Search

This work presents a compact plant's development and performance evaluation for the decentralized treatment of domestic sewage. The plant was conceived and installed in a house with four residents in Vicente Pires, Federal District, Brazil. Its purpose was to remove organic matter and solids using a low-cost biological treatment process that was simple to operate. The plant was essentially anaerobic, composed of an up-flow anaerobic reactor and an anaerobic filter, both associated with lamella settlers. It was operated under real conditions and monitored for nineteen months, with removal efficiencies (calculated over the medians) of 81% for COD, 83% for BOD, 51% for Total Solids, 55% for Total Volatile Solids, 87% for Total Suspended Solids, and 100% for Settleable Solids. The plant performed adequately, with no clogging between the plates of the lamella settlers, no offensive odours, and limited amounts of sludge and scum.

Introduction: The anterosuperior portion of the bulla lamella can extend into the frontal sinus and form the supra bulla frontal cell (SBFC) and supraorbital ethmoid cell (SOEC). This can affect the frontal drainage pathway and make surgery more challenging. Objectives: The aim of the study was to compare the anatomical characteristics of SBFC and SOEC in Chinese chronic rhinosinusitis (CRS) patients and explore the relationship between the cells and frontal sinusitis (FS). The surgical skills in dealing with these cells were also studied. Methods: We prospectively identified SBFC and SOEC in 114 patients with CRS by computed tomography (CT). The sides of the patients were divided into groups A (with FS) and B (without FS). CT scans were analyzed to distinguish the SBFC, SOEC, and the drainage pathway. Statistical analysis was conducted to determine whether the cells were associated with the occurrence of FS. Results: The prevalence of frontal cells was as follows: SBFC: 9.65%, SOEC: 21.93%. There was an association between the presence of SBFC and FS (p < 0.05). The anterior drainage pathway was present in patients with SBFC and SOEC, anterolateral pathway in those with SBFC/SOEC coexisting with the frontal septal cell, and anteromedial pathway in those with SBFC/SOEC coexisting with the supra agger frontal cell. Conclusions: There was an association between the presence of SBFC and the occurrence of FS. Extension of the anterosuperior portion of the bulla lamella into the frontal sinus obstructed the drainage pathway posteriorly. Understanding the anatomy may help surgeons thoroughly dissect the bulla lamella.

We reported a case of upper eyelid necrosis initially misdiagnosed as a preseptal cellulitis following a hordeolum externum resulting in great damage to the upper eyelid (anterior lamella). The infection was successfully treated with surgical cleansing, drainage, and endovenous antibiotics. Early treatment may avoid severe complications such as eyelid deformity, systemic involvement, and blindness.

Lamellar bodies (LBs) are surfactant-rich organelles in alveolar cells. LBs disassemble into a lipid-protein network that reduces surface tension and facilitates gas exchange in the alveolar cavity. Current knowledge of LB architecture is predominantly based on electron microscopy studies using disruptive sample preparation methods. We established and validated a post-correlation on-lamella cryo-correlative light and electron microscopy approach for cryo-FIB milled cells to structurally characterize and validate the identity of LBs in their unperturbed state. Using deconvolution and 3D image registration, we were able to identify fluorescently labeled membrane structures analyzed by cryo-electron tomography. In situ cryo-electron tomography of A549 cells as well as primary Human Small Airway Epithelial Cells revealed that LBs are composed of membrane sheets frequently attached to the limiting membrane through “T”-junctions. We report a so far undescribed outer membrane dome protein complex (OMDP) on the limiting membrane of LBs. Our data suggest that LB biogenesis is driven by parallel membrane sheet import and by the curvature of the limiting membrane to maximize lipid storage capacity., Using the post-correlation on-lamella cryo-CLEM workflow, Klein, Wimmer et al. show that lamellar bodies (LBs) are composed of membrane sheets frequently attached to the limiting membrane through T-junctions in ABCA3 overexpressing cells and in primary human small airway epithelial cells. This study provides insights into LB biogenesis and membrane packing inside the LB.

We present a case of a 75-year-old patient who underwent reconstruction of the lower eyelid and lateral canthus following removal of 80% of the lower eyelid due to basal cell carcinoma. A Hughes transconjunctival eyelid-sharing flap was used to form the posterior lamella, and a free skin graft from the ipsilateral upper lid was used to recreate the anterior lamella. The lateral canthal ligament was reconstructed using a free fascia lata graft. A periosteal flap was not used due to local scarring that was the result of previous multiple lateral canthal surgeries. Skin preservation in the lateral canthal area and additional horizontal support for the lower eyelid were achieved by using a local advancement flap. This two-stage surgery produced excellent functional and cosmetic effects. Fascia lata free graft can be an alternative to the periosteal flap for reconstruction of the lateral canthal ligament when use of the lateral orbital periosteum is not feasible.

