Advancements in the development of fluorescent chemosensors based on CN bond isomerization/modulation mechanistic approaches.

The CN bond isomerization/modulation as a fluorescence signalling mechanism was explored by studying the photophysical properties of conformationally restricted molecules. From the beginning, the CN bond isomerization method has attracted the attention of researchers owing to its simplicity, high selectivity, and sensitivity in fluorescence evaluation. Continuous developments in the field of sensing using CN bond-containing compounds have been achieved via the customization of the isomerization process around the CN bond in numerous ways, and the results were obtained in the form of specific discrete photophysical changes. CN isomerization causes significant fluorescence enhancement in response to detected metal cations and other reactive species (Cys, Hys, ClO-, etc.) straightforwardly and effectively. This review sheds light on the process of CN bond isomerization/modulation as a signalling mechanism depending on fluorescence changes via conformational restriction. In addition, CN bond isomerization-based fluorescent sensors have yet to be well reviewed, although several fluorescent sensors based on this signalling mechanism have been reported. Therefore, CN-based fluorescent sensors are summarized in this review.

Sensory nerve and neuropeptide diversity in adipose tissues.

Both brown and white adipose tissues (BAT/WAT) are innervated by the peripheral nervous system, including efferent sympathetic nerves that communicate from the brain/central nervous system out to the tissue, and afferent sensory nerves that communicate from the tissue back to the brain and locally release neuropeptides to the tissue upon stimulation. This bidirectional neural communication is important for energy balance and metabolic control, as well as maintaining adipose tissue health through processes like browning (development of metabolically healthy brown adipocytes in WAT), thermogenesis, lipolysis, and adipogenesis. Decades of sensory nerve denervation studies have demonstrated the particular importance of adipose sensory nerves for brown adipose tissue and WAT functions, but far less is known about the tissue's sensory innervation compared to the better-studied sympathetic nerves and their neurotransmitter norepinephrine. In this review, we cover what is known and not yet known about sensory nerve activities in adipose, focusing on their effector neuropeptide actions in the tissue.

The metabolic and functional roles of sensory nerves in adipose tissues.

Homeostatic regulation of adipose tissue is critical for the maintenance of energy balance and whole-body metabolism. The peripheral nervous system provides bidirectional neural communication between the brain and adipose tissue, thereby providing homeostatic control. Most research on adipose innervation and nerve functions has been limited to the sympathetic nerves and their neurotransmitter norepinephrine. In recent years, more work has focused on adipose sensory nerves, but the contributions of subsets of sensory nerves to metabolism and the specific roles contributed by sensory neuropeptides are still understudied. Advances in imaging of adipose innervation and newer tissue denervation techniques have confirmed that sensory nerves contribute to the regulation of adipose functions, including lipolysis and browning. Here, we summarize the historical and latest findings on the regulation, function and plasticity of adipose tissue sensory nerves that contribute to metabolically important processes such as lipolysis, vascular control and sympathetic axis cross-talk.

Adenine nucleotide carrier protein dysfunction in human disease.

Adenine nucleotide translocase (ANT) is the prototypical member of the mitochondrial carrier protein family, primarily involved in ADP/ATP exchange across the inner mitochondrial membrane. Several carrier proteins evolutionarily related to ANT, including SLC25A24 and SLC25A25, are believed to promote the exchange of cytosolic ATP-Mg2+ with phosphate in the mitochondrial matrix. They allow a net accumulation of adenine nucleotides inside mitochondria, which is essential for mitochondrial biogenesis and cell growth. In the last two decades, mutations in the heart/muscle isoform 1 of ANT (ANT1) and the ATP-Mg2+ transporters have been found to cause a wide spectrum of human diseases by a recessive or dominant mechanism. Although loss-of-function recessive mutations cause a defect in oxidative phosphorylation and an increase in oxidative stress which drives the pathology, it is unclear how the dominant missense mutations in these proteins cause human diseases. In this review, we focus on how yeast was productively used as a model system for the understanding of these dominant diseases. We also describe the relationship between the structure and function of ANT and how this may relate to various pathologies. Particularly, mutations in Aac2, the yeast homolog of ANT, were recently found to clog the mitochondrial protein import pathway. This leads to mitochondrial precursor overaccumulation stress (mPOS), characterized by the toxic accumulation of unimported mitochondrial proteins in the cytosol. We anticipate that in coming years, yeast will continue to serve as a useful model system for the mechanistic understanding of mitochondrial protein import clogging and related pathologies in humans.

Pyrene appendant triazole-based chemosensors for sensing applications.

