Search

Fluorogenic probes that unmask fluorescence signals in response to bioorthogonal reactions are a powerful new addition to biological imaging. They can significantly reduce background fluorescence and minimize nonspecific signals, potentially enabling real-time, high-contrast imaging without the need to wash out excess fluorophores. While diverse classes of highly refined synthetic fluorophores are now readily available, integrating them into a bioorthogonal fluorogenic scheme still requires extensive design efforts and customized structural alterations to optimize quenching mechanisms for each specific fluorophore scaffold. Herein, we present a highly generalizable strategy that can produce an efficient bioorthogonal fluorogenic response from essentially any readily available fluorophore without further structural alterations. We designed this strategy based on the macrocyclic cucurbit[7]uril (CB7) host, where a fluorogenic response is achieved by programming a guest exchange reaction within the macrocyclic cavity. We employed this strategy to rapidly create fluorogenic probes across the visible spectrum from diverse fluorophore scaffolds, which enabled no-wash imaging in live cells and tissues with minimal background signal. Finally, we demonstrated that this strategy can be combined with metabolic labeling for fluorogenic detection of metabolically tagged mycobacteria under no-wash conditions and paired with covalently clickable probes for high-contrast super-resolution and multiplexed imaging in cells and tissues., Competing Interests: The authors declare no competing financial interest., (© 2024 The Authors. Published by American Chemical Society.)

Loss of retinal ganglion cells (RGCs) is central to the pathogenesis of optic neuropathies such as glaucoma. Increased RGC cAMP signaling is neuroprotective. We have shown that displacement of the cAMP-specific phosphodiesterase PDE4D3 from an RGC perinuclear compartment by expression of the modified PDE4D3 N-terminal peptide 4D3(E) increases perinuclear cAMP and protein kinase A activity in cultured neurons and in vivo RGC survival after optic nerve crush (ONC) injury. To explore mechanisms by which PDE4D3 displacement promotes neuroprotection, in this study mice intravitreally injected with an adeno-associated virus to express an mCherry-tagged 4D3(E) peptide were subjected to ONC injury and analyzed by single cell RNA-sequencing (scRNA-seq). 4D3(E)-mCherry expression was associated with an attenuation of injury-induced changes in gene expression, thereby supporting the hypothesis that enhanced perinuclear PKA signaling promotes neuroprotective RGC gene expression., Competing Interests: Declaration of competing interest J.L.G. and M.S.K. are the inventors for US patent application 17/290,174 concerning PDE4D3 anchoring disruption in neuroprotection., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Skin serves as both barrier and interface between body and environment. Skin microbes are intermediaries evolved to respond, transduce, or act in response to changing environmental or physiological conditions. We quantified genome-wide changes in gene expression levels for one abundant skin commensal, Staphylococcus epidermidis , in response to an internal physiological signal, glucose levels, and an external environmental signal, temperature. We found 85 of 2,354 genes change up to ~ 34-fold in response to medically relevant changes in glucose concentration (0-17 mM; adj p ≤0.05). We observed carbon catabolite repression in response to a range of glucose spikes, as well as upregulation of genes involved in glucose utilization in response to persistent glucose. We observed 366 differentially expressed genes in response to a physiologically relevant change in temperature (37-45°C; adj p ≤ 0.05) and an S. epidermidis heat-shock response that mostly resembles the heat-shock response of related staphylococcal species. DNA motif analysis revealed CtsR and CIRCE operator sequences arranged in tandem upstream of dnaK and groESL operons. We identified and curated 38 glucose-responsive genes as candidate ON or OFF switches for use in controlling synthetic genetic systems. Such systems might be used to instrument the in-situ skin microbiome or help control microbes bioengineered to serve as embedded diagnostics, monitoring, or treatment platforms., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Benjamin, Goyal, Nair and Endy.)

Microglia are implicated as primarily detrimental in pain models; however, they exist across a continuum of states that contribute to homeostasis or pathology depending on timing and context. To clarify the specific contribution of microglia to pain progression, we take advantage of a temporally controlled transgenic approach to transiently deplete microglia. Unexpectedly, we observe complete resolution of pain coinciding with microglial repopulation rather than depletion. We find that repopulated mouse spinal cord microglia are morphologically distinct from control microglia and exhibit a unique transcriptome. Repopulated microglia from males and females express overlapping networks of genes related to phagocytosis and response to stress. We intersect the identified mouse genes with a single-nuclei microglial dataset from human spinal cord to identify human-relevant genes that may ultimately promote pain resolution after injury. This work presents a comprehensive approach to gene discovery in pain and provides datasets for the development of future microglial-targeted therapeutics., Competing Interests: Declaration of interests The authors declare that they have no competing interests., (Copyright © 2024 The Author(s). Published by Elsevier Inc. All rights reserved.)

Color centers in nanodiamonds (NDs) have been largely explored by coupling to a photonic structured matrix (PSM) to amplify visible range emission features, enhancing their use in quantum technologies. Here, we study the emission enhancement of dual near-infrared zero phonon line (ZPL) emission from silicon-boron (SiB) and silicon-vacancy (SiV - ) centers in NDs using a spontaneously emerged low index-contrast quasiperiodic PSM, having micron-scale air pores. An intensity enhancement factor of 6.15 for SiV - and 7.8 for SiB ZPLs is attained for the PSM sample compared to a control sample. We find Purcell enhancement of 2.77 times for the PSM sample using spatial-dependent decay rate measurements, supported by localized field intensity confinement in the sample. Such cavity-like emission enhancement and lifetime reduction are enabled by an in-plane order-disorder scattering in the PSM sample substantiated by pump-dependent emission measurements. The results put forward a facile approach to tailor the near-infrared dual ZPL emission from NDs using nanophotonic structures.