Rhodomonas salina is a phototrophic marine flagellate. We examined the ultrastructure of R. salina with particular attention to the flagellar apparatus by transmission electron microscopy and compared it with that of other cryptomonads reported previously. The major components of the flagellar apparatus in R. salina were a keeled rhizostyle (Rhs), a striated fibrous root (SR), a SR-associated microtubular root (SRm), a mitochondrion-associated lamella (ML), and three types of microtubular roots (9r, 4r, and 2r). The keeled Rhs originated near the proximal end of the dorsal basal body, passed near the nucleus and dissociated at the posterior end of the cell. The SR and SRm originated between two basal bodies and laterally extended to the right side of the cell. The ML originated between two basal bodies and extended to the left side of the cell. The 9r originated between the ventral basal body and the Rhs and extended toward the anterior dorsal lobe of the cell. The 4r originated near the 9r and extended toward the dorsal lobe with the 2r, which originated between two basal bodies. Here, the flagellar apparatus in R. salina is described, and the ultrastructure of the flagellar apparatus is compared among cryptomonad species.

Hydrogels are among the most promising biologic materials in recent technology with numerous desired applications, including serving as biosensors, drug delivery vehicles, and tissue-engineered pro...

An aqua-coordinated lamellar net [Zn(5-NH2-1,3-bdc)(H2O)] (1, 5-NH2-1,3-H2bdc = 5-amino-1,3-benzenedicarboxylic acid) has been found to undergo a reversible stimuli-responsive 2D-to-2D crystal-to-crystal transformation with a water-free bilayered-lamellar net [Zn(5-NH2-1,3-bdc)] (1′) upon removal and rebinding of aqua ligands, whereas a 2D porous pillared-bilayer [Zn2(5-NH2-1,3-bdc)2(NI-bpy-44)]·DMF (2, NI-bpy-44 = N-(pyridin-4-yl)-4-(pyridin-4-yl)-1,8-naphthalimide) has been tailored by introducing NI-bpy-44 to replace the coordinated aqua ligands. Pillared-bilayer 2 displayed a moderate CO2 uptake of 79.1 cm3 g−1 STP at P/P0 = 1 and 195 K with an isosteric heat of CO2 adsorption (Qst) of 37.0 kJ mol−1 at zero-loading. It is noteworthy that the water suspensions of 1 and 2 both displayed good fluorescence performances, which were effectively quenched by Fe3+, MnO4−, and Cr2O72− ions and shifted to long wavelengths by Fe3+, Al3+, and Cr3+, even with the coexistence of equal amounts of most other interfering ions. Taking the Stern–Volmer quenching constant, limit of detection, quenching efficiency, anti-interference ability, and visual observation into consideration, it is clear that both 1 and 2 are promising and excellent fluorescent sensors for highly sensitive detection of Fe3+, MnO4−, and Cr2O72−.

The rapid transmission of vaccinia virus (VACV) in vivo is thought to be closely related to the cell migration induced by it. Cell migration involved in dynamic changes of cell-substrate adhesion and actin cytoskeleton organization, which can influence by the micro/nano-scale topographic structures that cells are naturally exposed to via contact guidance. However, migration behaviors of VACV-infected cells exposed to topographic cues are still unknown. Herein, we designed an open chip with microgrooved poly(dimethyl siloxane) (PDMS) substrate to explore the topography roles in VACV-induced cell migration. Differed from the random cell migration observed in traditional scratch assay on planar substrate, VACV-infected cells had a tendency to persistently migrate along the axis parallel to microgroove with increased velocity. Moreover, infected cells exhibited a dominant elongated protrusion aligned to the micro-grating axis compare to the shorter lamella extended in any direction on smooth substrate. Interestingly, the Golgi complex preferred to relocate behind the nucleus confined within the micro-grating axis in majority of infected migratory cells. The directional polarization of cells embodied in protrusion formation and Golgi reorientation was responsible for the directionally persistent migration behaviors induced by VACV on microgrooved substrate. Infected cells response to substrate topography, causing the actin-filled stretched protrusion containing numerous virions and accelerated movement is likely to facilitate direct and rapid spread of VACV. This work opens a window for us to understand the migration behaviors of infected cells in vivo, and also provides a cue for revealing the relationship between virus-induced cell migration and virus rapid spread.

This study reports the complete mitochondrial genome of the Capsaloides cristatus (Monogenea: Capsalidae) collected from the gill lamella of Istiophorus platypterus. The total length of the mitogen...

PURPOSE The purpose of this study was to describe a novel device that may serve as an alternative to Descemet membrane endothelial keratoplasty (DMEK) for the treatment of chronic corneal edema. METHODS The EndoArt (EyeYon Medical, Israel) is a flexible, 50-μm thin artificial endothelial layer that matches the cornea's posterior curvature and functions as a fluid barrier at the posterior stroma, replacing the diseased endothelium. Similar to a DMEK approach, it is implanted into the anterior chamber, carefully positioned on the posterior stroma, and secured using an air-gas mixture. Two patients with chronic corneal edema resulting from endothelial decompensation underwent implantation of the new artificial lamella. RESULTS In patient 1, the central corneal thickness (CCT) decreased from 730 μm preoperatively to 593 μm at 1 day postoperatively. In patient 2, the CCT decreased from 761 μm preoperatively to 487 μm at 1 day postoperatively. Both patients reported high satisfaction and an overall brighter visual quality. Although dislocation of the lamella occurred in both cases, the CCT was promptly restored after a repositioning procedure and remained stable at the 17-month follow-up (CCT of 526 and 457 μm for patients 1 and 2, respectively). In contrast to DMEK donor tissue, the artificial lamella is sufficiently robust to allow easy intraocular manipulation without the risk of damaging the implant. It is easily removable and does not require any immunosuppressive treatment because of its nonbiological nature. CONCLUSIONS Implantation of the EndoArt led to rapid corneal deturgescence and CCT restoration, presenting a possible option for patients with chronic corneal edema.

