scRNA-seq profiling of human granulocytes reveals expansion of developmentally flexible neutrophil precursors with mixed neutrophil and eosinophil properties in asthma.

Neutrophils and eosinophils share common hematopoietic precursors and usually diverge into distinct lineages with unique markers before being released from their hematopoietic site, which is the bone marrow (BM). However, previous studies identified an immature Ly6g(+) Il-5Rα(+) neutrophil population in mouse BM, expressing both neutrophil and eosinophil markers suggesting hematopoietic flexibility. Moreover, others have reported neutrophil populations expressing eosinophil-specific cell surface markers in tissues and altered disease states, confusing the field regarding eosinophil origins, function, and classification. Despite these reports, it is still unclear whether hematopoietic flexibility exists in human granulocytes. To answer this, we utilized single-cell RNA sequencing (scRNA-seq) and CITE-seq to profile human BM and circulating neutrophils and eosinophils at different stages of differentiation and determine whether neutrophil plasticity plays role in asthmatic inflammation. We show that immature metamyelocyte neutrophils in humans expand during severe asthmatic inflammation and express both neutrophil and eosinophil markers. We also show an increase in tri-lobed eosinophils with mixed neutrophil and eosinophil markers in allergic asthma and that IL-5 promotes differentiation of immature blood neutrophils into tri-lobed eosinophilic phenotypes suggesting a mechanism of emergency granulopoiesis to promote myeloid inflammatory or remodeling response in patients with chronic asthma. By providing insights into unexpectedly flexible granulocyte biology and demonstrating emergency hematopoiesis in asthma, our results highlight the importance of granulocyte plasticity in eosinophil development and allergic diseases.

Wearable and Noninvasive Device for Integral Congestive Heart Failure Management in the IoMT Paradigm.

Noninvasive remote monitoring of hemodynamic variables is essential in optimizing treatment opportunities and predicting rehospitalization in patients with congestive heart failure. The objective of this study is to develop a wearable bioimpedance-based device, which can provide continuous measurement of cardiac output and stroke volume, as well as other physiological parameters for a greater prognosis and prevention of congestive heart failure. The bioimpedance system, which is based on a robust and cost-effective measuring principle, was implemented in a CMOS application specific integrated circuit, and operates as the analog front-end of the device, which has been provided with a radio-frequency section for wireless communication. The operating parameters of the proposed wearable device are remotely configured through a graphical user interface to measure the magnitude and the phase of complex impedances over a bandwidth of 1 kHz to 1 MHz. As a result of this study, a cardiac activity monitor was implemented, and its accuracy was evaluated in 33 patients with different heart diseases, ages, and genders. The proposed device was compared with a well-established technique such as Doppler echocardiography, and the results showed that the two instruments are clinically equivalent.

Molecular basis of differential HLA class I-restricted T cell recognition of a highly networked HIV peptide.

Cytotoxic-T-lymphocyte (CTL) mediated control of HIV-1 is enhanced by targeting highly networked epitopes in complex with human-leukocyte-antigen-class-I (HLA-I). However, the extent to which the presenting HLA allele contributes to this process is unknown. Here we examine the CTL response to QW9, a highly networked epitope presented by the disease-protective HLA-B57 and disease-neutral HLA-B53. Despite robust targeting of QW9 in persons expressing either allele, T cell receptor (TCR) cross-recognition of the naturally occurring variant QW9_S3T is consistently reduced when presented by HLA-B53 but not by HLA-B57. Crystal structures show substantial conformational changes from QW9-HLA to QW9_S3T-HLA by both alleles. The TCR-QW9-B53 ternary complex structure manifests how the QW9-B53 can elicit effective CTLs and suggests sterically hindered cross-recognition by QW9_S3T-B53. We observe populations of cross-reactive TCRs for B57, but not B53 and also find greater peptide-HLA stability for B57 in comparison to B53. These data demonstrate differential impacts of HLAs on TCR cross-recognition and antigen presentation of a naturally arising variant, with important implications for vaccine design.

The role of titanium-oxo clusters in the sulfate process for TiO2 production.