Polymorphic forms of organic conjugated small molecules, with their unique molecular shapes, packing arrangements, and interaction patterns, provide an excellent opportunity to uncover how their microstructures influence their observable properties. Ethyl-2-(1-benzothiophene-2-yl)quinoline-4-carboxylate (BZQ) exists as dimorphs with distinct crystal habits - blocks (BZB) and needles (BZN). The crystal forms differ in their molecular arrangements - BZB has a slip-stacked column-like structure in contrast to a zig-zag crystal packing with limited π-overlap in BZN. The BZB crystals characterized by extended π-stacking along [100] demonstrated semiconductor behavior, whereas the BZN, with its zig-zag crystal packing and limited stacking characteristics, was reckoned as an insulator. Monotropically related crystal forms also differ in their nanomechanical properties, with BZB crystals being considerably softer than BZN crystals. This discrepancy in mechanical behavior can be attributed to the distinct molecular arrangements adopted by each crystal form, resulting in unique mechanisms to relieve the strain generated during nanoindentation experiments. Waveguiding experiments on the acicular crystals of BZN revealed the passive waveguiding properties. Excitation of these crystals using a 532 nm laser confirmed the propagation of elastically scattered photons (green) and the subsequent generation of inelastically scattered (orange) photons by the crystals. Further, the dimorphs display dissimilar photoluminescence properties; they are both blue-emissive, but BZN displays twice the quantum yield of BZB. The study underscores the integral role of polymorphism in modulating the mechanical, photophysical, and conducting properties of functional molecular materials. Importantly, our findings reveal the existence of light-emitting crystal polymorphs with varying electric conductivity, a relatively scarce phenomenon in the literature., (© 2023 Wiley-VCH GmbH.)

The intrinsic mechanisms that regulate neurotoxic versus neuroprotective astrocyte phenotypes and their effects on central nervous system degeneration and repair remain poorly understood. Here we show that injured white matter astrocytes differentiate into two distinct C3-positive and C3-negative reactive populations, previously simplified as neurotoxic (A1) and neuroprotective (A2) 1,2 , which can be further subdivided into unique subpopulations defined by proliferation and differential gene expression signatures. We find the balance of neurotoxic versus neuroprotective astrocytes is regulated by discrete pools of compartmented cyclic adenosine monophosphate derived from soluble adenylyl cyclase and show that proliferating neuroprotective astrocytes inhibit microglial activation and downstream neurotoxic astrocyte differentiation to promote retinal ganglion cell survival. Finally, we report a new, therapeutically tractable viral vector to specifically target optic nerve head astrocytes and show that raising nuclear or depleting cytoplasmic cyclic AMP in reactive astrocytes inhibits deleterious microglial or macrophage cell activation and promotes retinal ganglion cell survival after optic nerve injury. Thus, soluble adenylyl cyclase and compartmented, nuclear- and cytoplasmic-localized cyclic adenosine monophosphate in reactive astrocytes act as a molecular switch for neuroprotective astrocyte reactivity that can be targeted to inhibit microglial activation and neurotoxic astrocyte differentiation to therapeutic effect. These data expand on and define new reactive astrocyte subtypes and represent a step towards the development of gliotherapeutics for the treatment of glaucoma and other optic neuropathies., (© 2023. The Author(s), under exclusive licence to Springer Nature Limited.)

Pulmonary arterial hypertension (PAH) is a progressive disease in which pulmonary arterial (PA) endothelial cell (EC) dysfunction is associated with unrepaired DNA damage. BMPR2 is the most common genetic cause of PAH. We report that human PAEC with reduced BMPR2 have persistent DNA damage in room air after hypoxia (reoxygenation), as do mice with EC-specific deletion of Bmpr2 (EC-Bmpr2 -/- ) and persistent pulmonary hypertension. Similar findings are observed in PAEC with loss of the DNA damage sensor ATM, and in mice with Atm deleted in EC (EC-Atm -/- ). Gene expression analysis of EC-Atm -/- and EC-Bmpr2 -/- lung EC reveals reduced Foxf1, a transcription factor with selectivity for lung EC. Reducing FOXF1 in control PAEC induces DNA damage and impaired angiogenesis whereas transfection of FOXF1 in PAH PAEC repairs DNA damage and restores angiogenesis. Lung EC targeted delivery of Foxf1 to reoxygenated EC-Bmpr2 -/- mice repairs DNA damage, induces angiogenesis and reverses pulmonary hypertension., (© 2023. The Author(s).)

Ultrasmall metal nanoclusters are attractive for their size-dependent optical and electrochemical properties. Here, blue-emitting copper clusters stabilized with cetyltrimethylammonium bromide (CTAB) are synthesized using an electrochemical approach. The electrospray ionization (ESI) analysis reveals that the cluster has 13 copper atoms in the core. The clusters are then used for the electrochemical detection of endotoxin─a bacterial toxin in Gram-negative bacteria. Differential pulse voltammetry (DPV) is used to detect endotoxins with high selectivity and sensitivity. It shows a lower detection limit of 100 ag mL -1 with a linear range of 100 ag mL -1 to 10 ng mL -1 . The sensor is efficient for detecting endotoxins from human blood serum samples.