To find out the critical damage temperature of tea leaf, a new method of subzero treatment was provided by fitting the air temperature data from six heavy frost events. Furthermore, the study explored the characteristics of Fuding Dabai tea plant response to low temperature stress of 2, 0, −2, −4, −8, −10 and −15 °C by observing the microstructure and ultrastructure changes of the leaves. All samples were collected in an ambient temperature of 8.6 °C which served as control. Scanning electron microscopy (SEM) and transmission electron microscopy (TEM) were used to observe the microstructure and ultrastructure of stomata and mesophyll. SEM observation results indicated that stomata of tea leaves have an obvious low temperature stress when the temperature was lower than −2 °C. The extent of opening of the stomata increased and enhanced guard cell protection of tea leaves against cold injury. However, dehydration, shrinkage and deformation of cells occurred as the temperature decreased from −2 °C to −15 °C. TEM observations showed that the cell nucleus, cell walls, chloroplasts and mitochondria all had normal structure at a temperature of 8.6 °C where the membrane and granum lamella were clearly visible. As the temperature decreased to −2 °C, the membrane system of tea leaf was the first to be damaged and the cell walls became fuzzy. Therefore, the leaf microstructure and ultrastructure showed obvious changes at −2 °C, which might define the critical damage temperature for freeze damage of Fuding Dabai tea. Control strategy based this critical damage temperature is useful for wind machine frost protection in tea fields within the Yangtze River region. Keywords: Frost damage, Freeze injury, Chill injury, Frost protection

Objective: Our objective is to show how the use of Computer Tomography (CT) multiplanar displays (MPR) and 3D Stereoscopic Imaging (3DSI), can provide precise anatomical landmarks to identify the location of the Anterior Ethmoid Artery (AEA) during endoscopic sinus surgery (FESS). Materials and Methods: The study comprised 48 patients, representing 96 nasal cavity/sinus sides. The anterior ethmoid foramen was the landmark that defined the entry of the AEA into the ethmoid on CT MPR and 3DSI, and anatomic relationships closely related to to the AEA were recorded. Results: The anterior ethmoidal foramen (AEF) was associated with the bulla lamella (BL) in 46.9% of cases, and the basal lamella of the middle turbinate (BLMT) in 31.6%. In 7.1% of cases, the AEF was associated with both lamellae. Similarly, at the AES, the AEF was associated with the BL and BLMT in 72.5% and 46.9% of cases, respectively. In 29.6%, the AEA was associated with both lamellae. In 48%, the AEF and AES were at or in the bone of the ethmoidal roof/ skull base. In 48%, the AEA, the AEF, and the AES were in the same coronal plane, indicating a straight horizontal course across the ethmoid. In the remaining samples, the AES was anterior to the AEF, indicating an oblique course of the AEA. Conclusion: This study demonstrates that 3DSI imaging provides improved localization of the AEA, and establishes recognizable anatomic landmarks for endoscopically guided surgery, thus, preventing inadvertent complications.

Cell migration entails a plethora of activities combining the productive exertion of protrusive and contractile forces to allow cells to push and squeeze themselves through cell clumps, interstitial tissues or tissue borders. All these activities require the generation and turnover of actin filaments that arrange into specific, subcellular structures. The most prominent structures mediating the protrusion at the leading edges of cells include lamellipodia and filopodia as well as plasma membrane blebs. Moreover, in cells migrating on planar substratum, mechanical support is being provided by an additional, more proximally located structure termed the lamella. Here, we systematically dissect the literature concerning the mechanisms driving actin filament nucleation and elongation in the best-studied protrusive structure, the lamellipodium. Recent work has shed light on open questions in lamellipodium protrusion, including the relative contributions of nucleation versus elongation to the assembly of both individual filaments and the lamellipodial network as a whole. However, much remains to be learned concerning the specificity and relevance of individual factors, their cooperation and their site-specific functions relative to the importance of global actin monomer and filament homeostasis.

We demonstrate how manipulating curvature in an elastic fluid lamella enables the reversible relative positioning of flat, rigid, plate-like micrometer-scale inclusions, with spacings from about a micrometer to tens of micrometers. In an experimental model comprising giant unilamellar vesicles containing solid domain pairs coexisting in a fluid membrane, we adjusted vesicle inflation to manipulate membrane curvature and mapped the interdomain separation. A two-dimensional model of the pair potential predicts the salient experimental observations and reveals both attractions and repulsions, producing a potential minimum entirely a result of the solid domain rigidity and bending energy in the fluid membrane. The impact of vesicle inflation on domain separation in vesicles containing two solid domains was qualitatively consistent with observations in vesicles containing many domains. The behavior differs qualitatively from the pure repulsions between fluid membrane domains or interactions between nanoscopic inclusions whose repulsive or attractive character is not switchable.