TiO2 is manufactured for white pigments, solar cells, self-cleaning surfaces and devices, and other photocatalytic applications. The industrial synthesis of TiO2 entails: (1) the dissolution of ilmenite ore (FeTiO3) in aqueous sulfuric acid which precipitates the Fe while retaining the Ti in solution, followed by (2) dilution or heating the Ti sulfate solution to precipitate the pure form of TiO2. The underlying chemistry of these processing steps remains poorly understood. Here we show that the dissolution of a simple TiIV-sulfate salt, representative of the industrial sulfate process for the production of TiO2, immediately self-assembles into a soluble Ti-octadecameric cluster, denoted as {Ti18}. We observed {Ti18} in solution by small-angle X-ray scattering and Ti extended X-ray absorption fine structure (Ti-EXAFS) analysis, and ultimately crystallized it for absolute identification. The {Ti18} metal-oxo cluster was previously reported as a polycation; but shown here, it can also be a polyanion, dependent on the number of sulfate ligands it carries. After immediate self-assembly, the {Ti18}-cluster persists until TiO2 precipitates, with no easily identified structural intermediates in the solution or solid state, despite the fact that the atomic arrangement of {Ti18} differs vastly from that of titania. The evolution from solution phase {Ti18} to precipitated TiO2 nanoparticles was detailed by X-ray scattering and Ti-EXAFS. We offer a hypothesis for the key mechanism of complete separation of Fe from Ti in the industrial sulfate process. These findings also highlight the emerging importance of the unusual Ti(Ti)5 pentagonal building unit, featured in {Ti18} as well as other early d0 transition metal-oxo clusters including Nb, Mo and W. Finally, this study presents an example of crystal growth mechanisms in which the observed "pre-nucleation cluster" does not necessarily predicate the structure of the precipitated solid.

HIV Controllers Exhibit Effective CD8+ T Cell Recognition of HIV-1-Infected Non-activated CD4+ T Cells.

Even with sustained antiretroviral therapy, resting CD4+ T cells remain a persistent reservoir of HIV infection, representing a critical barrier to curing HIV. Here, we demonstrate that CD8+ T cells recognize infected, non-activated CD4+ T cells in the absence of de novo protein production, as measured by immune synapse formation, degranulation, cytokine production, and killing of infected cells. Immune recognition is induced by HLA-I presentation of peptides derived from incoming viral particles, and recognition occurred either following cell-free virus infection or following cell-to-cell spread. CD8+ T cells from HIV controllers mediate more effective immune recognition than CD8+ T cells from progressors. These results indicate that non-activated HIV-infected CD4+ T cells can be targeted by CD8+ T cells directly after HIV entry, before reverse transcription, and thus before the establishment of latency, and suggest a mechanism whereby the immune response may reduce the size of the HIV reservoir.

Enhancement of anion recognition exhibited by a zinc-imidazole-based ion-pair receptor composed of C-H hydrogen- and halogen-bond donor groups.

A 2-haloimidazole-tetraphenylethylene ion-pair receptor 1 is shown to recognise only HSO4- anions in the presence of a cobound Zn2+ cation guest species, which induced a remarkable increase with concomitant blue shift of the emission band of the complex [1·2Zn]4+ whereas no affinity of the free receptor 1 by the anions is observed. In addition, the downfield shifts observed by 1H NMR of the Ha, Hb and Hc protons of the complex [1·2Zn]4+ upon the addition of HSO4- anions indicate their participation in the recognition event. According to DFT studies, upon chelating a Zn2+ cation with two imidazole nitrogen atoms, receptor 1 adopts a conformation ideally fitted to recognise HSO4- through a combination of C(sp2)-HO and C(sp3)-HO hydrogen bondings, C+(sp2)-BrO halogen bonding and C(sp2)O tetrel bonding.

T cell receptors for the HIV KK10 epitope from patients with differential immunologic control are functionally indistinguishable.

HIV controllers (HCs) are individuals who can naturally control HIV infection, partially due to potent HIV-specific CD8+ T cell responses. Here, we examined the hypothesis that superior function of CD8+ T cells from HCs is encoded by their T cell receptors (TCRs). We compared the functional properties of immunodominant HIV-specific TCRs obtained from HLA-B*2705 HCs and chronic progressors (CPs) following expression in primary T cells. T cells transduced with TCRs from HCs and CPs showed equivalent induction of epitope-specific cytotoxicity, cytokine secretion, and antigen-binding properties. Transduced T cells comparably, albeit modestly, also suppressed HIV infection in vitro and in humanized mice. We also performed extensive molecular dynamics simulations that provided a structural basis for similarities in cytotoxicity and epitope cross-reactivity. These results demonstrate that the differential abilities of HIV-specific CD8+ T cells from HCs and CPs are not genetically encoded in the TCRs alone and must depend on additional factors.

Interlocked Supramolecular Polymers Created by Combination of Halogen- and Hydrogen-Bonding Interactions through Anion-Template Self-Assembly.

We present the synthesis and oxoanion-assembling properties of a monomer with a naphthalene ring as a central core decorated with two arms containing iodotriazolium rings as anion binding sites. Interactions with SO42-, H2PO4-, and HP2O73- anions, via a cooperative mechanism, afforded new supramolecular materials stabilized by a combination of halogen- and hydrogen-bonding interactions. 1H NMR experiments and solid-state structure provided evidence for the initial formation of a supramolecular linear chain, nucleation step, and then two different supramolecular chains are interpenetrated with each other, elongation steps, involving the formation of hydrogen bonds between two oxygens of the anion from one of the chains and the naphthalene inner protons from the other chain. Scanning electron microscopy studies revealed that the morphology of the crystals changed dramatically with the nature of the anion added.