Bacterial contamination is a serious concern for health and environmental safety. The major toxic effect arises from the endotoxin or lipopolysaccharide (LPS) attached to the cell wall of the gram-negative bacteria. Ultrasensitive endotoxin detection is of supreme importance in sustaining food, clinical and pharmaceutical safety. Herein we report a simple electrochemical detection platform using reduced graphene oxide (rGO) combined with cuprous oxide nanoparticles for the ultrasensitive detection of LPS. The sensor uses polymyxin B (PmB) to achieve the selective response towards LPS. The sensor showed a lower detection limit (LOD) of 10 agmL -1 with linearity from 10 agmL -1 to 10 ngmL -1 . Detection of LPS from whole blood is also carried out with excellent sensitivity. The sensor showed excellent recovery rates in whole blood, pointing to the capability of using the sensor in real-life clinical analysis. The sensor detects Gram-negative bacteria from sewage water with a rapid response time, indicating the effectiveness of the sensor in water quality analysis., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Background: Effective cancer treatment relies on precision diagnostics. In cytology, an accurate diagnosis facilitates the determination of proper therapeutics for patients with cancer. Previously, the authors developed a multiplexed immunofluorescent panel to detect epithelial malignancies from pleural effusion specimens. Their assay reliably distinguished effusion tumor cells (ETCs) from nonmalignant cells; however, it lacked the capacity to reveal specific cancer origin information. Furthermore, DNA profiling of ETCs revealed some, but not all, cancer-driver mutations., Methods: The authors developed a new multiplex immunofluorescent panel that detected both malignancy and pulmonary origin by incorporating the thyroid transcription factor-1 (TTF-1) biomarker. Evaluation for TTF-1-positive ETCs (T-ETCs) was performed on 12 patient samples. T-ETCs and parallel ETCs from selected patients were collected and subjected to DNA profiling to identify pathogenic mutations. All samples were obtained with Institutional Review Board approval., Results: Malignancy was detected in all samples. T-ETCs were identified in 9 of 10 patients who had clinically reported TTF-1 positivity (90% sensitivity and 100% specificity). Furthermore, DNA profiling of as few as five T-ETCs identified pathogenic mutations with equal or greater sensitivity compared with profiling of ETCs, both of which showed high concordance with clinical findings., Conclusions: The findings suggest that the immunofluorescent and molecular characterization of tumor cells from pleural effusion specimens can provide reliable diagnostic information, even with very few cells. The integration of site-specific biomarkers like TTF-1 into ETC analysis may facilitate better refined diagnosis and improve patient care., (© 2022 American Cancer Society.)

Engineering different nanomaterials into a single functional material can impart unique properties of the parental nanoparticles, especially in the field of bio imaging and therapy. Gold nanomaterials having different sizes, shapes and dimensionalities exhibit exceptional properties apart from their non-toxicity and hence are strong candidates in the biomedical field. Designing a hybrid nanomaterial of two gold nanostructures retaining the individual properties of the parental nanomaterials is challenging. Here, we demonstrate the synthesis of a hybrid nanomaterial (GQC@GNR), comprising an extremely small gold nanocluster and a representative of the asymmetric gold nanostructure, i.e. , a gold nanorod, both having their own different exclusive optical properties like tuneable emission and NIR absorption characteristics, respectively. The hybrid system is designed to retain its emission and absorption in the NIR region to use it as an agent for simultaneous imaging and therapy. The formation of GQC@GNR and its architectonics heavily depend on the synthesis route and the parameters adopted which in turn have a direct influence on its properties. The architecture and its connection to the optical properties are explained using UV-Vis absorption and photoluminescence spectroscopy, zeta potential, transmission electron microscopy, etc . DFT-based computational modelling supports architectonics as explained by the experimental findings. The formation of the gold-gold hybrid system witnessed interesting science with a strong indication that materials with desired properties can be designed by appropriately modulating the architectonics of hybrid formation. Finally, folate conjugated GQC@GNR demonstrated its efficacy for targeted imaging and photothermal therapy in HeLa cells and tumor-bearing animal models. The detailed therapeutic efficacy of GQC@GNR is also explained based on Raman spectroscopy.

Endotoxins or lipopolysaccharides (LPS) present in the outer layer of Gram-negative bacteria (GNB) are responsible for bacterial toxicity. It is an environmental hazard that everyone is exposed to daily to various extents. Due to its potent toxicity, quantitative detection with very high sensitivity is essential in the food, medical, and pharmaceutical industries. Herein, we report an optical nanosensor for the rapid and sensitive detection of LPS and GNB based on the Cu 2+ -mediated aggregation of gold nanoparticles (Cu@AuNPs). The sensor detects LPS within a linear range of 20 ag/mL to 20 ng/mL with a lower detection limit of 0.2 ag/mL. The sensor could successfully recover spiked endotoxin in grape juice with a percentage error of ±0.2, confirming its application in the food industry. The sensor could also distinguish Gram-negative bacteria from Gram-positive bacteria, and the selectivity of the Cu@AuNP sensor toward GNB is utilized to detect Escherichia coli in wastewater. The rapid detection of E. coli without any pretreatment is a promising strategy in water analysis.

Physiologic laminar shear stress (LSS) induces an endothelial gene expression profile that is vasculo-protective. In this report, we delineate how LSS mediates changes in the epigenetic landscape to promote this beneficial response. We show that under LSS, KLF4 interacts with the SWI/SNF nucleosome remodeling complex to increase accessibility at enhancer sites that promote the expression of homeostatic endothelial genes. By combining molecular and computational approaches we discover enhancers that loop to promoters of KLF4- and LSS-responsive genes that stabilize endothelial cells and suppress inflammation, such as BMPR2, SMAD5, and DUSP5. By linking enhancers to genes that they regulate under physiologic LSS, our work establishes a foundation for interpreting how non-coding DNA variants in these regions might disrupt protective gene expression to influence vascular disease., (© 2022. The Author(s).)

The advent of photonic crystals has made possible the idea of controlling flows of light, which has revolutionized photonics-based technology. Photonic crystals are constructed based on periodic refractive index variations in one-, two-, or three-spatial dimensions on the optical-wavelength scale. Photonic crystals show inherent photonic stop gaps or band gaps depending upon the crystal symmetry and refractive index contrast. Showing ease of fabrication, polymer-based self-assembled photonic crystals with stop gaps have been widely explored. We discuss angle- and polarization-dependent stop gap creation and splitting at higher angles of incidence. The observed stop gaps in self-assembled photonic crystals often deviate from theoretical predictions due to experimental constraints, such as finite size and fabrication disorders associated with samples. We perform micro-reflectivity experiments on a single domain, showing minimal disorder, with nearly 100% reflectivity, which is in agreement with theory. We obtain more than 75% emission intensity suppression and a 30% increase in the emission lifetime at the stop gap using micro-emission experiments in a single domain. This enables us to study the role of finite-size effects in photonic crystals in modifying the emission properties. We observe the linear scaling of the emission intensity suppression and the emission rate with the finite size of the crystal. Our single-domain experimental studies reveal that the use of low index-contrast self-assembled photonic crystals is a potential platform for strategically modifying light transport and emission properties.

