Changes In Biophysical Properties Research Articles

Neuronal properties of pyramidal cells in lateral prefrontal cortex of the aging rhesus monkey brain are associated with performance deficits on spatial working memory but not executive function.

Age-related declines in cognitive abilities occur as early as middle-age in humans and rhesus monkeys. Specifically, performance by aged individuals on tasks of executive function (EF) and working memory (WM) is characterized by greater frequency of errors, shorter memory spans, increased frequency of perseverative responses, impaired use of feedback and reduced speed of processing. However, how aging precisely differentially impacts specific aspects of these cognitive functions and the distinct brain areas mediating cognition are not well understood. The prefrontal cortex (PFC) is known to mediate EF and WM and is an area that shows a vulnerability to age-related alterations in neuronal morphology. In the current study, we show that performance on EF and WM tasks exhibited significant changes with age, and these impairments correlate with changes in biophysical properties of layer 3 (L3) pyramidal neurons in lateral LPFC (LPFC). Specifically, there was a significant age-related increase in excitability of L3 LPFC pyramidal neurons, consistent with previous studies. Further, this age-related hyperexcitability of LPFC neurons was significantly correlated with age-related decline on a task of WM, but not an EF task. The current study characterizes age-related performance on tasks of WM and EF and provides insight into the neural substrates that may underlie changes in both WM and EF with age.

A change in biophysical properties accompanies heterochromatin formation in mouse embryos.

The majority of our genome is composed of repeated DNA sequences that assemble into heterochromatin, a highly compacted structure that constrains their mutational potential. How heterochromatin forms during development and how its structure is maintained are not fully understood. Here, we show that mouse heterochromatin phase-separates after fertilization, during the earliest stages of mammalian embryogenesis. Using high-resolution quantitative imaging and molecular biology approaches, we show that pericentromeric heterochromatin displays properties consistent with a liquid-like state at the two-cell stage, which change at the four-cell stage, when chromocenters mature and heterochromatin becomes silent. Disrupting the condensates results in altered transcript levels of pericentromeric heterochromatin, suggesting a functional role for phase separation in heterochromatin function. Thus, our work shows that mouse heterochromatin forms membrane-less compartments with biophysical properties that change during development and provides new insights into the self-organization of chromatin domains during mammalian embryogenesis.

Trend of changes in epidermal biophysical properties in the Chinese aged 1 month to 17-year old.

Although it is known that epidermal biophysical properties vary with age and gender, the changes in epidermal biophysical properties over the time from baby to adolescence have not been elucidated yet. In the present study, we assessed the trend of changes in transepidermal water loss rates (TEWL), stratum corneum hydration, and skin surface pH in Chinese children. A total of 780 boys and 610 girls, aged 1 month to 17-year old, were enrolled in this study. TEWL and stratum corneum hydration on theforearmandtheshinwere measured with GPSkin Barrier, whereas skin surface pH was measured with portable skin pH meter. Overall, TEWL and stratum corneum hydration levels decreased, whereas skin surface pH increased in children from 1-month old to 17-year old. Significant decline in TEWL was observed on both the forearm and the shin of girls, and the shin of boys aged 13-17-year old. Similarly, marked decline in stratum corneum hydration levels started at ages of 6-12-year old. In contrast, decline in skin surface pH was observed in both girls and boys aged one to 12-month old except on the forearm of boys. Afterward, skin surface pH remained either stable or slight increase except on the shin of boys aged >12 months to 3-year old. These results demonstrate that both TEWL and stratum corneum hydration levels decrease, whereas skin surface pH increases in children aged 1 month to 17-year old. The changes in these biophysical properties vary with age, gender, and body site.

Tuning immunity through tissue mechanotransduction.