Over the last two decades, the design and development of fluorescent chemosensors for the targeted detection of heavy transition-metal (HTM) ions, anions, and biological analytes, have drawn much interest. Since the introduction of click chemistry in 2001, triazole moieties have become an increasingly prominent part of chemosensors. Triazoles generated via click reactions are crucial for sensing various ions and biological analytes. Recently, the number of studies in the field of pyrene appendant triazole moieties has risen dramatically, with more sophisticated and reliable triazole-containing chemosensors for various analytes of interest described. This review provides a general overview of pyrene appendant-triazole-based chemosensors that can detect a variety of metal cations, anions, and neutral analytes by using modular click-derived triazoles.

Age-related changes to adipose tissue and peripheral neuropathy in genetically diverse HET3 mice differ by sex and are not mitigated by rapamycin longevity treatment.

Neural communication between the brain and adipose tissues regulates energy expenditure and metabolism through modulation of adipose tissue functions. We have recently demonstrated that under pathophysiological conditions (obesity, diabetes, and aging), total subcutaneous white adipose tissue (scWAT) innervation is decreased ('adipose neuropathy'). With advanced age in the C57BL/6J mouse, small fiber peripheral nerve endings in adipose tissue die back, resulting in reduced contact with adipose-resident blood vessels and other cells. This vascular neuropathy and parenchymal neuropathy together likely pose a physiological challenge for tissue function. In the current work, we used the genetically diverse HET3 mouse model to investigate the incidence of peripheral neuropathy and adipose tissue dysregulation across several ages in both male and female mice. We also investigated the anti-aging treatment rapamycin, an mTOR inhibitor, as a means to prevent or reduce adipose neuropathy. We found that HET3 mice displayed a reduced neuropathy phenotype compared to inbred C56BL/6 J mice, indicating genetic contributions to this aging phenotype. Compared to female HET3 mice, male HET3 mice had worse neuropathic phenotypes by 62 weeks of age. Female HET3 mice appeared to have increased protection from neuropathy until advanced age (126 weeks), after reproductive senescence. We found that rapamycin overall had little impact on neuropathy measures, and actually worsened adipose tissue inflammation and fibrosis. Despite its success as a longevity treatment in mice, higher doses and longer delivery paradigms for rapamycin may lead to a disconnect between life span and beneficial health outcomes.

Synthesis, characterization, and electrocatalytic behaviour of hydrothermally grown nanostructured La2 O3 and La2 O3 /K-complex.

Rare earth metals play a conspicuous role in magnetic resonance imaging (MRI) for detecting cancerous cells. The alkali metal potassium is a neurotransmitter in the sodium-potassium pump in biomedical sciences. This unique property of rare earth metals and potassium drew our attention to carry forward this study. Therefore, in this work, previously synthesized potassium (K) complexes formed by the reflux of 4-N,N-dimethylaminobenzoic acid (DBA) and potassium hydroxide in methanol, and named [(µ2-4-N,N-dimethylaminobenzoate-κO)(µ2-4-N,N-dimethylaminobenzoic acid-κO)(4-N,N-dimethylaminobenzoic acid-κO) potassium(I) coordination polymer)] were treated hydrothermally with La2 O3 nanomaterials to obtain a nanohybrid La2 O3 /K-complex. After that, the K-complex was analyzed using single-crystal X-ray diffraction and 1 H and 13 C NMR spectroscopy. In addition, the structural and morphological properties of the as-prepared nanostructured La2 O3 /K-complex were also characterized, which involved an investigation using X-ray diffraction (XRD)spectroscopy, Fourier transform infrared (FTIR) spectroscopy, atomic force spectroscopy (AFM), transmission electron microscopy (TEM), and energy dispersive X-ray (EDX) analysis. After this, the electrochemical redox behaviour of the synthesized nanohybrid material was studied using cyclic voltammetry (CV) and differential pulse voltammetry (DPV). Therefore, the results from these studies revealed that the as-prepared material was a La2 O3 /K-complex that has a promising future role in sensing various analytes, as it showed effective electrocatalytic behaviour.

In vivo CRISPR screens reveal the landscape of immune evasion pathways across cancer.