Development of selective, ultra-sensitive, rapid and facile methods for the detection of chemical residues of toxic pesticides and hazardous chemicals are quite important in food safety, environmental monitoring and safeguarding public health. Herein, we presented a fluorescent turn-on aptasensor based on sulphur-doped graphene quantum dot (S-GQD) utilizing specific recognition and binding property of aptamer for the ultra-sensitive and selective detection of omethoate (OM) which is a systemic organophosphorus pesticide. The detection method is based on tuning aggregation-disaggregation mechanism of S-GQD by way of conformational alteration of the recognition probe. Fluorescence 'turn-on' process includes aggregation-induced quenching of S-GQD with aptamer via S-GQD-aptamer complex formation and its subsequent fluorescence recovery with the addition of OM by structural switching of S-GQD-aptamer complex to aptamer-omethoate complex. The reported 'switch-on' aptasensor has exhibited a low limit of detection of 0.001 ppm with high selectivity for OM over other pesticides., Competing Interests: Declaration of competing interest None., (Copyright © 2021 Elsevier B.V. All rights reserved.)

The application of growth factor based therapies in regenerative medicine is limited by the high cost, fast degradation kinetics, and the multiple functions of these molecules in the cell, which requires regulated delivery to minimize side effects. Here a photoactivatable peptidomimetic of the vascular endothelial growth factor (VEGF) that allows the light-controlled presentation of angiogenic signals to endothelial cells embedded in hydrogel matrices is presented. A photoresponsive analog of the 15-mer peptidomimetic Ac-KLTWQELYQLKYKGI-NH 2 (abbreviated P QK) is prepared by introducing a 3-(4,5-dimethoxy-2-nitrophenyl)-2-butyl (DMNPB) photoremovable protecting group at the Trp4 residue. This modification inhibits the angiogenic potential of the peptide temporally. Light exposure of P QK modified hydrogels provide instructive cues to embedded endothelial cells and promote angiogenesis at the illuminated sites of the 3D culture, with the possibility of spatial control. P QK modified photoresponsive biomaterials offer an attractive approach for the dosed delivery and spatial control of pro-angiogenic factors to support regulated vascular growth by just using light as an external trigger., (© 2021 The Authors. Advanced Healthcare Materials published by Wiley-VCH GmbH.)

Background In complex congenital heart disease patients such as those with tetralogy of Fallot, the right ventricle (RV) is subject to pressure overload, leading to RV hypertrophy and eventually RV failure. The mechanisms that promote the transition from stable RV hypertrophy to RV failure are unknown. We evaluated the role of mitochondrial bioenergetics in the development of RV failure. Methods and Results We created a murine model of RV pressure overload by pulmonary artery banding and compared with sham-operated controls. Gene expression by RNA-sequencing, oxidative stress, mitochondrial respiration, dynamics, and structure were assessed in pressure overload-induced RV failure. RV failure was characterized by decreased expression of electron transport chain genes and mitochondrial antioxidant genes (aldehyde dehydrogenase 2 and superoxide dismutase 2) and increased expression of oxidant stress markers (heme oxygenase, 4-hydroxynonenal). The activities of all electron transport chain complexes decreased with RV hypertrophy and further with RV failure (oxidative phosphorylation: sham 552.3±43.07 versus RV hypertrophy 334.3±30.65 versus RV failure 165.4±36.72 pmol/(s×mL), P <0.0001). Mitochondrial fission protein DRP1 (dynamin 1-like) trended toward an increase, while MFF (mitochondrial fission factor) decreased and fusion protein OPA1 (mitochondrial dynamin like GTPase) decreased. In contrast, transcription of electron transport chain genes increased in the left ventricle of RV failure. Conclusions Pressure overload-induced RV failure is characterized by decreased transcription and activity of electron transport chain complexes and increased oxidative stress which are associated with decreased energy generation. An improved understanding of the complex processes of energy generation could aid in developing novel therapies to mitigate mitochondrial dysfunction and delay the onset of RV failure.

Precise control of light is indispensable to modern optical communication devices especially as the size of such devices approaches the subwavelength scale. Plasmonic devices are suitable for the development of these optical devices due to the extreme field confinement and its ability to be controlled by tuning the carrier density at the metal/dielectric interface. Here, an electro-ionic controlled plasmonic device consisting of Au/graphene/ion-gel is demonstrated as an optical switch, where an external electric field modulates the real part of the electrical conductivity. The graphene layer enhances charge penetration and charge separation at the Au/graphene interface resulting in an increased photoinduced voltage. The ion-gel immobilized on the Au/graphene further enables the electrical tunability of plasmons which modulates the intensity of the reflected laser light. This work paves the way for developing novel plasmonic electro-optic switches for potential applications such as integrated optical devices.

The design of molecular compounds that exhibit flexibility is an emerging area of research. Although a fair amount of success has been achieved in the design of plastic or elastic crystals, realizing multidimensional plastic and elastic bending remains challenging. We report herein a naphthalidenimine-boron complex that showed size-dependent dual mechanical bending behavior whereas its parent Schiff base was brittle. Detailed crystallographic and spectroscopic analysis revealed the importance of boron in imparting the interesting mechanical properties. Furthermore, the luminescence of the molecule was turned-on subsequent to boron complexation, thereby allowing it to be explored for multimode optical waveguide applications. Our in-depth study of the size-dependent plastic and elastic bending of the crystals thus provides important insights in molecular engineering and could act as a platform for the development of future smart flexible materials for optoelectronic applications., (© 2020 Wiley-VCH GmbH.)

