Characterization of transient and progressive pulmonary fibrosis by spatially correlated phase contrast microCT, classical histopathology and atomic force microscopy.

Pulmonary fibrosis (PF) is a severe and progressive condition in which the lung becomes scarred over time resulting in pulmonary function impairment. Classical histopathology remains an important tool for micro-structural tissue assessment in the diagnosis of PF. A novel workflow based on spatial correlated propagation-based phase-contrast micro computed tomography (PBI-microCT), atomic force microscopy (AFM) and histopathology was developed and applied to two different preclinical mouse models of PF - the commonly used and well characterized Bleomycin-induced PF and a novel mouse model for progressive PF caused by conditional Nedd4-2 KO. The aim was to integrate structural and mechanical features from hallmarks of fibrotic lung tissue remodeling. PBI-microCT was used to assess structural alteration in whole fixed and paraffin embedded lungs, allowing for identification of fibrotic foci within the 3D context of the entire organ and facilitating targeted microtome sectioning of planes of interest for subsequent histopathology. Subsequently, these sections of interest were subjected to AFM to assess changes in the local tissue stiffness of previously identified structures of interest. 3D whole organ analysis showed clear morphological differences in 3D tissue porosity between transient and progressive PF and control lungs. By integrating the results obtained from targeted AFM analysis, it was possible to discriminate between the Bleomycin model and the novel conditional Nedd4-2 KO model using agglomerative cluster analysis. As our workflow for 3D spatial correlation of PBI, targeted histopathology and subsequent AFM is tailored around the standard procedure of formalin-fixed paraffin-embedded (FFPE) tissue specimens, it may be a powerful tool for the comprehensive tissue assessment beyond the scope of PF and preclinical research.

The unique biomechanics of intermediate filaments - From single filaments to cells and tissues.

Together with actin filaments and microtubules, intermediate filaments (IFs) constitute the eukaryotic cytoskeleton and each of the three filament types contributes very distinct mechanical properties to this intracellular biopolymer network. IFs assemble hierarchically, rather than polymerizing from nuclei of a small number of monomers or dimers, as is the case with actin filaments and microtubules, respectively. This pathway leads to a molecular architecture specific to IFs and intriguing mechanical and dynamic properties: they are the most flexible cytoskeletal filaments and extremely extensible. Moreover, IFs are very stable against disassembly. Thus, they contribute important properties to cell mechanics, which recently have been investigated with state-of-the-art experimental and computational methods.

Membrane-Bound Vimentin Filaments Reorganize and Elongate under Strain.

Within a cell, intermediate filaments interact with other cytoskeletal components, altogether providing the cell's mechanical stability. However, little attention has been drawn to intermediate filaments close to the plasma membrane. In this cortex configuration, the filaments are coupled and arranged in parallel to the membrane, and the question arises of how they react to the mechanical stretching of the membrane. To address this question, we set out to establish an in vitro system composed of a polydimethylsiloxane-supported lipid bilayer. With a uniaxial stretching device, the supported membrane was stretched up to 34% in the presence of a lipid reservoir that was provided by adding small unilamellar vesicles in the solution. After vimentin attachment to the membrane, we observed structural changes of the vimentin filaments in networks of different densities by fluorescence microscopy and atomic force microscopy. We found that individual filaments respond to the membrane stretching with a reorganization along the stretching direction as well as an intrinsic elongation, while in a dense network, mainly filament reorganization was observed.

Life in plastic, it's not fantastic: Sublethal effects of polyethylene microplastics ingestion throughout amphibian metamorphosis.

Microplastics (MP) are an abundant, long-lasting, and widespread type of environmental pollution that is of increasing concern as it might pose a serious threat to ecosystems and species. However, these threats are still largely unknown for amphibians. Here, we used the African clawed frog (Xenopus laevis) as a model species to investigate whether polyethylene MP ingestion affects amphibian growth and development and leads to metabolic changes across two consecutive life stages (larvae and juveniles). Furthermore, we examined whether MP effects were more pronounced at higher rearing temperatures. Larval growth, development, and body condition were recorded, and standard metabolic rate (SMR) and levels of stress hormone (corticosterone, CORT) were measured. We determined variation in size, morphology, and hepatosomatic index in juveniles to identify any potential consequences of MP ingestion across metamorphosis. In both life stages, MP accumulation in the body was assessed. MP ingestion was found to result in sublethal effects on larval growth, development, and metabolism, to lead to allometric carry-over effects on juvenile morphology, and to accumulate in the specimens at both life stages. In larvae, SMR and developmental rate increased in response to MP ingestion; there additionally was a significant interaction of MP ingestion and temperature on development. CORT levels were higher in larvae that ingested MP, except at higher temperature. In juveniles, body was wider, and extremities were longer in animals exposed to MP during the larval stage; a high rearing temperature in combination with MP ingestion counteracted this effect. Our results provide first insights into the effects of MP on amphibians throughout metamorphosis and demonstrate that juvenile amphibians may act as a pathway for MP from freshwater to terrestrial environments. To allow for generalizations across amphibian species, future experiments need to consider the field prevalence and abundance of different MP in amphibians at various life stages.

Nefrectomía laparoscópica: experiencia clínica en 20 pacientes / Laparoscopic nephrectomy: clinical experience in 20 patients

La cirugía laparoscópica se está aplicando en una serie de patologías previamente sólo reservadas para la cirugía tradicional. Esta serie resume nuestra experiencia en 20 pacientes en los cuales realizamos una nefrectomía laparoscópica. Su margen de edad fue de 2 a 65 años, incluyendo 9 niños menores de 14 años. Efectuamos 16 nefrectomías simples, 2 nefrectomías parciales y 2 nefrectomías radicales por cáncer. El promedio de duración del procedimiento fue de 2,4 horas y la hospitalización de 2,5 días. Fue necesario convertir a cirugía abierta un caso por dificultad en la disección del pedículo renal y reoperamos una paciente por sangrado postoperatorio. No hubo mortalidad. La nefrectomía laparoscópica constituye un procedimiento quirúrgico alternativo efectivo