Immune responses are governed by signals from the tissue microenvironment, and in addition to biochemical signals, mechanical cues and forces arising from the tissue, its extracellular matrix and its constituent cells shape immune cell function. Indeed, changes in biophysical properties of tissue alter the mechanical signals experienced by cells in many disease conditions, in inflammatory states and in the context of ageing. These mechanical cues are converted into biochemical signals through the process of mechanotransduction, and multiple pathways of mechanotransduction have been identified in immune cells. Such pathways impact important cellular functions including cell activation, cytokine production, metabolism, proliferation and trafficking. Changes in tissue mechanics may also represent a new form of ‘danger signal’ that alerts the innate and adaptive immune systems to the possibility of injury or infection. Tissue mechanics can change temporally during an infection or inflammatory response, offering a novel layer of dynamic immune regulation. Here, we review the emerging field of mechanoimmunology, focusing on how mechanical cues at the scale of the tissue environment regulate immune cell behaviours to initiate, propagate and resolve the immune response.

Shape-Morphing of an Artificial Protein Cage with Unusual Geometry Induced by a Single Amino Acid Change.

Artificial protein cages are constructed from multiple protein subunits. The interaction between the subunits, notably the angle formed between them, controls the geometry of the resulting cage. Here, using the artificial protein cage, “TRAP-cage”, we show that a simple alteration in the position of a single amino acid responsible for Au(I)-mediated subunit–subunit interactions in the constituent ring-shaped building blocks results in a more acute dihedral angle between them. In turn, this causes a dramatic shift in the structure from a 24-ring cage with an octahedral symmetry to a 20-ring cage with a C2 symmetry. This symmetry change is accompanied by a decrease in the number of Au(I)-mediated bonds between cysteines and a concomitant change in biophysical properties of the cage.

Fingerprinting Small Molecule Modulators of Nucleolar Biophysics

Ribosomal RNA (rRNA) is the most abundant RNA in cells, reflecting its fundamental role in enabling protein synthesis in all living organisms. Nucleolus is a membraneless and multicompartmentalized nuclear body that governs rRNA biogenesis, a process that is misregulated in various diseases. Recent research has identified several small molecule therapeutics that alter rRNA biogenesis and subsequently nucleolar morphology, challenging the accepted mechanisms of action for these compounds and simultaneously establishing the nucleolus as an attractive therapeutic target. Nevertheless, there are currently no comprehensive and quantitative frameworks that capture and relate various drug‐induced changes in nucleolar morphology and its material properties to its function. We hypothesized that we can establish such a structure‐function framework by screening a panel of nucleolus‐perturbing small molecule drugs and measuring changes in biophysical properties of the nucleolus in cells from microscopy images. Here, we combine morphometric, thermodynamic, and kinetic measurements of drug‐treated nucleoli to reveal “fingerprints” for drugs that modulate distinct stages of rRNA biogenesis. We anticipate that this framework provides a starting point for robustly determining and predicting structure‐function relationships for nucleolar phenotype perturbations as well as for developing high‐content screening platforms to expedite the discovery of novel nucleolus‐targeting therapeutics.

Dual-frequency impedance assays for intracellular components in microalgal cells.

Intracellular components (including organelles and biomolecules) at the submicron level are typically analyzed in situ by special preparation or expensive setups. Here, a label-free and cost-effective approach of screening microalgal single-cells at a subcellular resolution is available based on impedance cytometry. To the best of our knowledge, it is the first time that the relationships between impedance signals and submicron intracellular organelles and biomolecules are shown. Experiments were performed on Euglena gracilis (E. gracilis) cells incubated under different incubation conditions (i.e., aerobic and anaerobic) and 15 μm polystyrene beads (reference) at two distinct stimulation frequencies (i.e., 500 kHz and 6 MHz). Based on the impedance detection of tens of thousands of samples at a throughput of about 900 cells per second, three metrics were used to track the changes in biophysical properties of samples. As a result, the electrical diameters of cells showed a clear shrinkage in cell volume and intracellular components, as observed under a microscope. The morphology metric of impedance pulses (i.e., tilt index) successfully characterized the changes in cell shape and intracellular composition distribution. Besides, the electrical opacity showed a stable ratio of the intracellular components to cell volume under the cellular self-regulation. Additionally, simulations were used to support these findings and to elucidate how submicron intracellular components and cell morphology affect impedance signals, providing a basis for future improvements. This work opens up a label-free and high-throughput way to analyze single-cell intracellular components by impedance cytometry.