The COVID-19 pandemic caused by the SARS-CoV-2 virus, has changed the homeostasis of the medical world. In this critical phase, in addition to the general recommendations issued by World Health Organization (WHO) for medical practitioners and health care givers, certain other precautions and safe care practices need to be emphasized which are unique to each branch of medicine. Aesthetic dermatology is no exception. With aesthetic treatments on the rise, it is pertinent to formulate safe practices for aesthetic dermatology to protect the doctor, health staff and the patients from getting exposed during this phase and in the aftermath of the pandemic. Recommendations for surgical and dental procedures advice to defer such procedures. This can be extrapolated to aesthetic dermatology also, but once health care services start, there should be some safety recommendations to be followed until we have definitive management or a vaccine for it., Competing Interests: There are no conflicts of interest., (Copyright: © 2020 Indian Dermatology Online Journal.)

Direct visualization of soft organic molecules like cellulose is extremely challenging under a high-energy electron beam. Herein, we adopt two ionization damage extenuation strategies to visualize the lattice arrangements of the β-(1→4)-d-glucan chains in carboxylated nanocellulose fibers (C-NCFs) having cellulose II crystalline phase using high-resolution transmission electron microscopy. Direct imaging of individual nanocellulose fibrils with high-resolution and least damage under high-energy electron beam is achieved by employing reduced graphene oxide, a conducting material with high electron transmittance and Ag + ions, with high electron density, eliminating the use of sample-specific, toxic staining agents, or other advanced add-on techniques. Furthermore, the imaging of cellulose lattices in a C-NCF/TiO 2 nanohybrid system is accomplished in the presence of Ag + ions in a medium revealing the mode of association of C-NCFs in the system, which validates the feasibility of the presented strategy. The methods adopted here can provide further understanding of the fine structures of carboxylated nanocellulose fibrils for studying their structure-property relationship for various applications., Competing Interests: The authors declare no competing financial interest., (Copyright © 2020 American Chemical Society.)

Controlling bacterial growth using artificial nanostructures inspired from natural species is of immense importance in biomedical applications. In the present work, a low cost, fast processing, and scalable anisotropic wet etching technique is developed to fabricate the densely packed disordered silicon nanopyramids (SiNPs) with nanosized sharp tips. The bactericidal characteristics of SiNPs are assessed against strains implicated in nosocomial and biomaterial-related infections. Compared to the bare silicon with no antibacterial activities, SiNPs of 1.85 ± 0.28 μm height show 55 and 75% inhibition of Escherichia coli (Gram-negative) and Bacillus subtilis (Gram-positive) bacteria, whereas the silicon nanowires (SiNWs) fabricated using a metal-assisted chemical etching method show 50 and 58% inhibition of E. coli and B. subtilis . The mechanistic studies using a scanning electron microscope and live/dead bacterial cell assay reveal cell rupture and predominance of dead cells on contact with SiNPs and SiNWs, which confirms their bactericidal effects. Chemical stability and cell viability studies demonstrate the biocompatible nature of SiNP and SiNW surfaces. Owing to their capability to kill both Gram-negative and positive bacteria and minimal toxicity to murine fibroblast cells, SiNPs can be used as an antibacterial coating on medical devices to prevent nosocomial and biomaterial-related infections.

Background: Tumors comprise a complex microenvironment of interacting malignant and stromal cell types. Much of our understanding of the tumor microenvironment comes from in vitro studies isolating the interactions between malignant cells and a single stromal cell type, often along a single pathway., Result: To develop a deeper understanding of the interactions between cells within human lung tumors, we perform RNA-seq profiling of flow-sorted malignant cells, endothelial cells, immune cells, fibroblasts, and bulk cells from freshly resected human primary non-small-cell lung tumors. We map the cell-specific differential expression of prognostically associated secreted factors and cell surface genes, and computationally reconstruct cross-talk between these cell types to generate a novel resource called the Lung Tumor Microenvironment Interactome (LTMI). Using this resource, we identify and validate a prognostically unfavorable influence of Gremlin-1 production by fibroblasts on proliferation of malignant lung adenocarcinoma cells. We also find a prognostically favorable association between infiltration of mast cells and less aggressive tumor cell behavior., Conclusion: These results illustrate the utility of the LTMI as a resource for generating hypotheses concerning tumor-microenvironment interactions that may have prognostic and therapeutic relevance.

The transition of conventional medicine to personalized medicine has paved the way for sensing new biomolecules. Consequently, this field attracted wide interest due to its capability to provide information on point of care basis. Multi-analyte sensors that emerged recently can perform quick and affordable analysis with minimum quantity of blood samples compared to traditional sensing of individual analytes. The present study focuses on the development of a quantum dot (Qd) based nanosensor for the simultaneous detection of copper and creatinine; two biologically relevant molecules. The sensor was designed by forming a complex of Qd with 1-Ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC) and picric acid through carboxylic bond formation of Qd-EDC with picric acid. The dual independent emissions of the Qd-EDC complex was used for the simultaneous detection of creatinine and copper by a turn on/turn off method and was successfully demonstrated with a sensitivity of nanomolar to millimolar, and micromolar to millimolar range respectively. The multianalyte sensor thus developed has quick response and works well under normal conditions of temperature and pH. It is also shown to work in cellular environment and blood serum. A simple image based detection of creatinine using the sensor strips has also been attempted by means of a mobile camera and validated with human blood samples., Competing Interests: Declaration of competing interest None, (Copyright © 2019 Elsevier B.V. All rights reserved.)

Early stage detection of neurodegenerative diseases such as Alzheimer's disease (AD) is of utmost importance, as it has become one of the leading causes of death of millions of people. The gradual intellectual decline in AD patients is an outcome of fibrillation of amyloid beta 1-42 (Aβ 1-42 ) peptides in the brain. In this paper, we present localized surface plasmon resonance (LSPR) based sensing of Aβ 1-42 fibrillation using Au nano-urchins. Strongly localized field confinement at the spiky nanostructures of nano-urchin surfaces enables them to detect very low concentrations of Aβ 1-42 . In addition, the LSPR peak of Au nano-urchins, which is very sensitive to ambient conditions, shows significant responses at different fibrillation stages of Aβ 1-42 . Reduction in LSPR peak intensity with an increase in the fibrillation is chosen as the sensing parameter here. This paper in this context provides LSPR based highly sensitive, label-free and real-time sensing of Aβ 1-42 fibrillation that is highly advantageous compared to the existing techniques which require binding additives or fluorescent biomarkers., Competing Interests: There are no conflicts to declare., (This journal is © The Royal Society of Chemistry.)

