The OPFOS microscopy family: High-resolution optical-sectioning of biomedical specimens

We report on the recently emerging (Laser) Light Sheet based Fluorescence Microscopy field (LSFM). The techniques used in this field allow to study and visualize biomedical objects non-destructively in high-resolution through virtual optical sectioning with sheets of laser light. Fluorescence originating in the cross section of the sheet and sample is recorded orthogonally with a camera. In this paper, the first implementation of LSFM to image biomedical tissue in three dimensions - Orthogonal-Plane Fluorescence Optical Sectioning microscopy (OPFOS) - is discussed. Since then many similar and derived methods have surfaced (SPIM, Ultramicroscopy, HR-OPFOS, mSPIM, DSLM, TSLIM...) which we all briefly discuss. All these optical sectioning methods create images showing histological detail. We illustrate the applicability of LSFM on several specimen types with application in biomedical and life sciences.Comment: 19 pages, 10 figures, to be published in Anatomical Research International (Hindawi

Seasonal water storage and release dynamics of bofedal wetlands in the Central Andes

Tropical high-Andean wetlands, locally known as ‘bofedales’, are key ecosystems sustaining biodiversity, carbon sequestration, water provision and livestock farming. Bofedales' contribution to dry season baseflows and sustaining water quality is crucial for downstream water security. The sensitivity of bofedales to climatic and anthropogenic disturbances is therefore of growing concern for watershed management. This study aims to understand seasonal water storage and release characteristics of bofedales by combining remote sensing analysis and ground-based monitoring for the wet and dry seasons of late 2019 to early 2021, using the glacierised Vilcanota-Urubamba basin (Southern Peru) as a case study. A network of five ultrasound loggers was installed to obtain discharge and water table data from bofedal sites across two headwater catchments. The seasonal extent of bofedales was mapped by applying a supervised machine learning model using Random Forest on imagery from Sentinel-2 and NASADEM. We identified high seasonal variability in bofedal area with a total of 3.5% and 10.6% of each catchment area, respectively, at the end of the dry season (2020), which increased to 15.1% and 16.9%, respectively, at the end of the following wet season (2021). The hydrological observations and bofedal maps were combined into a hydrological conceptual model to estimate the storage and release characteristics of the bofedales, and their contribution to runoff at the catchment scale. Estimated lag times between 1 and 32 days indicate a prolonged bofedal flow contribution throughout the dry season (about 74% of total flow). Thus, our results suggest that bofedales provide substantial contribution to dry season baseflow, water flow regulation and storage. These findings highlight the importance of including bofedales in local water management strategies and adaptation interventions including nature-based solutions that seek to support long-term water security in seasonally dry and rapidly changing Andean catchments

Molecular mechanisms of cell death: recommendations of the Nomenclature Committee on Cell Death 2018.

Over the past decade, the Nomenclature Committee on Cell Death (NCCD) has formulated guidelines for the definition and interpretation of cell death from morphological, biochemical, and functional perspectives. Since the field continues to expand and novel mechanisms that orchestrate multiple cell death pathways are unveiled, we propose an updated classification of cell death subroutines focusing on mechanistic and essential (as opposed to correlative and dispensable) aspects of the process. As we provide molecularly oriented definitions of terms including intrinsic apoptosis, extrinsic apoptosis, mitochondrial permeability transition (MPT)-driven necrosis, necroptosis, ferroptosis, pyroptosis, parthanatos, entotic cell death, NETotic cell death, lysosome-dependent cell death, autophagy-dependent cell death, immunogenic cell death, cellular senescence, and mitotic catastrophe, we discuss the utility of neologisms that refer to highly specialized instances of these processes. The mission of the NCCD is to provide a widely accepted nomenclature on cell death in support of the continued development of the field

Quasi-static Transfer Function of the Rabbit Middle Ear‚ Measured with a Heterodyne Interferometer with High-Resolution Position Decoder

Due to changes in ambient pressure and to the gas-exchange processes in the middle ear (ME) cavity, the ear is subject to ultra-low-frequency pressure variations, which are many orders of magnitude larger than the loudest acoustic pressures. Little quantitative data exist on how ME mechanics deals with these large quasi-static pressure changes and because of this lack of data, only few efforts could be made to incorporate quasi-static behavior into computer models. When designing and modeling ossicle prostheses and implantable ME hearing aids, the effects of large ossicle movements caused by quasi-static pressures should be taken into account. We investigated the response of the ME to slowly varying pressures by measuring the displacement of the umbo and the stapes in rabbit with a heterodyne interferometer with position decoder. Displacement versus pressure curves were obtained at linear pressure change rates between 200 Pa/s and 1.5 kPa/s, with amplitude ±2.5 kPa. The change in stapes position associated with a pressure change is independent of pressure change rate (34 μm peak-to-peak at ±2.5 kPa). The stapes displacement versus pressure curves are highly nonlinear and level off for pressures beyond ±1 kPa. Stapes motion shows no measurable hysteresis at 1.5 kPa/s, which demonstrates that the annular ligament has little viscoelasticity. Hysteresis increases strongly at the lowest pressure change rates. The stapes moves in phase with the umbo and with pressure, but the sense of rotation of the hysteresis loop of stapes is phase inversed. Stapes motion is not a simple lever ratio mimic of umbo motion, but is the consequence of complex changes in ossicle joints and ossicle position. The change in umbo position produced by a ±2.5 kPa pressure change decreases with increasing rate from 165 μm at 200 Pa/s to 118 μm at 1.5 kPa/s. Umbo motion already shows significant hysteresis at 1.5 kPa/s, but hysteresis increases further as pressure change rate decreases. We conclude that in the quasi-static regime, ossicle movement is not only governed by viscoelasticity, but that other effects become dominant as pressure change rate decreases below 1 kPa/s. The increasing hysteresis can be caused by increasing friction as speed of movement decreases, and incorporating speed-dependent friction coefficients will be essential to generate realistic models of ossicle movements at slow pressure change rates

mTHPC-mediated Photodynamic Therapy of Early Stage Oral Squamous Cell Carcinoma:A Comparison to Surgical Treatment

<p>mTHPC-mediated photodynamic therapy (PDT) is used for treatment of early head and neck squamous cell carcinoma. This study is a retrospective comparison of PDT with transoral surgery in the treatment of early primary squamous cell carcinoma of the oral cavity/oropharynx.</p><p>PDT data were retrieved from four study databases; surgical results were retrieved from our institutional database. To select similar primary tumors, infiltration depth was restricted to 5 mm for the surgery group. A total of 126 T1 and 30 T2 tumors were included in the PDT group, and 58 T1 and 33 T2 tumors were included in the surgically treated group.</p><p>Complete response rates with PDT and surgery were 86 and 76 % for T1, respectively, and for T2 63 and 78 %. Lower local disease-free survival for PDT compared to surgery was found. However, when comparing the need for local retreatment, no significant difference for T1 tumors was found, while for T2 tumors surgery resulted in significantly less need for local retreatment. No significant differences in overall survival between surgery and PDT were observed.</p><p>PDT for T1 tumors results in a similar need for retreatment compared to surgery, while for T2 tumors PDT performs worse. Local disease-free survival for surgery is better than for PDT. This may be influenced by the benefit surgery has of having histology available. This allows an early decision on reintervention, while for PDT one has to follow a wait-and-see policy. Future prospective studies should compare efficacy as well as morbidity.</p>