Measurement of Incus and Stapes Velocity Following Sound Stimulus in the Setting of Incudostapedial Joint Separation and Reconstruction.

Understand the biophysical property changes of incudostapedial joint (ISJ) separation and ossicular hydroxyapatite application on middle ear function. Basic science. Cadaveric temporal bone research laboratory. A complete mastoidectomy was performed on five human temporal bones. A Laser Doppler Vibrometer was utilized to obtain velocity transfer function measurements of the incus and stapes across a range of frequencies in response to an acoustic stimulus. Under binocular microscopy the ISJ was separated and subsequently repaired with bone cement. Measurements were taken prior to ISJ separation, following joint separation, 30 to 60 min postrepair of the joint, and again 24 to 48 h postrepair of the joint. The stapes measurements taken from the intact ossicular chain and from the chains repaired with bone cement demonstrated a similar distribution of measurements. The ISJ separation showed dramatically reduced velocity transfer function stapes measurements but increased incus velocity transfer function measurements. In the early and delayed repaired chains, the mean velocity of the incus and stapes velocity peaked between 1.5 and 2 kHz, matching intact maximal velocity. Pure tone average at 0.5, 1, 2, and 3 kHz demonstrated no change in reconstructed stapes velocity at 24 to 48 h. Isolated ISJ separation in fresh frozen and thawed temporal bones produces stapes velocity transfer function changes that corresponds with the clinically experienced conductive hearing loss. Repair with bone cement produced similar velocity curves to the intact ISJ curve with excellent recovery across mid-frequencies. This model would be useful for future ossicular mechanical studies.

Changes in Biophysical Properties of Undifferentiated SH-SY5Y Cells During Long-term Cultures

The electrophysiological properties of undifferentiated SH-SY5Y cells were examined during cultures prolonged even to 20 days by measuring the passive and active membrane properties at 5 days interval, as well as the spontaneous spiking activity. The results showed that culturing this cell for long time affected not only membrane shape but also their electrophysiological properties. In particular, these cells considerably varied their sodium and potassium voltage-dependent currents, various channels kinetic features and their excitable properties. These modifications would synergically contribute to the bioelectrical conversion of these cells and could be part of a more complex machinery with which the tumoral cell would regulate its survival advantage and resilience. Understanding these processes could add a new clue to the exploitation of this preclinical human neuronal model.

Effect of Post-mortem Interval and Perfusion on the Biophysical Properties of ex vivo Liver Tissue Investigated Longitudinally by MRE and DWI.

Structural changes of soft tissues on the cellular level can be characterized by histopathology, but not longitudinally in the same tissue. Alterations of cellular structures and tissue matrix are associated with changes in biophysical properties which can be monitored longitudinally by quantitative diffusion-weighted imaging (DWI) and magnetic resonance elastography (MRE). In this work, DWI and MRE examinations were performed in a 0.5-Tesla compact scanner to investigate longitudinal changes in water diffusivity, stiffness and viscosity of ex-vivo rat livers for up to 20 h post-mortem (pm). The effect of blood on biophysical parameters was examined in 13 non-perfused livers (containing blood, NPLs) and 14 perfused livers (blood washed out, PLs). Changes in cell shape, cell packing and cell wall integrity were characterized histologically. In all acquisitions, NPLs presented with higher shear-wave speed (c), higher shear-wave penetration rate (a) and smaller apparent-diffusion-coefficients (ADCs) than PL. Time-resolved analysis revealed three distinct phases: (i) an initial phase (up to 2 h pm) with markedly increased c and a and reduced ADCs; (ii) an extended phase with relatively stable values; and (iii) a degradation phase characterized by significant increases in a (10 h pm in NPLs and PLs) and ADCs (10 h pm in NPLs, 13 h pm in PLs). Histology revealed changes in cell shape and packing along with decreased cell wall integrity, indicating tissue degradation in NPLs and PLs 10 h pm. Taken together, our results demonstrate that the biophysical properties of fresh liver tissue rapidly change within 2 h pm, which seems to be an effect of both cytotoxic edema and vascular blood content. Several hours later, disruption of cell walls resulted in higher water diffusivity and wave penetration. These results reveal the individual contributions of vascular components and cellular integrity to liver elastography and provide a biophysical, imaging-based fingerprint of liver tissue degradation.

