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Expert Thoughts about Therapeutic Climbing-A Multi-Perspective, Qualitative Examine.
Impaired hand function after neurological injuries can have a major impact on independence and quality of life. Most existing upper limb assessments are carried out in person, which is not always indicative of hand use in the community. Novel approaches to capture hand function in daily life are required to measure the true impact of rehabilitation interventions. Egocentric video combined with computer vision for automated analysis has been proposed to evaluate hand use at home. However, there are limitations to the duration of continuous recordings. We present a protocol designed to ensure that the videos obtained are representative of daily routines while respecting participant privacy. A representative recording schedule is selected through a collaborative process between the researchers and participants, to ensure that the videos capture natural tasks and performance, while being useful for hand assessment. Use of the equipment and procedures is demonstrated to the participants. A total of 3 h of video recordings are scheduled over two weeks. To reduce privacy concerns, participants have full control to start and stop recordings, and the opportunity to edit the videos before returning them to the research team. Reminders are provided, as well as help calls and home visits if necessary. The protocol was tested with 9 stroke survivors and 14 individuals with cervical spinal cord injury. The videos obtained contained a variety of activities, such as meal preparation, dishwashing, and knitting. An average of 3.11 ± 0.98 h of video were obtained. The recording periods varied from 12-69 d, due to illness or unexpected events in some cases. Data was successfully obtained from twenty-two out of 23 participants, with 6 participants requiring assistance from the investigators during the home recording period. The protocol was effective for collecting videos that contained valuable information about hand function at home after neurological injuries.Drosophila is an important model system to study a vast range of biological questions. Various organs and tissues from different developmental stages of the fly such as imaginal discs, the larval brain or egg chambers of adult females or the adult intestine can be extracted and kept in culture for imaging with time-lapse microscopy, providing valuable insights into cell and developmental biology. Here, we describe in detail our current protocol for the dissection of Drosophila larval brains, and then present our current approach for immobilizing and orienting larval brains and other tissues on a glass coverslip using Fibrin clots. This immobilization method only requires the addition of Fibrinogen and Thrombin to the culture medium. It is suitable for high-resolution time lapse imaging on inverted microscopes of multiple samples in the same culture dish, minimizes the lateral drifting frequently caused by movements of the microscope stage in multi-point visiting microscopy and allows for the addition and removal of reagents during the course of imaging. We also present custom-made macros that we routinely use to correct for drifting and to extract and process specific quantitative information from time-lapse analysis.Acoustically Targeted Chemogenetics (ATAC) allows for the noninvasive control of specific neural circuits. ATAC achieves such control through a combination of focused ultrasound (FUS) induced blood-brain barrier opening (FUS-BBBO), gene delivery with adeno-associated viral (AAV) vectors, and activation of cellular signaling with engineered, chemogenetic, protein receptors and their cognate ligands. With ATAC, it is possible to transduce both large and small brain regions with millimeter precision using a single noninvasive ultrasound application. This transduction can later allow for a long-term, noninvasive, device-free neuromodulation in freely moving animals using a drug. Since FUS-BBBO, AAVs, and chemogenetics have been used in multiple animals, ATAC should also be scalable for the use in other animal species. This paper expands upon a previously published protocol and outlines how to optimize the gene delivery with FUS-BBBO to small brain regions with MRI-guidance but without a need for a complicated MRI-compatible FUS device. The protocol, also, describes the design of mouse targeting and restraint components that can be 3D-printed by any lab and can be easily modified for different species or custom equipment. To aid reproducibility, the protocol describes in detail how the microbubbles, AAVs, and venipuncture were used in ATAC development. Finally, an example data is shown to guide the preliminary investigations of studies utilizing ATAC.When analyzing the activity of antimicrobial agents, it should be considered that microorganisms mainly occur in biofilms. Data obtained for planktonic bacteria cannot be transferred non-critically to biofilms. Biofilm models should consider both the relevant microorganisms and the conditions present in the environment. The selection of the model depends on the question to be answered. In dentistry, single species, multispecies, or microcosms originating from saliva or dental biofilm are used to culture biofilms. Microorganism selection depends on the focus of the study, for example caries biofilms mostly include Streptococcus mutans, an endodontic biofilm consists mostly of Enterococcus faecalis, and defined anaerobes are used in periodontal/peri-implant biofilms. In contrast to single-species biofilm models in medicine, where the lowest concentration of the antimicrobial that kills microorganisms is measured, the common analyzed variables are counts of colony-forming units or the percentage of dead bacteria determined by confocal laser scanning microscopy after applying a differentiating stain. All the models are helpful to evaluate new antimicrobial treatment options. Conclusions regarding the antimicrobial activity tendency of the therapeutics can be drawn. However, there are limitations of the model and ultimately a new therapy has to be proven in randomized controlled clinical trials.The intraoral biofilm requires mechanical removal due to its physical properties. When exposed to the biofilm, interdental areas need special devices to be used. The most effective ones and the first choice are interdental brushes. However, they may not be adequate in the case of very narrow interdental spaces. Despite the difficulties in handling, dental floss may have some advantage in subgingival cleaning. selleck chemical Data are present for gingivitis and periodontitis, but almost no information has been published on gingivally healthy individuals. With respect to interdental caries there is evidence that floss only has a protective value when used professionally and without sufficient fluoridation. There are no such data available on interdental brushes.
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