International audience; Cryo-transmission electron microscopy (cryo-TEM) provides access to high resolution information of adherent cell ultrastructures in a close to native environment but only volumes of less than 500 nm can be imaged. Cryo-focused ion beam (FIB) milling overcomes this obstacle through the generation of thin lamella of less than 200 nm. These lamellas can be imaged by cryo-electron tomography (cryo-ET) giving access to ultrastructural data within the volume of the imaged cells. Nevertheless, a lack of control on the positioning of the samples on the electron microscopy (EM) grids drastically constrains its throughput. The use of custom-designed micropatterned EM grids bypasses these issues by accurately positioning cells in areas that allow FIB milling followed by cryo-ET. Combined with fluorescent light microscopy in correlative light electron microscopy (CLEM) pipelines to pinpoint specific events and automated FIB milling, micropatterning of cells on EM grids has the potential of dramatically accelerating the workflow of cryo-ET. Here, a detailed description is provided of the key steps necessary to implement photomicropatterning of EM grids for improved CLEM pipelines.

Many cell regulatory systems implicate nonlinearity and redundancy among components. The regulatory network governing lamellipodial and lamellar actin structures is prototypical of such a system, containing tens of actin-nucleating and -modulating molecules with functional overlap and feedback loops. Due to instantaneous and long-term compensation, phenotyping the system response to perturbation provides limited information on the roles the targeted component plays in the unperturbed system. Accordingly, how individual actin regulators contribute to lamellipodial dynamics remains ambiguous. Here, we present a perturbation-free reconstruction of cause-effect relations among actin regulators by applying Granger-causal inference to constitutive image fluctuations that indicate regulator recruitment as a proxy for activity. Our analysis identifies distinct zones of actin regulator activation and of causal effects on filament assembly and delineates actin-dependent and actin-independent regulator roles in controlling edge motion. We propose that edge motion is driven by assembly of two independently operating actin filament systems. A record of this paper’s Transparent Peer Review process is included in the Supplemental Information.

Protein phosphorylation is a fundamental post-translational modification in all organisms. In photoautotrophic organisms, protein phosphorylation is essential for the fine-tuning of photosynthesis. The reversible phosphorylation of the photosystem II (PSII) core and the light-harvesting complex of PSII (LHCII) contribute to the regulation of photosynthetic activities. Besides the phosphorylation of these major proteins, recent phosphoproteomic analyses have revealed that several proteins are phosphorylated in the thylakoid membrane. In this study, we utilized the Phos-tag technology for a comprehensive assessment of protein phosphorylation in the thylakoid membrane of Arabidopsis. Phos-tag SDS-PAGE enables the mobility shift of phosphorylated proteins compared with their non-phosphorylated isoform, thus differentiating phosphorylated proteins from their non-phosphorylated isoforms. We extrapolated this technique to two-dimensional (2D) SDS-PAGE for detecting protein phosphorylation in the thylakoid membrane. Thylakoid proteins were separated in the first dimension by conventional SDS-PAGE and in the second dimension by Phos-tag SDS-PAGE. In addition to the isolation of major phosphorylated photosynthesis-related proteins, 2D Phos-tag SDS-PAGE enabled the detection of several minor phosphorylated proteins in the thylakoid membrane. The analysis of the thylakoid kinase mutants demonstrated that light-dependent protein phosphorylation was mainly restricted to the phosphorylation of the PSII core and LHCII proteins. Furthermore, we assessed the phosphorylation states of the structural domains of the thylakoid membrane, grana core, grana margin, and stroma lamella. Overall, these results demonstrated that Phos-tag SDS-PAGE is a useful biochemical tool for studying in vivo protein phosphorylation in the thylakoid membrane protein. Supplementary Information The online version of this article (10.1007/s11120-020-00803-1) contains supplementary material, which is available to authorized users.

This article examines a previously unpublished gold lamella of unknown provenance, datable on palaeographical grounds to the 1st century BCE, give-or-take a half century, either side. The tablet preserves three words written in Greek letters that may contain a GrecoPersian formula of protection in the afterlife for its bearer, Abalala, a name of pre-Islamic extraction. The study compares the formula with those on a number of shorter ‘Orphic’ gold lamellae to show that the tiny piece represents a ‘Totenpaß’ for the beneficent dead, rather than a protective charm (phylactery) with the usual voces magicae, although the distinction between magic words and meaningful text is not always clear in such instances.

Current alternatives for xenograft use in correction of lower eyelid retraction are reviewed, together with strategies for patient evaluation, typical surgical techniques, and postoperative management.

In this study, 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC), 1,2-dioleoyl-3-dimethylammonium-propane (DAP) and amphiphilic block copolymer poly(2-(dimethylamino)ethyl methacrylate)-b-poly(lauryl methacrylate) (PLMA-b-PDMAEMA) in the weight ratios: 1:0:0.03, 1:0.1:0.03, 1:0.3:0.03, 1:0.5:0.03, 1:0.7:0.03, and1:1:0.03, were used for the design and development of hybrid polymer-lipid nanostructures. The polymer-lipid nanostructures were prepared using the aqueous heat method. The pure lipid systems were used as reference systems. Differential Scanning Calorimetry (DSC) was utilized to investigate the interactions between the components. The presence of the polymeric guest caused different orientation of the polar groups of phospholipids, as the DSC thermotropic values revealed. The changes of the physicochemical characteristics of the systems with the increase of the weight ratio of DAP are more intense in comparison to those without this lipid due to the presence of the block copolymer, which contributes significantly to the repulsive interactions between the vesicles as a result of its positive charge. The only exception is the DSPC:DAP:PLMA-b-PDMAEMA nanosystem at 1:0.5:0.03 weight ratio, where the size of the polymer/lipid structures (Dh≈320 nm) increased and the ζ-potential decreased in absolute values in comparison with the systems with lower weight ratio of the DAP lipid. Cryogenic Transmission Electron Microscopy (cryo-TEM) images showed different structures which strongly depend on the composition of the system, ranging from lamella structures, disc forms, and “spaghetti” morphologies. Based on toxicity studies, it was found all samples are biocompatible and show a dose-dependent cytotoxicity on HEK293 (Human Embryonic Kidney 293 cell line). In conclusion, we developed polymer/lipid nanostructures with knowledge of their structural and physicochemical characteristics, useful as drug or antigen delivery platforms.

