Organized Cellular Structure Research Articles

Information–Thermodynamic Method for the Study of Proliferation of Organized Cellular Structure

The aim of the article was to develop an innovative method for the study of cell proliferation based on the information-thermodynamic approach, including the mathematical ratio-the entropy of cell proliferation and an algorithm for the calculation of fractal dimension of the cellular structure. Approbation of this method with pulsed electromagnetic impact on culture in vitro was implemented. It is shown on the basis of experimental data that the organized cellular structure of juvenile human fibroblasts is a fractal. The method makes it possible to determine the stability of the effect on cell proliferation. The prospects for the application of the developed method are discussed.

Transcriptomic Characterization of Bradyrhizobium diazoefficiens Bacteroids Reveals a Post-Symbiotic, Hemibiotrophic-Like Lifestyle of the Bacteria within Senescing Soybean Nodules.

The transcriptional activity of Bradyrhizobium diazoefficens isolated from soybean nodules was monitored over the period from symbiosis to late plant nodule senescence. The bacteria retained a near constant level of RNA throughout this period, and the variation in genes demonstrating increased, decreased, and/or patterned transcriptional activity indicates that the bacteria are responding to the changing environment within the nodule as the plant cells progress from an organized cellular structure to an unorganized state of internal decay. The transcriptional variation and persistence of the bacteria suggest that the bacteria are adapting to their environment and acting similar to hemibiotrophs, which survive both as saprophytes on live plant tissues and then as necrophytes on decaying plant tissues. The host plant restrictions of symbiosis make B. diazoefficiens a highly specialized, restricted hemibiotroph.

3D Bioprinted Human Skeletal Muscle Constructs for Muscle Function Restoration

A bioengineered skeletal muscle tissue as an alternative for autologous tissue flaps, which mimics the structural and functional characteristics of the native tissue, is needed for reconstructive surgery. Rapid progress in the cell-based tissue engineering principle has enabled in vitro creation of cellularized muscle-like constructs; however, the current fabrication methods are still limited to build a three-dimensional (3D) muscle construct with a highly viable, organized cellular structure with the potential for a future human trial. Here, we applied 3D bioprinting strategy to fabricate an implantable, bioengineered skeletal muscle tissue composed of human primary muscle progenitor cells (hMPCs). The bioprinted skeletal muscle tissue showed a highly organized multi-layered muscle bundle made by viable, densely packed, and aligned myofiber-like structures. Our in vivo study presented that the bioprinted muscle constructs reached 82% of functional recovery in a rodent model of tibialis anterior (TA) muscle defect at 8 weeks of post-implantation. In addition, histological and immunohistological examinations indicated that the bioprinted muscle constructs were well integrated with host vascular and neural networks. We demonstrated the potential of the use of the 3D bioprinted skeletal muscle with a spatially organized structure that can reconstruct the extensive muscle defects.

Neonatal Murine Cochlear Explant Technique as an In Vitro Screening Tool in Hearing Research.

While there have been remarkable advances in hearing research over the past few decades, there is still no cure for Sensorineural Hearing Loss (SNHL), a condition that typically involves damage to or loss of the delicate mechanosensory structures of the inner ear. Sophisticated in vitro and ex vivo assays have emerged in recent years, enabling the screening of an increasing number of potentially therapeutic compounds while minimizing resources and accelerating efforts to develop cures for SNHL. Though homogenous cultures of certain cell types continue to play an important role in current research, many scientists now rely on more complex organotypic cultures of murine inner ears, also known as cochlear explants. The preservation of organized cellular structures within the inner ear facilitates the in situ evaluation of various components of the cochlear infrastructure, including inner and outer hair cells, spiral ganglion neurons, neurites, and supporting cells. Here we present the preparation, culture, treatment, and immunostaining of neonatal murine cochlear explants. The careful preparation of these explants facilitates the identification of mechanisms that contribute to SNHL and constitutes a valuable tool for the hearing research community.