Near Infra-Red Surface Enhanced Raman Spectroscopy (NIR SERS) has gained huge attention in recent years as the conventional visible SERS suffers from overwhelming fluorescence background from the fluorophore resulting in the masking of Raman signals. In this paper, we propose a novel multi-layered SERS substrate- (Cu 2 O - Au) - Graphene - Au - for efficient NIR SERS applications. The proposed structure has a monolayer of Cu 2 O - Au core-shell particles on a Au substrate with 1 nm thick graphene spacer layer. Mie simulations are used to optimize the aspect ratios of core-shell particles to shift their plasmon resonances to NIR region using MieLab software. Further, Finite Difference Time Domain (FDTD) simulations using Lumerical software are used for the design of the multiparticle layered SERS substrate as MieLab software works only for single particle systems. Designed structure is shown to provide high field enhancement factor of the order of 10 8 at an excitation of 1064 nm thus ensuring the possibility of using the proposed structure as efficient NIR SERS substrate which could probably be used for various NIR sensing applications.

Strong plasmon absorption in the near-infrared (NIR) region renders gold nanorods (GNRs) amenable for biomedical applications, particularly for photothermal therapy. However, these nanostructures have not been explored for their imaging potential because of their weak emission profile. In this study, the weak fluorescence emission of GNRs is tuned to match that of the absorption of a photosensitizer (PS) molecule, and energy transfer from the GNR to PS enhances the emission profile of the GNR-PS combination. GNR complexes generally quench the fluorescence emission of nearby chromophores. However, herein, the complex retains or rather enhances the fluorescence through competition in energy transfer. Excitation-dependent energy transfer has been explained experimentally and theoretically by using DFT calculations, the CIE chromaticity diagram, and power spectrum. The final GNR-PS complex modified for tumor specificity serves as an excellent organ-specific theranostic probe for bioimaging and dual therapy both in vitro and in vivo. Principal component analysis designates photodynamic therapy a better candidate than that of photothermal therapy for long-term efficacy in vivo., (© 2019 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.)

In development, wound healing, and cancer metastasis, vertebrate cells move through 3D interstitial matrix, responding to chemical and physical guidance cues. Protrusion at the cell front has been extensively studied, but the retraction phase of the migration cycle is not well understood. Here, we show that fast-moving cells guided by matrix cues establish positive feedback control of rear retraction by sensing membrane tension. We reveal a mechanism of rear retraction in 3D matrix and durotaxis controlled by caveolae, which form in response to low membrane tension at the cell rear. Caveolae activate RhoA-ROCK1/PKN2 signaling via the RhoA guanidine nucleotide exchange factor (GEF) Ect2 to control local F-actin organization and contractility in this subcellular region and promote translocation of the cell rear. A positive feedback loop between cytoskeletal signaling and membrane tension leads to rapid retraction to complete the migration cycle in fast-moving cells, providing directional memory to drive persistent cell migration in complex matrices., (Copyright © 2019 The Authors. Published by Elsevier Inc. All rights reserved.)

The pattern of interactions between foot and mouth disease (FMD) viral protein 1 (VP1) with susceptible and resistant host integrins were deciphered. The putative effect of site-directed mutation on alteration of interaction is illustrated using predicted and validated 3D structures of VP1, mutated VP1 and integrins of Bos taurus , Gallus and Canis . Strong interactions were observed between FMDV-VP1 protein motifs at conserved tripeptide, Arg-Gly-Asp 143 RGD 145 and at domain 676 SIPLQ 680 in alpha-integrin of B. taurus. Notably, in-silico site-directed mutation in FMDV-VP1 protein led to complete loss of interaction between FMD-VP1 protein and B. taurus integrin, which confirmed the active role of arginine-glycine-aspartic acid (RGD) domain. Interestingly, in-vitro analysis demonstrates the persistence of the putative tropism site 'SIPLQ' in different cattle breeds undertaken. Thus, the attempt to decipher the tropism of FMDV at host receptor level interaction might be useful for future FMD control strategies through development of mimetic marker vaccines and/or host receptor manipulations. Communicated by Ramaswamy H. Sarma.

An aged circulatory environment can activate microglia, reduce neural precursor cell activity and impair cognition in mice. We hypothesized that brain endothelial cells (BECs) mediate at least some of these effects. We observe that BECs in the aged mouse hippocampus express an inflammatory transcriptional profile with focal upregulation of vascular cell adhesion molecule 1 (VCAM1), a protein that facilitates vascular-immune cell interactions. Concomitantly, levels of the shed, soluble form of VCAM1 are prominently increased in the plasma of aged humans and mice, and their plasma is sufficient to increase VCAM1 expression in cultured BECs and the hippocampi of young mice. Systemic administration of anti-VCAM1 antibody or genetic ablation of Vcam1 in BECs counteracts the detrimental effects of plasma from aged individuals on young brains and reverses aging aspects, including microglial reactivity and cognitive deficits, in the brains of aged mice. Together, these findings establish brain endothelial VCAM1 at the blood-brain barrier as a possible target to treat age-related neurodegeneration.

The spatial- and spectral-dependent optical reflectivity measurements are essential to characterize various natural as well as artificial micron-scale photonic nanostructures. However, it is onerous to measure spatially and spectrally resolved reflectivity values from such photonic nanostructures due to their size limitations. Here, we discuss the development of a versatile micro-reflectivity setup with an in situ optical microscope combined with high-resolution actuators to measure the reflectivity from areas as small as 25 × 25 µm 2 . We illustrate the reflectivity measurements from natural as well as artificially prepared ordered and disordered photonic nanostructures. The optical features that are hidden in the conventional reflectivity measurements are clearly resolved using the micro-reflectivity measurements. The proposed setup is also capable of measuring the polarization-dependent reflectivity and transmission of light.