The immune system can eliminate tumors, but checkpoints enable immune escape. Here, we identify immune evasion mechanisms using genome-scale in vivo CRISPR screens across cancer models treated with immune checkpoint blockade (ICB). We identify immune evasion genes and important immune inhibitory checkpoints conserved across cancers, including the non-classical major histocompatibility complex class I (MHC class I) molecule Qa-1b/HLA-E. Surprisingly, loss of tumor interferon-γ (IFNγ) signaling sensitizes many models to immunity. The immune inhibitory effects of tumor IFN sensing are mediated through two mechanisms. First, tumor upregulation of classical MHC class I inhibits natural killer cells. Second, IFN-induced expression of Qa-1b inhibits CD8+ T cells via the NKG2A/CD94 receptor, which is induced by ICB. Finally, we show that strong IFN signatures are associated with poor response to ICB in individuals with renal cell carcinoma or melanoma. This study reveals that IFN-mediated upregulation of classical and non-classical MHC class I inhibitory checkpoints can facilitate immune escape.

In vivo screens using a selective CRISPR antigen removal lentiviral vector system reveal immune dependencies in renal cell carcinoma.

CRISPR-Cas9 genome engineering has increased the pace of discovery for immunology and cancer biology, revealing potential therapeutic targets and providing insight into mechanisms underlying resistance to immunotherapy. However, endogenous immune recognition of Cas9 has limited the applicability of CRISPR technologies in vivo. Here, we characterized immune responses against Cas9 and other expressed CRISPR vector components that cause antigen-specific tumor rejection in several mouse cancer models. To avoid unwanted immune recognition, we designed a lentiviral vector system that allowed selective CRISPR antigen removal (SCAR) from tumor cells. The SCAR system reversed immune-mediated rejection of CRISPR-modified tumor cells in vivo and enabled high-throughput genetic screens in previously intractable models. A pooled in vivo screen using SCAR in a CRISPR-antigen-sensitive renal cell carcinoma revealed resistance pathways associated with autophagy and major histocompatibility complex class I (MHC class I) expression. Thus, SCAR presents a resource that enables CRISPR-based studies of tumor-immune interactions and prevents unwanted immune recognition of genetically engineered cells, with implications for clinical applications.

Microdissected "cuboids" for microfluidic drug testing of intact tissues.

As preclinical animal tests often do not accurately predict drug effects later observed in humans, most drugs under development fail to reach the market. Thus there is a critical need for functional drug testing platforms that use human, intact tissues to complement animal studies. To enable future multiplexed delivery of many drugs to one small biopsy, we have developed a multi-well microfluidic platform that selectively treats cuboidal-shaped microdissected tissues or "cuboids" with well-preserved tissue microenvironments. We create large numbers of uniformly-sized cuboids by semi-automated sectioning of tissue with a commercially available tissue chopper. Here we demonstrate the microdissection method on normal mouse liver, which we characterize with quantitative 3D imaging, and on human glioma xenograft tumors, which we evaluate after time in culture for viability and preservation of the microenvironment. The benefits of size uniformity include lower heterogeneity in future biological assays as well as facilitation of their physical manipulation by automation. Our prototype platform consists of a microfluidic circuit whose hydrodynamic traps immobilize the live cuboids in arrays at the bottom of a multi-well plate. Fluid dynamics simulations enabled the rapid evaluation of design alternatives and operational parameters. We demonstrate the proof-of-concept application of model soluble compounds such as dyes (CellTracker, Hoechst) and the cancer drug cisplatin. Upscaling of the microfluidic platform and microdissection method to larger arrays and numbers of cuboids could lead to direct testing of human tissues at high throughput, and thus could have a significant impact on drug discovery and personalized medicine.

Marginalizing health: employing an equity and intersectionality frame / Marginalizando a saúde: empregando uma abordagem de equidade e interseccionalidade

ABSTRACT The understanding that the drivers of inequities are multiple and intersecting is critical for health policy formulation and implementation. An intersectionality analysis reveals these relationships and allows a nuanced grasp of how health inequities are framed and understood. Using global statistics and other examples, the paper argues the significance of an intersectionality analysis in unravelling the disproportionate impact of inequity and the implications for the health and lives of persons experiencing these multiple discriminations. Attention to this, challenges the assumption of homogeneity and helps to visibilize lived realities. A few examples of acts of resistance are cited by the authors that have attempted to amplify the voices and knowledge of those whose realities are otherwise invisibilized by prevailing inequities, policies and discourses. 'Marginalizing' health thus implies an intersectional understanding of inequity as well as challenging and changing prevailing socio-political structures.