This article briefly reviews recent advances in nano-scale and micro-scale assessments of primary cell wall structure, mechanical behaviors and expansive growth. Cellulose microfibrils have hydrophobic and hydrophilic faces which may selectively bind different matrix polysaccharides and adjacent microfibrils. These distinctive binding interactions may guide partially aligned cellulose microfibrils in primary cell walls to form a planar, load-bearing network within each lamella of polylamellate walls. Consideration of expansive growth of cross-lamellate walls leads to a surprising inference: side-by-side sliding of microfibrils may be a key rate-limiting physical step, potentially targeted by specific wall loosening agents. Atomic force microscopy shows different patterns of microfibril movement during force-driven extension versus enzymatic loosening. Consequently, simulations of cell growth as elastic deformation of isotropic cell walls may need to be augmented to incorporate the distinctive behavior of growing cell walls.

With their ability to survive complete desiccation, resurrection plants are a suitable model system for studying the mechanisms of drought tolerance. In the present study, we investigated desiccation-induced alterations in surface topography of thylakoids isolated from well-hydrated, moderately dehydrated, severely desiccated and rehydrated Haberlea rhodopensis plants by means of atomic force microscopy (AFM), electrokinetic and optical measurements. According to our knowledge, so far, there were no reports on the characterization of surface topography and polydispersity of thylakoid membranes from resurrection plants using AFM and dynamic light scattering. To study the physicochemical properties of thylakoids from well-hydrated H. rhodopensis plants, we used spinach thylakoids for comparison as a classical model from higher plants. The thylakoids from well-hydrated H. rhodopensis had a grainy surface, significantly different from the well-structured spinach thylakoids with distinct grana and lamella, they had twice smaller cross-sectional area and were 1.5 times less voluminous than that of spinach. Significant differences in their physicochemical properties were observed. The dehydration and subsequent rehydration of plants affected the size, shape, morphology, roughness and therefore the structure of the studied thylakoids. Drought resulted in significant enhancement of negative charges on the outer surface of thylakoid membranes which correlated with the increased roughness of thylakoid surface. This enhancement in surface charge density could be due to the partial unstacking of thylakoids exposing more negatively charged groups from protein complexes on the membrane surface that prevent from possible aggregation upon drought stress.

Annulus defect is associated with reherniation and disc degeneration after discectomy; currently there is no effective treatment that addresses this problem. The annulus is a hierarchical lamellar structure, where each lamella consists of aligned collagen fibres, which are parallel and tilted at 30° to the spinal axis. In this study, a biomimetic biodegradable scaffold consisting of multilamellar nano/microfibres, sharing nanotopography and microporosity similar to the native lamellar structure, was assessed in a porcine model, aided by sealing with fascia and medical glue and subsequent suture fixation. After 6- and 12-week observation, we found that this treatment restored nucleus volume and slowed down disc degeneration, as indicated by magnetic resonance imaging of T1/T2-weighted, T2-mapping, T1-ρ imaging. Histological analysis showed aligned collagen fibres organized in the scaffold and integrated with surrounding native annulus tissue. The autologous bone marrow concentrate-seeded scaffolds showed slightly earlier collagen fibre formation at 6 weeks. This novel treatment could efficiently close the annulus defect with newly formed, organized and integrated collagen fibres in a porcine model. Copyright © 2016 John Wiley & Sons, Ltd.

A new design for syrup and juice clarifiers is presented. The design takes advantage of the considerably improved performance of clarifiers incorporating lamella plates, and the reasons for the improvement are outlined. Computational fluid dynamics (CFD) work done to simulate the performance is summarised. This design enables the residence time to be dramatically reduced and the simplified design leads to cheaper and better clarifiers. Practical experience with factory scale units is described, confirming the good flow characteristics. The results of preliminary test work on a factory syrup clarifier are presented, which is also shown to operate efficiently as a phosphatation clarifier. In addition the performance of a full-scale juice clarifier has been evaluated and compared with the performance of a Rapidorr clarifier. This work confirms the considerable advantages which this type of design provides, in realising substantial reductions in residence time, capital costs and operating costs.