Fano Resonance-Based Blood Plasma Monitoring and Sensing using Plasmonic Nanomatryoshka.

The fast label-free detection of specific antibodies and their concentration in blood plasma is useful for many applications, e.g., in Covid-19 patients. The change in biophysical properties like the refractive index of blood plasma due to the production of antibodies during infection may be very helpful in estimating the level and intensity of infection and subsequent treatment based on blood plasma therapy. In this article, Fano resonance-based refractive index sensor using plasmonic nanomatryoshka is proposed for blood plasma sensing. The interaction between hybridized modes (bright and dark modes) in optimized nanomatryoshka leads to Fano resonance, which by virtue of steeper dispersion can confine the light more efficiently compared with Lorentzian resonance. We propose the excitation of Fano resonances in sub 100-nm size nanomatryoshka based on newly emerging plasmonic materials ZrN and HfN, and one of the most widely used conventional plasmonic material, Au. Fano resonance-based plasmonic sensors leads to sensitivity = 188.5 nm/RIU, 242.5 nm/RIU, and 244.9 nm/RIU for Au, ZrN, and HfN, respectively. The corresponding figure of merit (nm/RIU) is ~ 3.5 times 103, 3.1 times 103, and 2.8 times 103 for Au, ZrN, and HfN, respectively. Present theoretical analysis shows that refractive index sensors with high sensitivity and figure of merit are feasible using Fano modes of plasmonic nanomatryoshka.

Biophysical and ultrasonographic changes in early patch/plaque stage of mycosis fungoides, compared with uninvolved skin

The goal of this study was evaluation of the skin biophysical properties in early patch/plaque stage of mycosis fungoides (MF) and its comparison with uninvolved skin in order to gain a better understanding of the pathogenesis of diseases. The stratum corneum hydration, transepidermal water loss (TEWL), surface friction, pH, sebum, melanin, erythema, temperature, elasticity parameters (R0, R2, R5), thickness, and echo density of epidermis and dermis were measured on lesions of 21 patients and compared with controls (average measures of uninvolved perilesional and symmetrical skins) by paired sample t test. Stratum corneum hydration (P<0.001) and echo density of dermis (P=0.044) were significantly lower, whereas pH (P-value=0.007), erythema (P<0.001), and melanin content (P=0.007) were significantly higher in lesions. There was not any significant difference in TEWL, friction index, sebum, temperature, R0, R2, R5, thickness of epidermis and dermis, and echo density of epidermis between lesions and normal skin. Parapsoriasis/MF lesions are specified by a set of certain changes in biophysical properties which are mainly correlated with histological changes. These sets of alterations may help in noninvasive, early diagnosis of parapsoriasis/MF.

Variations in the Biophysical Properties of DMPG and DPPG Membranes with Staphyloxanthin: A Molecular Dynamics Simulation Study

Staphyloxanthin (STX) is a carotenoid (glycolipid) that is part of the membrane of Staphylococcus aureus (S.aureus). It is related to antibiotic resistance because of the extended conjugated double bond chain that works as a free radical scavenger. in this study we explore the changes in biophysical properties of DMPG and DPPG membranes (two major components of S.aureus membranes) with 1 molecule and 15 mol% STX using molecular dynamics simulations with GROMACS and CHARMM36m force field. We performed quantum mechanical calculations (QM) to execute dihedral parameter optimization.