RESUMO O entendimento de que os fatores motivadores das desigualdades são múltiplos e se cruzam é fundamental para a formulação e para a implementação de políticas de saúde. Uma análise de interseccionalidade revela essas relações e permite uma compreensão diferenciada de como as iniquidades em saúde são estruturadas e compreendidas. Usando estatísticas globais e outros exemplos, o artigo argumenta a importância de uma análise de interseccionalidade para desvendar o impacto desproporcional da desigualdade e as implicações para a saúde e a vida das pessoas que sofrem essas múltiplas discriminações. Essa abordagem desafia o pressuposto de homogeneidade e ajuda a visibilizar as realidades vividas. Alguns exemplos de atos de resistência são citados pelos autores que tentaram ampliar as vozes e o conhecimento daqueles cujas realidades são, de outro modo, invisibilizadas pelas iniquidades, políticas e discursos predominantes. A 'marginalização' da saúde implica, portanto, um entendimento interseccional da desigualdade, assim como em enfrentar e mudar as estruturas sociopolíticas predominantes.

'Marginalizing' health: employing an equity and intersectionality frame / 'Marginalizando' a saúde: empregando uma abordagem de equidade e interseccionalidade

ABSTRACT The understanding that the drivers of inequities are multiple and intersecting is critical for health policy formulation and implementation. An intersectionality analysis reveals these relationships and allows a nuanced grasp of how health inequities are framed and understood. Using global statistics and other examples, the paper argues the significance of an intersectionality analysis in unravelling the disproportionate impact of inequity and the implications for the health and lives of persons experiencing these multiple discriminations. Attention to this, challenges the assumption of homogeneity and helps to visibilize lived realities. A few examples of acts of resistance are cited by the authors that have attempted to amplify the voices and knowledge of those whose realities are otherwise invisibilized by prevailing inequities, policies and discourses. 'Marginalizing' health thus implies an intersectional understanding of inequity as well as challenging and changing prevailing socio-political structures.

RESUMO O entendimento de que os fatores motivadores das desigualdades são múltiplos e se cruzam é fundamental para a formulação e para a implementação de políticas de saúde. Uma análise de interseccionalidade revela essas relações e permite uma compreensão diferenciada de como as iniquidades em saúde são estruturadas e compreendidas. Usando estatísticas globais e outros exemplos, o artigo argumenta a importância de uma análise de interseccionalidade para desvendar o impacto desproporcional da desigualdade e as implicações para a saúde e a vida das pessoas que sofrem essas múltiplas discriminações. Essa abordagem desafia o pressuposto de homogeneidade e ajuda a visibilizar as realidades vividas. Alguns exemplos de atos de resistência são citados pelos autores que tentaram ampliar as vozes e o conhecimento daqueles cujas realidades são, de outro modo, invisibilizadas pelas iniquidades, políticas e discursos predominantes. A 'marginalização' da saúde implica, portanto, um entendimento interseccional da desigualdade, assim como em enfrentar e mudar as estruturas sociopolíticas predominantes.

Fate of GdF3 nanoparticles-loaded PEGylated carbon capsules inside mice model: a step toward clinical application.

The successful translation of nanostructure-based bioimaging and/or drug delivery system needs extensive in vitro and in vivo studies on biocompatibility, biodistribution, clearance, and toxicity for its diagnostic applications. Herein, we have investigated the in vitro cyto-hemocompatibility, in vivo biodistribution, clearance, and toxicity in mice after systemic administration of GdF3 nanoparticles loaded PEGylated mesoporous carbon capsule (GdF3-PMCC)-based theranostic system. In vitro cyto-hemocompatibility study showed a very good biocompatibility up to concentration of 500 µg/ml. Biodistribution studies carried out from 1 h to 8 days showed that GdF3-PMCC was found in major organs, such as liver, kidney, spleen, and muscle till 4th day and it was negligible in any tissue after 8th day. The clearance study was carried out for a period of 8 days and it was observed that the urinary system is the main route of excretion of GdF3-PMCC. The tissue toxicity study was done for 15 days and histopathological analysis indicated that the GdF3-PMCC based theranostic system does not have any adverse effect in tissues. Thus, PMCCs are nontoxic and can be applied as theranostic agents in contrast to the other carbon-based systems (PEGylated carbon nanotubes and PEGylated graphene oxide) which showed significant toxicity.

Direct Intranuclear Anticancer Drug Delivery via Polydimethylsiloxane Nanoparticles: in Vitro and in Vivo Xenograft Studies.