Cytotoxicity of nickel oxide nanoparticles(NiO NPs) with average diameter of 20 nm were investigated on cultured Chlorella vulgaris. Alga growth-inhibition tests were taken and ultrastructure changes of the microalgae were characterized with transmission electron microscopy(TEM). The biological interface conversion effect between NiO nanoparticles and Chlorella vulgaris were studied by X-ray diffraction(XRD), high-resolution transmission electron microscopy(HRTEM) and X-ray photoelectron spectroscopy(XPS). The results indicated that the NiO nanoparticles had severe inhibitory effect on the growth of microalgae, with a 96 h EC50 value of 31.4 mg/L. Under the exposure to NiO NPs suspensions, Chlorella vulgaris cells showed plasmolysis with a shriveled cell shape, disrupted plasma mem-brane, leaked cytosol and disordered thylakoid grana lamella. The NiO NPs were aggregated and partially reduced to Ni0 inside the Chlorella vulgaris. The bioaccumulation and bio-reduction ability of Chlorella vulgaris provide us with a possible strategy of remediation of aquatic pollution conducted by toxic metal oxide nanoparticles.

ABSTACTCryo-electron tomography (cryo-ET) provides a promising technique to study high resolution structures of macromolecules in situ, opening a new era of structural biology. One major bottleneck of this technique is to prepare suitable cryo-lamellas of cell/tissue samples. The emergence of cryo-focused ion beam (cryo-FIB) milling technique provides a good solution of this bottleneck. However, there are still large limitations of using cryo-FIB to prepare cryo-lamella of tissue specimen because the thickness of tissue increases the difficulty of specimen freezing and cryo-FIB milling. Here we report a new workflow, VHUT-cryo-FIB (Vibratome - High pressure freezing - Ultramicrotome Trimming – cryo-FIB), aiming for efficient preparation of frozen hydrated tissue lamella for subsequent cryo-ET data collection. This workflow includes tissue slicing using vibratome, high pressure freezing, ultramicrotome cryo-trimming, cryo-FIB milling and the subsequent cryo-electron microscopy (cryo-EM). The modification of equipment in this workflow is highly eliminated. We developed two strategies with a special cryo-holder tip or carrier for loading cryo-lamella into side entry cryo-holder or Autoloader catridge. We tested this workflow using the tissue sample of rat skeleton muscle and spinach leaf and collected high quality cryo-ET tilt series, which enabled us to obtain an in situ structure of spinach ribosome by sub-tomogram averaging.

Cryo-Electron Tomography (cryo-ET) is emerging as a revolutionary method for resolving the structure of macromolecular complexes in situ. However, sample preparation for in situ Cryo-ET is labour-intensive and can require both cryo-lamella preparation through cryo-Focused Ion Beam (FIB) milling and correlative light microscopy to ensure that the event of interest is present in the lamella. Here, we present an integrated cryo-FIB and light microscope setup called the Photon Ion Electron microscope (PIE-scope) that enables direct and rapid isolation of cellular regions containing protein complexes of interest. Specifically, we demonstrate the versatility of PIE-scope by preparing targeted cryo-lamellae from subcellular compartments of neurons from transgenic Caenorhabditis elegans and Drosophila melanogaster expressing fluorescent proteins. We designed PIE-scope to enable retrofitting of existing microscopes, which will increase the throughput and accuracy on projects requiring correlative microscopy to target protein complexes. This new approach will make cryo-correlative workflow safer and more accessible.

Motile cilia are oscillating nanomachinery that are essential for cell locomotion and the generation of extracellular fluid flow. They typically comprise an axoneme with a 9+2 microtubule structure with dynein arms on each microtubule doublet that generate force by sliding between adjacent microtubules. These structures are efficient for motility and have therefore been conserved in cellular-level aquatic microflow throughout eukaryotic evolution. Diversity in the ciliary structures is seen in the basic microtubule arrangements but also in the bundling of multiple cilia, such as compound cilia. Studies on the structural diversity of cilia shed light on the mechanisms underlying ciliary formation and the biological adaptation to different aquatic environments. Ctenophores (comb jellies) have eight rows of iridescent comb plates that facilitate locomotion. A comb plate consists of tens of thousands of cilia to form a “puddle-like” ciliary plate with lengths of up to 1 mm. It exhibits structural color by light diffraction and is reported to have properties similar to photonic crystals. Here we identified a protein specific to the construction of comb plates (CTENO64). This protein is specifically present in ctenophore species and is localized on the compartmenting lamella (CL), which is the structure connecting adjacent cilia on the comb plates. Knockdown of the CTENO64 gene caused loss or misformation of CLs, resulting in aberrant waveforms in the comb plates. Our findings provide the initial characterization of a protein involved in the bundling of cilia into giant compound ciliary comb plates.

The most important structure of the cornea is stroma, which has layer tissue and consists of 200 stromal lamella constructed by parallel collagen fibrils (Fig. 38.1).