Fundamental biological processes are carried out by curved epithelial sheets that enclose a pressurized lumen. How these sheets develop and withstand three-dimensional deformations has remained unclear. Here we combine measurements of epithelial tension and shape with theoretical modelling to show that epithelial sheets are active superelastic materials. We produce arrays of epithelial domes with controlled geometry. Quantification of luminal pressure and epithelial tension reveals a tensional plateau over several-fold areal strains. These extreme strains in the tissue are accommodated by highly heterogeneous strains at a cellular level, in seeming contradiction to the measured tensional uniformity. This phenomenon is reminiscent of superelasticity, a behaviour that is generally attributed to microscopic material instabilities in metal alloys. We show that in epithelial cells this instability is triggered by a stretch-induced dilution of the actin cortex, and is rescued by the intermediate filament network. Our study reveals a type of mechanical behaviour-which we term active superelasticity-that enables epithelial sheets to sustain extreme stretching under constant tension.

Recently, the nitrogen vacancy (NV) center has emerged as a deterministic single-photon source at room temperature. However, the presence of phonon sideband emission limits the use of NV centers in various quantum optical applications. Here we report a novel way to suppress the phonon sideband emission of NV centers aided with the enhancement of zero phonon line intensity using photonic crystals at room temperature. We study the photonic stop gap mediated charge state conversion in NV centers. The enhancement of phonon sideband emission from the same structures by implementing a different measurement geometry is also discussed. The studies open new avenues in studying the NV center-based quantum nanophotonics and biophotonics.

The integrin α 5 β 1 is overexpressed in colon, breast, ovarian, lung and brain tumours, and has been identified as key component in mechanosensing. In order to study how dynamic changes in α 5 β 1 engagement affect cellular behaviour, photoactivatable derivatives of α 5 β 1 -specific ligands are presented in this article. A photoremovable protecting group (PRPG) was introduced into the ligand structure at a relevant position for integrin recognition. The presence of the chromophore temporarily inhibited ligand bioactivity. Light exposure at a cell-compatible dose efficiently cleaved the protecting group and restored functionality. The photoactive ligand had an azide end-functional group for covalent immobilisation onto biomaterials by click chemistry. Selective cell responses (attachment, spreading, migration) to the activated ligand on the surface are achieved by controlled exposure to light, at similar levels to the native ligand. Spatial and temporal control of the cellular response is demonstrated, including the possibility of in situ activation. Photoactivatable integrin-selective ligands in model microenvironments will allow the study of cellular behaviour in response to changes in the activation of individual integrins as consequence of dynamic variations in matrix composition., (© 2018 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.)

Zinc, the essential trace element in human body exists either in the bound or free state, for both structural and functional roles. Insights on Zn 2+ distribution and its dynamics are essential in view of the fact that Zn 2+ dyshomeostasis is a risk factor for epileptic seizures, Alzheimer's disease, depression, etc. Herein, a bipyridine bridged bispyrrole (BP) probe is used for ratiometric imaging and quantification of Zn 2+ in hippocampal slices. The green fluorescence emission of BP shifts towards red in the presence of Zn 2+ . The probe is used to detect and quantify the exogenous and endogenous Zn 2+ in glioma cells and hippocampal slices. The dynamics of chelatable zinc ions during epileptic condition is studied in the hippocampal neurons, in vitro wherein the translocation of Zn 2+ from presynaptic to postsynaptic neuronal bodies is imaged and ratiometrically quantified. Raman mapping technique is used to confirm the dynamics of Zn 2+ under epileptic condition. Finally, the Zn 2+ distribution was imaged in vivo in epileptic rats and the total Zn 2+ in rat brain was quantified. The results favour the use of BP as an excellent Zn 2+ imaging probe in biological system to understand the zinc associated diseases and their management.

In the present investigation, the effect of timolol maleate loaded ocuserts was studied as an alternative for conventional anti-glaucoma formulation. Ocuserts were prepared using natural polymer sodium alginate and ethyl cellulose. Physico-chemical properties along with drug entrapment efficiency (94-98%), content uniformity (93.1% ± 0.264-98.00% ± 0.321), in vitro drug release (83.42% ± 0.35 at end of 12 h), ex vivo permeation all showed satisfactory results, which was found to follow zero order kinetics. Ex vivo permeation studies showed better results, revealed that the permeability coefficient was dependent on polymer type. The sterility test accelerated stability studies and in vivo studies such as eye irritancy test, in vivo drug release of the optimized ocusert was determined. The anti-glaucoma activity was measured using Schiotz tonometer at different time interval. Significant reduction in Intra ocular pressure (IOP) within 3 days was observed in case of rabbits treated with ocusert in comparison to the rabbit treated with marketed eye drop formulation. Hence timolol maleate loaded ocuserts proved to be a promising and viable alternative over conventional eye formulation for the sustained and controlled ophthalmic drug delivery, targeting the drug within the ocular globe thus improving patient compliance for the treatment of glaucoma., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Cancer, a condition with uncontrolled cell division, is the second leading cause of death worldwide. The currently available techniques for the imaging and treatment of cancer have their own limitations and hence a combination of more than one modality is expected to increase the efficacy of both diagnosis and treatment. In the present study, we have developed a multimodal imaging and therapeutic system by incorporating a chemotherapeutic drug, mitoxantrone (MTX) onto PEG coated gold nanorods (GNR). Strong absorption in the near-infrared (NIR) and visible regions qualifies GNR as an efficient photothermal (PTT) agent upon irradiation with either a NIR or visible laser. Additionally, the enhanced electric field of GNR makes it a suitable substrate for surface enhanced Raman scattering (SERS). Modification of GNR with amino PEG offers biocompatibility without affecting its optical property. In order to achieve tumor specificity, GNR-PEG was conjugated with tumor specific marker that can target cancer cells, leaving the normal cells unaffected. The incorporation of fluorescent chemotherapeutic drug mitoxantrone onto GNR-PEG facilitates chemotherapy as well as fluorescence imaging. The therapeutic efficacy of the developed GNR based system is tracked using fluorescence imaging and Raman imaging. The careful design of the system also facilitates the controlled release of the drug by photothermal triggering. Likewise, the imaging modality could be chosen as either Raman or fluorescence to monitor drug release in accordance with irradiation. The physico-chemical properties, and drug release profiles under different physiological conditions have been well studied. Finally, the developed system was tested for its therapeutic efficacy using cancer cells, in vitro. The receptor mediated cell uptake was more effective in folate receptor over-expressing cancer cells than in the normal and low-expressing cells. Accordingly the percentage of cell death was higher in folate receptor over-expressing cancer cells, which was further enhanced due to the effect of the dual therapeutic approach. The cell uptake and treatment efficacy was monitored using fluorescence microscopy and SERS. In conclusion, the developed GNR-PEG-MTX system is found to be an efficient multimodal therapeutic agent against cancer which could be tracked using two different techniques.