Neonatal Murine Cochlear Explant Technique as an <em>In Vitro</em> Screening Tool in Hearing Research

While there have been remarkable advances in hearing research over the past few decades, there is still no cure for Sensorineural Hearing Loss (SNHL), a condition that typically involves damage to or loss of the delicate mechanosensory structures of the inner ear. Sophisticated in vitro and ex vivo assays have emerged in recent years, enabling the screening of an increasing number of potentially therapeutic compounds while minimizing resources and accelerating efforts to develop cures for SNHL. Though homogenous cultures of certain cell types continue to play an important role in current research, many scientists now rely on more complex organotypic cultures of murine inner ears, also known as cochlear explants. The preservation of organized cellular structures within the inner ear facilitates the in situ evaluation of various components of the cochlear infrastructure, including inner and outer hair cells, spiral ganglion neurons, neurites, and supporting cells. Here we present the preparation, culture, treatment, and immunostaining of neonatal murine cochlear explants. The careful preparation of these explants facilitates the identification of mechanisms that contribute to SNHL and constitutes a valuable tool for the hearing research community.

CD8 T cells and Mycobacterium tuberculosis infection.

Tuberculosis is primarily a respiratory disease that is caused by Mycobacterium tuberculosis. M. tuberculosis can persist and replicate in macrophages in vivo, usually in organized cellular structures called granulomas. There is substantial evidence for the importance of CD4 T cells in control of tuberculosis, but the evidence for a requirement for CD8 T cells in this infection has not been proven in humans. However, animal model data support a non-redundant role for CD8 T cells in control of M. tuberculosis infection. In humans, infection with this pathogen leads to generation of specific CD8 T cell responses. These responses include classical (MHC Class I restricted) and non-classical CD8 T cells. Here, we discuss the potential roles of CD8 T cells in defense against tuberculosis, and our current understanding of the wide range of CD8 T cell types seen in M. tuberculosis infection.

Anisotropic poly (glycerol sebacate)-poly (ϵ-caprolactone) electrospun fibers promote endothelial cell guidance

Topographical cell guidance is utilized to engineer highly organized and aligned cellular constructs for numerous tissue engineering applications. Recently, electrospun scaffolds fabricated using poly(glycerol sebacate) (PGS) and poly(ϵ-caprolactone) (PCL) have shown a great promise to support valvular interstitial cell functions for the development of tissue engineered heart valves. However, one of the major drawbacks of PGS-PCL scaffolds is the lack of control over cellular alignment. In this work, we investigate the role of scaffold architecture on the endothelial cell alignment, proliferation and formation of organized cellular structures. In particular, PGS-PCL scaffolds with randomly oriented and highly aligned fibers with tunable mechanical properties were fabricated using electrospinning technique. After one week of culture, endothelial cells on the aligned scaffolds exhibited higher proliferation compared to those cultures on randomly oriented fibrous scaffolds. Furthermore, the endothelial cells reorganized in response to the topographical features of aligned scaffolds forming highly organized cellular constructs. Thus, topographical contact guidance, provided by aligned PGS-PCL scaffolds, is envisioned to be useful in developing cellular structures for vascular tissue engineering.

Magnetic susceptibility induced white matter MR signal frequency shifts—experimental comparison between Lorentzian sphere and generalized Lorentzian approaches

The nature of the remarkable phase contrast in high-field gradient echo MRI studies of human brain is a subject of intense debates. The generalized Lorentzian approach (He and Yablonskiy, Proc Natl Acad Sci USA 2009;106:13558-13563) provides an explanation for the anisotropy of phase contrast, the near absence of phase contrast between white matter and cerebrospinal fluid, and changes of phase contrast in multiple sclerosis. In this study, we experimentally validate the generalized Lorentzian approach. The Generalized Lorentzian Approach suggests that the local contribution to frequency shifts in white matter does not depend on the average tissue magnetic susceptibility (as suggested by Lorentzian sphere approximation), but on the distribution and symmetry of magnetic susceptibility inclusions at the cellular level. We use ex vivo rat optic nerve as a model system of highly organized cellular structure containing longitudinally arranged myelin and neurofilaments. The nerve's cylindrical shape allowed accurate measurement of its magnetic susceptibility and local frequency shifts. We found that the volume magnetic susceptibility difference between nerve and water is -0.116 ppm, and the magnetic susceptibilities of longitudinal components are -0.043 ppm in fresh nerve, and -0.020 ppm in fixed nerve. The frequency shift observed in the optic nerve as a representative of white matter is consistent with generalized Lorentzian approach but inconsistent with Lorentzian sphere approximation.

