
(författare)
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The provision of oral hygiene instructions and patient motivation in a dental care system without dental hygienists
- 2017
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Ingår i: International Journal of Dental Hygiene. - : John Wiley & Sons. - 1601-5029 .- 1601-5037. ; 15:4, s. 261-268
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Tidskriftsartikel (refereegranskat)abstract
- INTRODUCTION: Oral hygiene instructions (OHI) and patient motivation (PM), often provided by dental hygienists, are essential attributes to establishing good oral health in patients. The purpose of this study was to investigate the provision of OHI and PM by dental professionals in a dental care system without dental hygienists. METHODS AND MATERIALS: A questionnaire, including 22 items (demographics, OHI and PM), was presented to 1037 dental professionals (=21.6% Flemish population). Descriptive statistics were used to analyse the provision of OHI and PM. Additionally, the Fisher-Freeman-Halton exact test was used to investigate the influence of qualification (general practitioner/specialist), work experience (< or >/= 25 years), workload (< or >/= 30 h), location of practice (rural/urban) and chair-side assistance (with/without). RESULTS: Response rate was 75%. Participants reported a single strategy for preventive care and felt their efforts were not in harmony with the results. Lack of time, remuneration and patient interest were reported as complicating factors and participants agreed on allowing assistants to provide preventive care. Significant variance was found based on qualification (12/17 items), work experience (7/17 items), workload (1/17 items), location of practice (2/17 items) and chair-side assistance (15/17 items). CONCLUSION: In a context without dental hygienists, OHI and PM appeared non-compliant with current international guidelines. Although dental professionals were concerned with preventive dentistry, they reported barriers including lack of time, remuneration and patient compliance. Almost all participants expressed the need for delegation of in-mouth OHI, suggesting a need for dental hygienists.
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| 9. |
- Vandeweghe, Stefan, et al.
(författare)
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Immediate loading of screw-retained all-ceramic crowns in immediate versus delayed single implant placement
- 2013
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Ingår i: International Journal of Prosthodontics. - : Quintessence. - 0893-2174 .- 1139-9791. ; 26:5, s. 458-464
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Tidskriftsartikel (refereegranskat)abstract
- PURPOSE: The aim of this study was to evaluate and compare the outcomes of immediately loaded all-ceramic crown restorations supported by implants placed in both mature bone and immediate extraction sockets. MATERIALS AND METHODS: Forty-three tapered, external hex implants were placed immediately after extraction or in healed bone in a convenience sample of 38 patients from two clinics, with 23 implants immediately placed after tooth extraction and 20 placed in mature bone. Thirty implants were located in the maxilla and 13 in the mandible. Each surgical implant placement was followed by the fabrication of a screw-retained all-ceramic crown (zirconia-toughened alumina cylinder and layered with porcelain) that was then put into immediate function. Each patient was recalled annually for clinical and radiographic assessments. RESULTS: All implants were loaded immediately and all functioned successfully following a mean follow-up of 26 months. Significantly more bone loss was recorded around delayed implants compared with immediately placed ones. Smoking, platform switching, and jaw location did not influence peri-implant bone loss. Porcelain chipping was noted in the case of two crowns. CONCLUSION: The short-term outcome of prefabricated ceramic cylinders used to support a single screw-retained crown was regarded as successful. Immediate implant placement did not appear to increase the risk for implant failure and was accompanied by reduced peri-implant bone loss.
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