Bile duct ligation in the rat causes upregulation of ZO-2 and decreased colocalization of claudins with ZO-1 and occludin

As the only barrier between blood and bile compartments hepatocellular tight junctions play a crucial role in cholestasis-induced increase of biliary permeability. The molecular basis of this reversible defect is not known. We, therefore, examined expression, phosphorylation, distribution and colocalization of the junctional proteins occludin, claudin-1-3, ZO-1 and ZO-2 in rats after bile duct ligation and release of ligation. In control rats, claudin-1 and ZO-2 displayed a lobular gradient with highest expression levels in periportal cells, whereas claudin-2 showed a reciprocal distribution. Other proteins were evenly expressed in the liver lobule. Ligation resulted in upregulation of ZO-2 (2.7-fold), ZO-1 (1.4-fold) and occludin (1.2-fold) but not of claudins. Only ZO-2 showed increased phosphorylation. Distribution patterns were unchanged except for a strong accumulation of ZO-2 in perivenous hepatocytes. Colocalization analysis demonstrated that perivenous ZO-2 was the only protein examined revealing strongly increased overlap with occludin and ZO-1, whereas claudins and other proteins displayed a decrease. All changes were partially reversed by release of ligation. We conclude that differential expression of claudin-1-2 and ZO-2 has functional implications for bile formation. The moderately increased ZO-1 and occludin levels account for the known elongation of tight junction strands. The highly increased expression and changed distribution of ZO-2 suggests that ZO-1 is partly substituted by ZO-2, an alteration possibly causing impaired barrier functio

Epidermal Permeability Barrier: Transformation of Lamellar Granule-Disks into Intercellular Sheets by a Membrane-Fusion Process, a Freeze-Fracture Study

Freeze-fracture replication of lamellar granules and intercellular sheets of the horny layer in mouse, chicken, and snake epidermis reveals a pattern of serial fracture faces which is highly suggestive of polar lipids in a bilayer configuration. The occurrence of alternating wide and narrow fracture faces separated by intervening steps supports the view that epidermal barrier bilayers display lipid asymmetry similar to membranes. Within the lamellar granules, bilayers arrange to form disks which in fact are equivalent to flattened unilamellar liposomes. Stacking of the disks in turn gives rise to the lamellar pattern. After exocytosis into the intercellular space, the disks are arranged parallel to the cell membranes. In tangentially fractured specimens, the cleavage plane jumps back and forth from the plasma membrane to a disk-bilayer, thereby giving rise to the known phenomenon of EF-ridges (on the extracellular fracture face) and PE-grooves (in the plasmatic fracture face) which both represent the level of the plasma membrane sur- or subjacent to the aisles between disks. Concomitantly with the upward movement of the keratinocytes, the ridges and grooves become narrower until they fade away by the second or third cell layer of the stratum corneum. This phenomenon is explained by the fusion of adjacent disks at their highly curved brims due to a mechanism similar to the process of membrane fusion which causes the formation of wide, uninterrupted sheets

Stratum Corneum Lipid Liposomes: Calcium-Induced Transformation Into Lamellar Sheets

The epidermal water barrier in mammalian stratum corneum is formed of broad lamellar sheets of lipids consisting principally of ceramides (40%), cholesterol (25%), cholesterol sulfate (10%), and free fatty acids (25%). Such lipid mixtures have been shown to form lipid bilayers in the form of small, unilamellar liposomes when sonicated at 80°C in water containing Tris buffer and 100mM NaCl. In the present study it is shown that such liposomes are slowly transformed into large unilamellar liposomes and then into broad lamellar sheets after the addition of stoichiometric amounts of calcium chloride. The presence of free fatty acids was a necessary condition for this calcium-induced fusion. These observations may provide a useful analogy for the transformation of flattened liposomes into broad lamellar sheets that occurs during transition of epidermal granular cells into corneocytes

Large-scale laboratory experiments on mussel dropper lines in ocean surface waves

The rapid growth of marine aquaculture around the world accentuates issues of sustainabil-ity and environmental impacts of large-scale farming systems. One potential mitigation strategy is to relocate to more energetic offshore locations. However, research regarding the forces which waves and currents impose on aquaculture structures in such conditions is still scarce. The present study aimed at extending the knowledge related to live blue mussels (Mytilus edulis), cultivated on dropper lines, by unique, large-scale laboratory experiments in the Large Wave Flume of the Coastal Research Center in Hannover, Germany. Nine-months-old live dropper lines and a surrogate of 2.0 m length each are exposed to regular waves with wave heights between 0.2 and 1.0 m and periods between 1.5 and 8.0 s. Force time histories are recorded to investigate the inertia and drag characteristics of live mussel and surrogate dropper lines. The surrogate dropper line was developed from 3D scans of blue mussel dropper lines, using the surface descriptor Abbott–Firestone Curve as quality parameter. Pull-off tests of individual mussels are conducted that reveal maximum attachment strength ranges of 0.48 to 10.55 N for mussels that had medium 3.04 cm length, 1.60 cm height and 1.25 cm width. Mean drag coefficients of CD = 3.9 were found for live blue mussel lines and CD = 3.4 for the surrogate model, for conditions of Keulegan–Carpenter number (KC) 10 to 380, using regular wave tests. © 2020 by the authors. Licensee MDPI, Basel, Switzerland