Cellular basis of ClC-2 Cl− channel–related brain and testis pathologies

The ClC-2 chloride channel is expressed in the plasma membrane of almost all mammalian cells. Mutations that cause the loss of ClC-2 function lead to retinal and testicular degeneration and leukodystrophy, whereas gain-of-function mutations cause hyperaldosteronism. Leukodystrophy is also observed with a loss of GlialCAM, a cell adhesion molecule that binds to ClC-2 in glia. GlialCAM changes the localization of ClC-2 and opens the channel by altering its gating. We now used cell type–specific deletion of ClC-2 in mice to show that retinal and testicular degeneration depend on a loss of ClC-2 in retinal pigment epithelial cells and Sertoli cells, respectively, whereas leukodystrophy was fully developed only when ClC-2 was disrupted in both astrocytes and oligodendrocytes. The leukodystrophy of Glialcam−/− mice could not be rescued by crosses with Clcn2op/op mice in which a mutation mimics the “opening” of ClC-2 by GlialCAM. These data indicate that GlialCAM-induced changes in biophysical properties of ClC-2 are irrelevant for GLIALCAM-related leukodystrophy. Taken together, our findings suggest that the pathology caused by Clcn2 disruption results from disturbed extracellular ion homeostasis and identifies the cells involved in this process.

A solution of identifying biophysical properties and 3D cellular structure of visible-light-driven photocatalytic inactivated Staphylococcus aureus

The mechanism of bacterial damage under visible-light-driven photo-inactivation system was attempted. Atomic force microscopy (AFM) and transmission X-ray microscopy (TXM) were firstly used to identify changes in biophysical properties, such as cellular height, roughness, adhesion, and modulus, and 3D cellular structure of microbial cells under visible-light-responsive N-TiO2 inactivation. Results revealed that the cell adhesion of N-TiO2 occurred immediately upon light irradiation then N-TiO2 particles penetrated the cell membrane, and deformed the cellular structure at the beginning of log phase under photocatalytic inactivation. The destruction of the outer cellular caused leakage of cellular K+ and lipid peroxidation, which ultimately brought about severe decrease in cell density. AFM and TXM provided direct observations on interactions between single cell and nano-materials at the nanoscale, which was useful for the early diagnosis of cell damage.

Recent land cover changes in the Southwestern US lead to an increase in surface temperature

The increase in large-scale land cover change (LCC) in recent decades, particularly in response to climate-driven disturbances, has potential to impact local and regional changes in climate due to modification of carbon sources and sinks, albedo, surface roughness and energy fluxes. Using observational data, we predict the impact of two of the most extensive LCCs occurring in the Southwestern US: drought-induced tree mortality and shrub encroachment into grasslands, on surface temperature. We developed a new energy balance method that extracts the biophysical responses to environmental conditions, to predict how structural changes in albedo, surface roughness and canopy conductance following LCC will alter surface temperature. This method allows us not only to explain the observed differences in surface temperature between two non-adjacent study sites with different environmental conditions, but also to separate the contribution of biophysical and non-biophysical properties to surface temperature. Our results suggest that changes in biophysical properties due to shrub encroachment and tree mortality in the Southwestern US (independent of changes in other environmental properties) can potentially lead to an increase in midday surface temperature (11AM to 2PM) of 1 to 2 degrees Celsius, comparable to changes in surface temperature following deforestation. Although the average surface temperature increase in response to both shrub encroachment and tree mortality is similar, the biophysical properties driving the temperature change are different in each scenario. Change in aerodynamic conductance following tree mortality is the largest contributor to heating, while reduced albedo and canopy conductance drive the increase in surface temperature following shrub encroachment. We also show how this increase in daytime surface temperature could be further intensified with future climate, especially with the expected reduction in soil water availability in the Southwest.

Anionic nanoparticle-induced perturbation to phospholipid membranes affects ion channel function