NF90 and splice variant NF110 are DNA- and RNA-binding proteins encoded by the Interleukin enhancer-binding factor 3 (ILF3) gene that have been established to regulate RNA splicing, stabilization and export. The roles of NF90 and NF110 in regulating transcription as chromatin-interacting proteins have not been comprehensively characterized. Here, chromatin immunoprecipitation followed by deep sequencing (ChIP-seq) identified 9,081 genomic sites specifically occupied by NF90/NF110 in K562 cells. One third of NF90/NF110 peaks occurred at promoters of annotated genes. NF90/NF110 occupancy colocalized with chromatin marks associated with active promoters and strong enhancers. Comparison with 150 ENCODE ChIP-seq experiments revealed that NF90/NF110 clustered with transcription factors exhibiting preference for promoters over enhancers (POLR2A, MYC, YY1). Differential gene expression analysis following shRNA knockdown of NF90/NF110 in K562 cells revealed that NF90/NF110 activates transcription factors that drive growth and proliferation (EGR1, MYC), while attenuating differentiation along the erythroid lineage (KLF1). NF90/NF110 associates with chromatin to hierarchically regulate transcription factors that promote proliferation and suppress differentiation.

Despite evidence that γδ T cells play an important role during malaria, their precise role remains unclear. During murine malaria induced by Plasmodium chabaudi infection and in human P. falciparum infection, we found that γδ T cells expanded rapidly after resolution of acute parasitemia, in contrast to αβ T cells that expanded at the acute stage and then declined. Single-cell sequencing showed that TRAV15N-1 (Vδ6.3) γδ T cells were clonally expanded in mice and had convergent complementarity-determining region 3 sequences. These γδ T cells expressed specific cytokines, M-CSF, CCL5, CCL3, which are known to act on myeloid cells, indicating that this γδ T cell subset might have distinct functions. Both γδ T cells and M-CSF were necessary for preventing parasitemic recurrence. These findings point to an M-CSF-producing γδ T cell subset that fulfills a specialized protective role in the later stage of malaria infection when αβ T cells have declined., (Copyright © 2018. Published by Elsevier Inc.)

In this Letter, we report on the design, fabrication, and implementation of a novel plasmon-mode-driven low-threshold near-infrared (NIR) random laser (RL) in the 850-900 nm range based on plasmonic ZnS@Au core-shell scatterers. Plasmon modes in the NIR region are used for nanoscale scatterer engineering of ZnS@Au core-shell particles to enhance scattering, as against pristine ZnS. This plasmonic scattering enhancement coupled with femtosecond (fs) laser pumping is shown to cause a three-fold lasing threshold reduction from 325  μJ/cm 2 to 100  μJ/cm 2 and a mode Q-factor enhancement from 200 to 540 for ZnS@Au-based RL, as compared to pristine ZnS-based RL. Local field enhancement due to plasmonic ZnS@Au scatterers, as evidenced in the finite-difference time-domain simulation, further adds to this enhancement. This work demonstrates a novel scheme of plasmonic mode coupling in the NIR region and fs excitation in a random laser photonic system, overcoming the inherent deficiencies of weak absorption of gain media and poor scattering cross sections of dielectric scatterers for random lasing in the NIR spectrum.

Graphene quantum dots (GQDs) are zero-dimensional materials that exhibit characteristics of both graphene and quantum dots. Herein, we report a rapid, relatively green, one-pot synthesis of size-tunable GQDs from graphene oxide (GO) by a sonochemical method with intermittent microwave heating, keeping the reaction temperature constant at 90 °C. The GQDs were synthesized by oxidative cutting of GO using KMnO 4 as an oxidizing agent within a short span of time (30 min) in an acid-free condition. The synthesized GQDs were of high quality and exhibited good quantum yield (23.8%), high product yield (>75%), and lower cytotoxicity (tested up to 1000 μg/mL). Furthermore, the as-synthesized GQDs were demonstrated as excellent fluorescent probes for bioimaging and label-free sensing of Fe(III) ions, with a detection limit as low as 10 × 10 -6 M., Competing Interests: The authors declare no competing financial interest.

Blood brain barrier (BBB) is a dynamic interface, comprising polarized endothelial cells, that separates the brain from the circulatory system. The highly protective nature of this tight junction impairs diagnosis and treatment of brain disorders. In this study, we designed a sub atomic size, near infrared emitting, dual function glutathione gold cluster with high fluorescence yield to facilitate permeability of BBB, for imaging applications and drug delivery. The gold cluster was then modified with Levodopa (l-dopa), to utilize the large amino acid transporter 1 (LAT1) pathways to enhance brain entry. Uptake and permeability of the nanoprobes were demonstrated using an established model of BBB, comprising brain endothelial cells (bEnd.3). The uptake and the clearance of l-dopa modified cluster was faster than the glutathione cluster. l-Dopa modified cluster supports the slow and sustained delivery of a model drug, pilocarpine, to the brain. Results of in vivo imaging and drug release in normal mice hold promise for considering the probe for early diagnosis of brain diseases, when the barrier is not disrupted, and for subsequent drug treatment.