Modulation of the Selectivity in Anions Recognition Processes by Combining Hydrogen- and Halogen-Bonding Interactions.

Most of the halogen bonding receptors for anions described use halogen bonding binding sites solely in the anion recognition process; only a few examples report the study of anion receptors in which the halogen bonding interaction has been used in combination with any other non-covalent interaction. With the aims to extend the knowledge in the behaviour of this kind of mixed receptors, we report here the synthesis and the anion recognition and sensing properties of a new halogen- and hydrogen- bonding receptor which binds anions by the cooperation of both non-covalent interactions. Fluorescence studies showed that the behaviour observed in the anion recognition sensing is similar to the one previously described for the halogen analogue and is quite different to the hydrogen one. On the other hand, the association constants obtained by ¹H-NMR data demonstrate that the mixed halogen- and hydrogen-bonding receptor is more selective for SO42- anion than the halogen or hydrogen analogues.

Aqueous Bismuth Titanium-Oxo Sulfate Cluster Speciation and Crystallization.

Inorganic aqueous metal-oxo clusters are both functional "molecular metal oxides" and intermediates to understand metal oxide growth from water. There has been a recent surge in discovery of aqueous Ti-oxo clusters but without extensive solution characterization. We use small-angle and total X-ray scattering, dynamic light scattering, transmission electron microscopy, and a single-crystal X-ray structure to show that heterometals such as bismuth stabilize labile Ti-oxo sulfate clusters in aqueous solution.[Ti22 Bi7 O41 (OH)(OH2 )30 (SO4 )12 ]2+ features edge-sharing between the Ti and Bi polyhedra, in contrast to the dominant corner-linking of Ti-oxo clusters. Bi stabilizes the Ti-polyhedra, which are synergistically stabilized by the bidentate sulfates. Gained stability and potential functionality from heterometals is an incentive to develop more broadly the landscape of heterometallic Ti-oxo clusters.

Anion Recognition Strategies Based on Combined Noncovalent Interactions.

This review highlights the most significant examples of an emerging field in the design of highly selective anion receptors. To date, there has been remarkable progress in the binding and sensing of anions. This has been driven in part by the discovery of ways to construct effective anion binding receptors using the dominant N-H functional groups and neutral and cationic C-H hydrogen bond donors, as well as underexplored strong directional noncovalent interactions such as halogen-bonding and anion-π interactions. In this review, we will describe a new and promising strategy for constructing anion binding receptors with distinct advantages arising from their elaborate design, incorporating multiple binding sites able to interact cooperatively with anions through these different kinds of noncovalent interactions. Comparisons with control species or solely hydrogen-bonding analogues reveal unique characteristics in terms of strength, selectivity, and interaction geometry, representing important advances in the rising field of supramolecular chemistry.

Encapsulation of a {Cu16} cluster containing four [Cu4O4] cubanes within an isopolyoxometalate {W44} cluster.

We report a {Cu16} embedded within a {W44} cluster containing four cubane-like [Cu4O4] units within an isopolyoxotungstate (isoPOT) in a {Na4Cu4[(H2W11O38) (CH3COO)(OH)3]}4·88H2O (1) and a polyanion Cu-linked {W11} chain Na6Cu2[(H2W11O38)(CH3COO)(OH)]·26H2O (2). Electronically, the redox properties show that both compounds 1 and 2 undergo irreversible reductions resulting in the demetalation of the compounds, whilst the magnetic behavior of 1 and 2 shows a weak antiferromagnetic and a stronger ferromagnetic coupling, respectively.

Electronic and relativistic contributions to ion-pairing in polyoxometalate model systems.