Understanding the mechanisms of nanoparticle interaction with cell membranes is essential for designing materials for applications such as bioimaging and drug delivery, as well as for assessing engineered nanomaterial safety. Much attention has focused on nanoparticles that bind strongly to biological membranes or induce membrane damage, leading to adverse impacts on cells. More subtle effects on membrane function mediated via changes in biophysical properties of the phospholipid bilayer have received little study. Here, we combine electrophysiology measurements, infrared spectroscopy, and molecular dynamics simulations to obtain insight into a mode of nanoparticle-mediated modulation of membrane protein function that was previously only hinted at in prior work. Electrophysiology measurements on gramicidin A (gA) ion channels embedded in planar suspended lipid bilayers demonstrate that anionic gold nanoparticles (AuNPs) reduce channel activity and extend channel lifetimes without disrupting membrane integrity, in a manner consistent with changes in membrane mechanical properties. Vibrational spectroscopy indicates that AuNP interaction with the bilayer does not perturb the conformation of membrane-embedded gA. Molecular dynamics simulations reinforce the experimental findings, showing that anionic AuNPs do not directly interact with embedded gA channels but perturb the local properties of lipid bilayers. Our results are most consistent with a mechanism in which anionic AuNPs disrupt ion channel function in an indirect manner by altering the mechanical properties of the surrounding bilayer. Alteration of membrane mechanical properties represents a potentially important mechanism by which nanoparticles induce biological effects, as the function of many embedded membrane proteins depends on phospholipid bilayer biophysical properties.

Proteomic, biomechanical and functional analyses define neutrophil heterogeneity in systemic lupus erythematosus

ObjectivesLow-density granulocytes (LDGs) are a distinct subset of proinflammatory and vasculopathic neutrophils expanded in systemic lupus erythematosus (SLE). Neutrophil trafficking and immune function are intimately linked to cellular biophysical properties....

88. Cell stiffness decreases with severity of disc degeneration and inflammatory stimulation

88. Cell stiffness decreases with severity of disc degeneration and inflammatory stimulation

Abstract P3-02-11: Topographical ues overlaid with fructose-like molecules to assess breast cancer recruitment in nanofiber scaffolds

Abstract Tumorigenesis is a complex process involving numerous cellular signaling cascades and environmental factors. While 2D cultures for stiffness measurements and more recently 3D cultures to demonstrate differences in cell structures have been reported, very little is known about the impact of sugars in cell recruitment. In this study, we report the fabrication of 3D-scaffolds with different morphologies obtained by electrospinning with fluorescent fructose-like molecular probes to study cancer cell proliferation and migration. Using a FDA approved, biocompatible and biodegradable polymer Polycaprolactone (PCL), we electrospun nanofiber scaffolds having random, aligned, and honeycomb morphologies. Scaffolds with similar morphology without the fructose mimicking analogs were fabricated and used as controls. The degradation rate of the scaffolds was also characterized to ensure long-term availability of probes and mechanical stability of PCL. Cell viability, cell morphology, formation of colonies with changes in morphology were investigated. The changes in biophysical properties of tumor microenvironment with change in morphology of the scaffolds were investigated. In vitro tests for proliferation, alignment and migration of normal breast epithelial cells (184B5), adenocarcinoma (MCF-7), pre-malignant (MCF10AneoT) and triple-negative (MDAMB231 on the scaffolds on days 1, 2, and 3 were carried out. The morphology of the scaffolds was characterized using FE-SEM; cell proliferation was assessed using CellTiter-Blue® Viability Assay; migration and cell-scaffold interactions were investigated using phalloidin for F-Actin and fructose response was determined by immunostaining for facilitative fructose transporter GLUT-5. Our data indicates that while topographical features affected cell adhesion and proliferation, cell lines that responded to the fructose-like probes tended to be more invasive. Furthermore, the preference to a specific scaffold was greatly altered by the presence of the probes with MDAMB231 showing least preference after 72 hours and pre-malignant AneoT showing highest preference. However, there was no significant difference in the cell numbers between scaffolds with probes and those without for the pre-malignant cells while this difference was noticeable in the control cell lines. Further investigation into specific response to glucose and fructose uptake through their major transporters GLUT2 and GLUT5 are currently ongoing. Citation Format: Carolynn Que, Samerender Nagam Hanumantharao, Brennan Vogl, Marina Tanasova, Smitha Rao. Topographical ues overlaid with fructose-like molecules to assess breast cancer recruitment in nanofiber scaffolds [abstract]. In: Proceedings of the 2019 San Antonio Breast Cancer Symposium; 2019 Dec 10-14; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2020;80(4 Suppl):Abstract nr P3-02-11.