Membrane Association via an Amino-terminal Amphipathic Helix Is Required for the Cellular Organization and Function of RNase II

The subcellular localization of the exoribonuclease RNase II is not known despite the advanced biochemical characterization of the enzyme. Here we report that RNase II is organized into cellular structures that appear to coil around the Escherichia coli cell periphery and that RNase II is associated with the cytoplasmic membrane by its amino-terminal amphipathic helix. The helix also acts as an autonomous transplantable membrane binding domain capable of directing normally cytoplasmic proteins to the membrane. Assembly of the organized cellular structures of RNase II required the RNase II amphipathic membrane binding domain. Co-immunoprecipitation of the protein from cell extracts indicated that RNase II interacts with itself. The RNase II self-interaction and the ability of the protein to assemble into organized cellular structures required the membrane binding domain. The ability of RNase II to maintain cell viability in the absence of the exoribonuclease polynucleotide phosphorylase was markedly diminished when the RNase II cellular structures were lost due to changes in the amphipathicity of the amino-terminal helix, suggesting that membrane association and assembly of RNase II into organized cellular structures play an important role in the normal function of the protein within the bacterial cell.

Planar cell polarity pathway regulates actin rearrangement, cell shape, motility, and nephrin distribution in podocytes

Glomerular podocytes are highly polarized cells characterized by dynamic actin-based foot processes (FPs). Neighboring FPs form specialized junctions, slit diaphragms (SDs), which prevent passage of proteins into the ultrafiltrate. The SD protein complex is linked to cytoskeletal actin filaments and mutations in SD proteins lead to a dramatic change in cell morphology; proteinuria is accompanied by FP retraction and loss of SD structure. Thus, organization of the podocyte cytoskeleton is tightly linked to filtration barrier function. In a variety of cell systems, cytoskeleton arrangement is regulated by the planar cell polarity (PCP) pathway. PCP signals lead to the appearance of highly organized cellular structures that support directional cell movement and oriented cell division. Derangement of the PCP pathway causes neural tube defects and cystic kidney disease in mice. Here, we establish that the PCP pathway regulates the cytoskeleton of podocytes. We identify expression of core PCP proteins in mouse kidney sections and of PCP transcripts in murine and human cultured podocytes. The pathway is functional since Wnt5a causes redistribution of PCP proteins Dishevelled and Daam1. We also show that Wnt5a treatment changes podocyte morphology, alters nephrin distribution, increases the number of stress fibers, and increases cell motility. In reciprocal experiments, siRNA depletion of the core PCP gene Vangl2 reduced the number of cell projections and decreased stress fibers and cell motility. Finally, we demonstrate direct interactions between Vangl2 and the SD protein, MAGI-2. This suggests that the PCP pathway may be directly linked to organization of the SD as well as to regulation of podocyte cytoskeleton. Our observations indicate that PCP signaling may play an important role both in podocyte development and FP cytoskeleton dynamics.