Annulus fibrosus (AF) impairment is associated with reherniation, discogenic pain, and disc degeneration after surgical partial discectomy. Due to a limited intrinsic healing capacity, defects in the AF persist over time and it is hence necessary to adopt an appropriate strategy to close and repair the damaged AF. In this study, a cell-free biodegradable scaffold made of polycaprolactone (PCL), electrospun, aligned microfibers exhibited high levels of cell colonization, alignment, and AF-like extracellular matrix deposition when evaluated in an explant culture model. The biomimetic multilayer fibrous scaffold was then assessed in an ovine model of AF impairment. After 4 weeks, no dislocation of the implants was detected, and only one sample out of six showed a partial delamination. Histological and immunohistochemical analyses revealed integration of the implant with the surrounding tissue as well as homogeneously aligned collagen fiber organization within each lamella compared to the disorganized and scarcer fibrous tissue in a randomly organized control fibrous scaffold. In conclusion, this biomimetic electrospun implant exhibited promising properties in terms of AF defect closure, with AF-like neotissue formation that fully integrated with the surrounding ovine tissue. Accepted version

In this paper, the three nicotinic acid-based amphiphiles, sodium 6-dodecylnicotinic glycinate, sodium 6-dodecylnicotinic alaninate, and sodium 6-dodecylnicotinic valinate, were synthesized, and the effect of the headgroup on the self-assembly properties was investigated. The interfacial properties were determined by tensiometry measurement. A steady-state fluorescence technique was used to investigate the microenvironment of the self-assemblies. The amphiphiles form vesicles in water, and the size of the aggregates decreases with increase in the hydrophobicity of the headgroup. Gel permeation chromatography and kinetic studies were performed to establish the entrapping and releasing properties of the vesicles. The arrangement of the hydrocarbon chains in the lamella was examined by X-ray diffraction study. Circular dichroism spectra suggested formation of chiral structure through aggregation of the chiral amphiphiles. It has been demonstrated that the amphiphiles have the feasibility to be developed for ...

Background and Aims In the present study, we show that development of endodermis and exodermis is sensitively regulated by water accessibility. As cadmium (Cd) is known to induce xeromorphic effects in plants, maize roots were exposed also to Cd to understand the developmental process of suberin lamella deposition in response to a local Cd source. Methods In a first experiment, maize roots were cultivated in vitro and unilaterally exposed to water-containing medium from one side and to air from the other. In a second experiment, the roots were placed between two agar medium layers with a strip of Cd-containing medium attached locally and unilaterally to the root surface. Key Results The development of suberin lamella (the second stage of exodermal and endodermal development) started asymmetrically, preferentially closer to the root tip on the side exposed to the air. In the root contact with Cd in a spatially limited area exposed to one side of the root, suberin lamella was preferentially developed in the contact region and additionally along the whole length of the root basipetally from the contact area. However, the development was unilateral and asymmetrical, facing the treated side. The same pattern occurred irrespective of the distance of Cd application from the root apex. Conclusions These developmental characteristics indicate a sensitive response of root endodermis and exodermis in the protection of vascular tissues against abiotic stresses.

We present covalently self-assembled peptide hollow nanocapsule and peptide lamella. These biomimetic dityrosine peptide nanostructures are synthesized by one-step photopolymerization of a tyrosine-rich short peptide without the aid of a template. This simple approach offers direct synthesis of fluorescent peptide nanocages and free-standing thin films. The simple crosslinked peptide lamella films provide robust mechanical properties with an elastic modulus of approximately 30 GPa and a hardness of 740 MPa. These nanostructures also allow for the design of peptidosomes. The approach taken here represents a rare example of covalent self-assembly of short peptides into nano-objects, which may be useful as microcompartments and separation membranes.

The development of cryo-focused ion beam (cryo-FIB) for the thinning of frozen-hydrated biological specimens enabled cryo-electron tomography (cryo-ET) analysis in unperturbed cells and tissues. However, the volume represented within a typical FIB lamella constitutes a small fraction of the biological specimen. Retaining low-abundance and dynamic subcellular structures or macromolecular assemblies within such limited volumes requires precise targeting of the FIB milling process. In this study, we present the development of a cryo-stage allowing for spinning-disk confocal light microscopy at cryogenic temperatures and describe the incorporation of the new hardware into existing workflows for cellular sample preparation by cryo-FIB. Introduction of fiducial markers and subsequent computation of three-dimensional coordinate transformations provide correlation between light microscopy and scanning electron microscopy/FIB. The correlative approach is employed to guide the FIB milling process of vitrified cellular samples and to capture specific structures, namely fluorescently labeled lipid droplets, in lamellas that are 300 nm thick. The correlation procedure is then applied to localize the fluorescently labeled structures in the transmission electron microscopy image of the lamella. This approach can be employed to navigate the acquisition of cryo-ET data within FIB-lamellas at specific locations, unambiguously identified by fluorescence microscopy.

Bone tissue engineering necessitates a three-dimensional porous scaffold design to closely mimic the in vivo physiological biomimetic microenvironment with good mechanical properties to support the adhesion and proliferation of osteoblasts. In this study, a silk fibroin/cellulose nanowhiskers–chitosan (SF/CNW–CS) composite scaffold with excellent mechanical properties and biological compatibility was prepared by layer-by-layer assembly of CNW and CS onto a porous SF scaffold fabricated by freeze-drying. The morphology, assembly construct, and mechanical properties of the SF/CNW–CS composite scaffold were characterized using different technical methods. The hierarchical lamellar structure was formed by assembly of CNW and CS onto the SF lamella. Increasing the number of CNW and CS assembly layers to 108 slightly decreased the porosity from 86 to 78 % and simultaneously changed the compressive stress–strain curves from soft to hard tissue-like properties with an increase in compressive modulus from 2.5 to 12.5 MPa. Human MG-63 osteosarcoma cells were further cultured on the SF/CNW–CS scaffold to evaluate their suitability for bone tissue engineering. The results indicated that the SF/CNW–CS composite scaffold supported cell proliferation and promoted the levels of biomineralization-relevant alkaline phosphatase activity and osteocalcin expression over those for a porous SF scaffold. Taken together, our results indicate that the porous SF/CNW–CS composite material will be a promising tissue engineering scaffold for bone generation and implantation.