The CD44 standard/ezrin complex regulates Fas-mediated apoptosis in Jurkat cells

The transmembrane receptor CD44 conveys important signals from the extracellular microenvironment to the cytoplasm, a phenomena known as "outside-in” signaling. CD44 exists as several isoforms that result from alternative splicing, which differ only in the extracellular domain but yet exhibit different activities. CD44 is a binding partner for the membrane-cytoskeleton cross-linker protein ezrin. In this study, we demonstrate that only CD44 standard (CD44s) colocalizes and interacts with the actin cross-linkers ezrin and moesin using well-characterized cell lines engineered to express different CD44 isoforms. Importantly, we also show that the association CD44s-ezrin-actin is an important modulator of Fas-mediated apoptosis. The results highlight a mechanism by which signals from the extracellular milieu regulate intracellular signaling activities involved in programmed cell deat

A new system design for the cultivation of extractive species at exposed sites - Part 2: Experimental modelling in waves and currents

Aquaculture is projected to be a major supplier of marine proteins to large parts of the global population. This includes bivalves, which have a high potential to offset protein deficits, as they are highly adaptable to varying water temperature, salinity, desiccation, and oxygen conditions. This work is part of a two-piece contribution on novel marine aquaculture technology and details physical laboratory tests of a new cultivation system for bivalve farming called “Shellfish Tower”. The tested 1:20 model consists of a rectangular cage (2 × 2 m prototype scale) with a central buoyancy element and a height of 2 – 4 m. Testing was done in a current flume as well as a wave basin for current velocities between 0.4 – 2.2 m/s and wave heights of 1.6 to 5.0 m with periods between 5 to 14 s. The tests were conducted to prove the feasibility and functionality of this aquaculture system, which is usable for the collection and cultivation of mussel spat as well as for the grow-out of oysters, scallops, and seaweed in marine environments. Tests carried out in a current flume revealed that drag coefficients decrease with increasing current velocities, and range from Cd=0.5 to 2.5, while the mooring inclination increases from 12° to 84° with increasing flow velocity, which is highly dependant on the buoyancy related pretension. The examination of the mooring line tensions recorded in a wave basin showed that the largest values of snap-induced tension were up to 10 times that of the semi-static tension. The maximum-recorded tension on the system was 48 kN for a single and 89 kN for a double configuration, compared to non-snap tension values, which were in the range of 6 – 10 kN. The insights gathered in this study will inform the future design of aquaculture systems in high-energy environments and allow for an integration into numerical models

Epitope-engineered human hematopoietic stem cells are shielded from CD123-targeted immunotherapy

Targeted eradication of transformed or otherwise dysregulated cells using monoclonal antibodies (mAb), antibody-drug conjugates (ADC), T cell engagers (TCE), or chimeric antigen receptor (CAR) cells is very effective for hematologic diseases. Unlike the breakthrough progress achieved for B cell malignancies, there is a pressing need to find suitable antigens for myeloid malignancies. CD123, the interleukin-3 (IL-3) receptor alpha-chain, is highly expressed in various hematological malignancies, including acute myeloid leukemia (AML). However, shared CD123 expression on healthy hematopoietic stem and progenitor cells (HSPCs) bears the risk for myelotoxicity. We demonstrate that epitope-engineered HSPCs were shielded from CD123-targeted immunotherapy but remained functional, while CD123-deficient HSPCs displayed a competitive disadvantage. Transplantation of genome-edited HSPCs could enable tumor-selective targeted immunotherapy while rebuilding a fully functional hematopoietic system. We envision that this approach is broadly applicable to other targets and cells, could render hitherto undruggable targets accessible to immunotherapy, and will allow continued posttransplant therapy, for instance, to treat minimal residual disease (MRD)

Large-Scale Laboratory Experiments on Mussel Dropper Lines in Ocean Surface Waves

The rapid growth of marine aquaculture around the world accentuates issues of sustainability and environmental impacts of large-scale farming systems. One potential mitigation strategy is to relocate to more energetic offshore locations. However, research regarding the forces which waves and currents impose on aquaculture structures in such conditions is still scarce. The present study aimed at extending the knowledge related to live blue mussels (Mytilus edulis), cultivated on dropper lines, by unique, large-scale laboratory experiments in the Large Wave Flume of the Coastal Research Center in Hannover, Germany. Nine-months-old live dropper lines and a surrogate of 2.0 m length each are exposed to regular waves with wave heights between 0.2 and 1.0 m and periods between 1.5 and 8.0 s. Force time histories are recorded to investigate the inertia and drag characteristics of live mussel and surrogate dropper lines. The surrogate dropper line was developed from 3D scans of blue mussel dropper lines, using the surface descriptor Abbott–Firestone Curve as quality parameter. Pull-off tests of individual mussels are conducted that reveal maximum attachment strength ranges of 0.48 to 10.55 N for mussels that had medium 3.04 cm length, 1.60 cm height and 1.25 cm width. Mean drag coefficients of CD = 3.9 were found for live blue mussel lines and CD = 3.4 for the surrogate model, for conditions of Keulegan–Carpenter number (KC) 10 to 380, using regular wave tests

The Magic of Stonehenge or How to Predict Solar and Lunar Eclipses

In this paper, the authors present a self-developed app that features the Stonehenge algorithm for predicting lunar and solar eclipses. Furthermore, the app presents useful guidelines for astronomy photography and enhances optics and photonics training