Reaction zones in highly unstable detonations

Experimental images of detonation fronts are made for several fuel-oxidizer mixtures, including hydrocarbon–air systems. Schlieren and planar laser induced fluorescence techniques are used to image both the shock configurations and the OH reaction front structure in a single experiment. The experiments are carried out in a narrow rectangular channel. The degree of instability of detonation fronts in different mixtures is evaluated by comparing calculated mixture parameters with the longitudinal neutral stability curve. The images reveal that the structure of the front increases dramatically in complexity as the mixture parameters move away from the neutral stability curve into the unstable region. Of the mixtures studied, nitrogen-diluted hydrocarbon mixtures are predicted to be the most unstable, and these show the greatest degree of wrinkling in the shock and OH fronts, with distortion occurring over a wide range of spatial scales. In the most unstable cases, separation of the shock and OH front occurs, and localized explosions in these regions are observed in a high-speed schlieren movie. This is in dramatic contrast to the weakly unstable waves that have smooth reaction fronts and quasi-steady reaction zones with no evidence of localized explosions. A key feature of highly unstable waves is very fine scale wrinkling of the OH and shock fronts, which is absent in the low-activation energy cases. This may be due to the superposition of cellular structures with a wide range of cell sizes. In contrast to soot foils, images of the OH front have a more stochastic appearance, and organized cellular structure is not as apparent.

On the reflectance of cellular cloud layers

AbstractA Monte Carlo model is used to investigate the effects on the reflectance of low cloud layers of organized cellular structure, which is often observed in such layers. It is assumed that the cloud is composed of a series of cells each of which comprises a core region surrounded by a wall region of reduced water content. It is shown that the sensitivity of reflectance to the structure is largest for wavelengths that are not absorbed by liquid water. The reflectance of the cloud layer is reduced when the water content in the regions of low water content is less than about 30% of the water content in the remainder of the layer. The effect on the cloud‐layer reflectance of organized structure in the water content is large compared with the effect of changes in the effective radius at cloud top. It is further shown that, for narrow (∼100 m) regions of low water content, estimations of the layer average reflectance based on the area weighted average of the reflectance of layer clouds with high and low water contents are significantly in error. For small solar zenith angles the regions of low water content tend to permit incident radiation to pass through the cloud, reducing the reflectance, while for large solar zenith angles the reflectance is increased by the effects of cloud shadowing.

Applications of ESE spectroscopy in the study of electron spin polarization in bacterial photosynthesis

Abstract

Electron spin polarization of photosynthetic reactants.

Photosynthesis is the conversion of the quantum energy of light into the chemical energy of complex organic molecules and organized cellular structures in plants and in some bacteria. The processes of photosynthesis span the time domain of subpicoseconds to the millennia of slow-growing trees, its study brings together such diverse disciplines as photophysics, biochemistry, botany and ecology. In the last few decades tremendous progress has been made in understanding the multivarious chemical reactions that ultimately lead to the fixation of carbon dioxide into organic substance, yet the basic mechanism underlying the conversion of photon energy into chemical energy still remains very much an enigma. These so-called primary reactions which transduce the excitation energy of excited chlorophyll pigments into the potential energy of stabilized, separated charges on electron donor and electron acceptor molecules have been studied with a variety of physical techniques, among which fast optical spectroscopy and electron paramagnetic resonance (EPR) are prominent. This review will highlight one intriguing aspect of EPR, namely that of electron spin polarization (ESP).† It will be shown that ESP of photosynthetic primary reactants offers a unique tool to gain insight in the electrostatic and magnetic interactions that make photosynthesis work. Moreover, it will become apparent that ESP in photosynthesis has several unique traits not (yet) found in ESP of photochemical reactionsin vitro. As such, it may serve as a paradigma of ESP phenomena and will present an absorbing spectacle also for EPR spectroscopists outside photosynthesis.

The action of sensitized lymphocytes on the corneal endothelium of rabbits

A host versus graft reaction is induced by the injection of sensitized lymphocytes into the anterior chamber of rabbit eyes. The reaction of these lymphocytes against the corneal endothelium is studied with S.E.M. and T.E.M. On the first and second days the transformation of lymphocytes into active cells with uropods is observed. These immunologic cells destroy the endothelium, but the endothelial cells regenerate quickly. On the sixth day there appear macrophages coming from the host, indicating that the lesions are being cleaned. These experiments present a good model for the study of the interaction between immunologic cells and an organized cellular structure.