The Berezan lead letter is a document of paramount importance with regard to several epigraphic questions (first of all, the writing on lead platters) and to many historical issues too (the Greek presence, mainly Milesian, within ancient Scythia and the diffusion of writing in this area; the ionic dialect; the relationship between literacy and commerce; archaic Greek prose; Greek epistolography; the right of seizure and the regulation of syle); moreover, it is a human life story that vividly shows us the dramatic situation of Achillodoros who, harassed by Matasys, sends a heartfelt letter to two recipients, his son Protagoras and Anaxagoras.

The neural fat-body sheath surrounding the abdominal ventral nerve cord of Carausius morosus has been examined by light and electron microscopy. A perineural chamber between the ventral nerve cord and the sheath is present in the ganglionic regions, but in the interconnective regions the sheath directly covers the neural lamella. The sheath is differentiated into secretory cells with abundant granular endoplasmic reticulum and many mitochondria and storage cells with lipid droplets and granules presumed to be glycogen. The whole sheath is pervaded with tracheoblast cells and associated tracheal and tracheolar tubules. Recent evidence suggests that this sheath may have little power of ionic regulation. The function of the sheath, deduced from the fine structure described here, appears to be to serve the nutritional needs of the central nervous system. Why a sheath of this type appears to be confined to herbivorous insects with unusual haemolymph cationic balance is still unexplained.

The epidermis of Eisenia is covered by a cuticle and rests on a basement lamella. The cuticle, which is resistant to a variety of enzymes, is composed of non-striated, bundles of probable collagen fibers that are orthogonally oriented and are embedded in a proteoglycan matrix. The basement lamella consists of striated collagen fibers with a 560 A major periodicity. Proximity and morphology suggest that the epidermis may contribute to both the cuticle and the basement lamella — that is, the single tissue may synthesize at least two types of collagen. The epidermis is a pseudostratified epithelium containing three major cell types (columnar, basal and gland) and a rare fourth type with apical cilia. The esophagus is lined by a simple cuticulated epithelium composed predominantly of a single cell type, which resembles the epidermal columnar cell. Rare gland cells occur in the esophageal epithelium, but basal cells are lacking.

Abl family nonreceptor tyrosine kinases regulate changes in cell shape and migration. Abl2 localizes to dynamic actin-rich protrusions, such as lamellipodia in fibroblasts and dendritic spines in neurons. Abl2 interactions with cortactin, an actin filament stabilizer, are crucial for the formation and stability of actin-rich structures, but Abl2:cortactin-positive structures have not been characterized with high spatiotemporal resolution in cells. Using total internal reflection fluorescence microscopy, we demonstrate that Abl2 colocalizes with cortactin at wave-like structures within lamellum and lamellipodium tips. Abl2 and cortactin within waves are focal and transient, extend to the outer edge of lamella, and serve as the base for lamellipodia protrusions. Abl2-positive foci colocalize with integrin β3 and paxillin, adhesive markers of the lamellum–lamellipodium interface. Cortactin-positive waves still form in Abl2 knockout cells, but the lamellipodium size is significantly reduced. This deficiency is restored following Abl2 reexpression. Complementation analyses revealed that the Abl2 C-terminal half, which contains domains that bind actin and microtubules, is necessary and sufficient for recruitment to the wave-like structures and to support normal lamellipodium size, while the kinase domain–containing N-terminal half does not impact lamellipodium size. Together, this work demonstrates that Abl2 is recruited with cortactin to actin waves through cytoskeletal interactions to promote lamellipodium extension.

Plant virus was a kind of organism lived depending on infecting viable host cell and propagated their posterity by replicating its hereditary nucleotide, transcripting into protein, assembling protein and nucleotide into virion (Ortin and Parra, 2006; Sanfacon, 2005). Viral infection usually induces remodeling of host cell, especially endoplasmic reticulum (ER) for generating membrane packed viral factory. During the infection of Bymovirus, a kind of membranous body (MB) was generated in host cells, which is thought as an ER aggregate. In present study we performed a study on Wheat yellow mosaic virus (WYMV) induced MB by several transmission electron microscopy (TEM) based methods, including cytological observation, component analysis by immuno-gold labeling and structural analysis by electron tomography (ET). WYMV infection induced at least two morphologies of MB, including the lamella dominated morphology (lamella-MB) looked like sprawling cirrus, and the tubule dominated morphology (tubule-MB) looked like latticed network. MB was verified composing of ER as revealed by immuno-gold labeling by antibody against endoplasmic reticulum (ER) retention signal as well as by detailed observation of MB construction modules as double layer membrane. By immuno-gold labeling, both two MB morphologies (lamella-MB and tubule-MB) had same components in viral derived protein and membrane origination (from ER). Structural analysis by ET reconstruction revealed the organization of ER in MB. Lamella-MB was composed of cesER like structures arranged irregularly whereas tubule-MB was composed of tubER like structures arranged regularly. This study provided insights into the structural details in how Bymovirus utilizing host membrane system.