Direct delivery of anticancer drugs to nuclei of tumor cells is required to enhance the therapeutic activity, which can be achieved by a nuclear localization signal (NLS) or peptide-decorated nanovehicles. However, NLS/peptide-based approaches may create certain undesirable immunological responses and the utilized synthesis processes are generally labor intensive. To this end, we report ligand-free, enhanced intranuclear delivery of Doxorubicin (Dox) to different cancer cells via porous polydimethylsiloxane (PDMS) nanoparticles (NPs). PDMS NPs were prepared by sacrificial silica template-based approach and Dox was loaded into the pores of PDMS NPs. These Dox-loaded PDMS NPs show enhanced cytotoxicity and reduce the IC50 values by 84 and 54% for HeLa and PC-3, respectively, compared to free Dox. Further, DNA damage in HeLa cells was estimated using comet assay suggesting enhanced DNA damage (72%) with Dox-loaded PDMS NPs as compared to free Dox (12%). The therapeutic efficiency of PDMS-Dox drug delivery system was tested in prostate cancer (PC-3) xenografts in NOD/SCID mice which showed enhanced tumor reduction (∼66%) as compared to free Dox. Taken together, our PDMS-Dox delivery system shows efficient and enhanced transportation of Dox to tumor cells which can be harnessed to develop advanced chemotherapy-based approaches to treat prostate and other cancers.

Multifunctional Mesoporous Carbon Capsules and their Robust Coatings for Encapsulation of Actives: Antimicrobial and Anti-bioadhesion Functions.

We present the synthesis and applications of multifunctional hollow porous carbon spheres with well-ordered pore architecture and ability to encapsulate functional nanoparticles. In the present work, the applications of hollow mesoporous carbon capsules (HMCCs) are illustrated in two different contexts. In the first approach, the hollow capsule core is used to encapsulate silver nanoparticles to impart antimicrobial characteristics. It is shown that silver-loaded HMCCs (concentration ∼100 µg/mL) inhibit the growth and multiplication of bacterial colonies of Escherichia coli (Gram-negative) and Staphylococcus aureus (Gram-positive) up to 96% and 83%, respectively. In the second part, the fabrication of hierarchical micro- and nanostructured superhydrophobic coatings of HMCCs (without encapsulation with silver nanoparticles) is evaluated for anti-bioadhesion properties. Studies of protein adsorption and microorganism and platelet adhesion have shown a significant reduction (up to 100%) for the HMCC-based superhydrophobic surfaces compared with the control surfaces. Therefore, this unique architecture of HMCCs and their coatings with the ability to encapsulate functional materials make them a promising candidate for a variety of applications.

PEGylated Carbon Nanocapsule: A Universal Reactor and Carrier for In Vivo Delivery of Hydrophobic and Hydrophilic Nanoparticles.

We have developed PEGylated mesoporous carbon nanocapsule as a universal nanoreactor and carrier for the delivery of highly crystalline hydrophobic/hydrophilic nanoparticles (NPs) which shows superior biocompatibility, dispersion in body fluids, good biodistribution and NPs independent cellular uptake mechanism. The hydrophobic/hydrophilic NPs without surface modification were synthesized in situ inside the cavities of mesoporous carbon capsules (200-850 nm). Stable and inert nature of carbon capsules in a wide range of reaction conditions like high temperature and harsh solvents, make it suitable for being used as nano/microreactors for the syntheses of a variety of NPs for bioimaging applications, such as NaYF4:Eu(3+)(5%), LaVO4:Eu(3+)(10%), GdVO4:Eu(3+)(10%), Y2O3:Eu(3+)(5%), GdF3:Tb(3+)(10%), Mo, Pt, Pd, Au, and Ag. Multiple types of NPs (Y2O3:Eu(3+)(5%) (hydrophobic) and GdF3:Tb(3+)(10%) (hydrophilic)) were coloaded inside the carbon capsules to create a multimodal agent for magneto-fluorescence imaging. Our in vivo study clearly suggests that carbon capsules have biodistribution in many organs including liver, heart, spleen, lungs, blood pool, and muscles.

A 2-(2'-hydroxyphenyl)benzothiazole (HBT)-quinoline conjugate: a highly specific fluorescent probe for Hg(2+) based on ESIPT and its application in bioimaging.

A benzothiazole derived chemosensor L has been designed based on the excited-state intramolecular proton transfer (ESIPT) mechanism to afford a fluorescence turn-on response specifically in the presence of Hg(2+) ions over a host of biologically relevant metal ions as well as toxic heavy metal ions. The chemosensor exhibits high sensitivity with the detection limit down to 0.11 µM. The metal binding is supported by (1)H NMR titrations, ESI-MS spectral analysis and substantiated by theoretical calculations using the density functional theory. The probe shows cell membrane permeability and efficiency for the detection of Hg(2+) in HeLa cells.