Ion pairs and solubility related to ion-pairing in water influence many processes in nature and in synthesis including efficient drug delivery, contaminant transport in the environment, and self-assembly of materials in water. Ion pairs are difficult to observe spectroscopically because they generally do not persist unless extreme solution conditions are applied. Here we demonstrate two advanced techniques coupled with computational studies that quantify the persistence of ion pairs in simple solutions and offer explanations for observed solubility trends. The system of study, ([(CH3)4N]+,Cs)8[M6O19] (M = Nb,Ta), is a set of unique polyoxometalate salts whose water solubility increases with increasing ion-pairing, contrary to most ionic salts. The techniques employed to characterize Cs+ association with [M6O19]8- and related clusters in simple aqueous media are 133Cs NMR (nuclear magnetic resonance) quadrupolar relaxation rate and PDF (pair distribution function) from X-ray scattering. The NMR measurements consistently showed more extensive ion-pairing of Cs+ with the Ta-analogue than the Nb-analogue, although the electrostatics of the ions should be identical. Computational studies also ascertained more persistent Cs+-[Ta6O19] ion pairs than Cs+-[Nb6O19] ion pairs, and bond energy decomposition analyses determined relativistic effects to be the differentiating factor between the two. These distinctions are likely responsible for many of the unexplained differences between aqueous Nb and Ta chemistry, while they are so similar in the solid state. The X-ray scattering studies show atomic level detail of this ion association that has not been prior observed, enabling confidence in our structures for calculations of Cs-cluster association energies. Moreover, detailed NMR studies allow quantification of the number of Cs+ associated with a single [Nb6O19]8- or [Ta6O19]8- anion which agrees with the PDF analyses.

A genome-wide CRISPR screen identifies a restricted set of HIV host dependency factors.

Host proteins are essential for HIV entry and replication and can be important nonviral therapeutic targets. Large-scale RNA interference (RNAi)-based screens have identified nearly a thousand candidate host factors, but there is little agreement among studies and few factors have been validated. Here we demonstrate that a genome-wide CRISPR-based screen identifies host factors in a physiologically relevant cell system. We identify five factors, including the HIV co-receptors CD4 and CCR5, that are required for HIV infection yet are dispensable for cellular proliferation and viability. Tyrosylprotein sulfotransferase 2 (TPST2) and solute carrier family 35 member B2 (SLC35B2) function in a common pathway to sulfate CCR5 on extracellular tyrosine residues, facilitating CCR5 recognition by the HIV envelope. Activated leukocyte cell adhesion molecule (ALCAM) mediates cell aggregation, which is required for cell-to-cell HIV transmission. We validated these pathways in primary human CD4+ T cells through Cas9-mediated knockout and antibody blockade. Our findings indicate that HIV infection and replication rely on a limited set of host-dispensable genes and suggest that these pathways can be studied for therapeutic intervention.

Preparation and sensing properties of a nitrogen-rich ferrocene-imidazole-quinoxaline triad decorated with pyrrole rings.

The synthesis and sensing properties of the nitrogen-rich ferrocene-imidazole-quinoxaline triad 1 decorated with two pyrrole rings have been described. Due to its ditopic nature, this molecule behaves as an ion-pair receptor for Ni2+cations and AcO- anions, although no affinity for either of the discrete ions is observed. It also displays the rare property consistent with the cooperative AND recognition of ion pairs. Thus, this receptor shows an important enhancement for binding AcO- anions when it is co-bound to Hg2+cations, whereas no affinity for the free receptor by the anion is observed.

Chemical Stabilization and Electrochemical Destabilization of the Iron Keggin Ion in Water.

The iron Keggin ion is identified as a structural building block in both magnetite and ferrihydrite, two important iron oxide phases in nature and in technology. Discrete molecular forms of the iron Keggin ion that can be both manipulated in water and chemically converted to the related metal oxides are important for understanding growth mechanisms, in particular, nonclassical nucleation in which cluster building units are preserved in the aggregation and condensation processes. Here we describe two iron Keggin ion structures, formulated as [Bi6FeO4Fe12O12(OH)12(CF3COO)10(H2O)2]3+ (Kegg-1) and [Bi6FeO4Fe12O12(OH)12(CF3COO)12]1+ (Kegg-2). Experimental and simulated X-ray scattering studies show indefinite stability of these clusters in water from pH 1-3. The tridecameric iron Keggin-ion core is protected from hydrolysis by a synergistic effect of the capping Bi3+ cations and the trifluoroacetate ligands that, respectively, bond to the iron and bridge to the bismuth. By introducing electrons to the aqueous solution of clusters, we achieve complete separation of bismuth from the cluster, and the iron Keggin ion rapidly converts to magnetite and/or ferrihydrite, depending on the mechanism of reduction. In this strategy, we take advantage of the easily accessible reduction potential and crystallization energy of bismuth. Reduction was executed in bulk by chemical means, by voltammetry, and by secondary effects of transmission electron microscopy imaging of solutions. Prior, we showed a less stable analogue of the iron Keggin cluster converted to ferrihydrite simply upon dissolution. The prior and currently studied clusters with a range of reactivity provide a chemical system to study molecular cluster to metal oxide conversion processes in detail.

Comparative Study of Charge-Assisted Hydrogen- and Halogen-Bonding Capabilities in Solution of Two-Armed Imidazolium Receptors toward Oxoanions.

Two-armed imidazolium-based anion receptors have been prepared. The central 2,7-disubstituted naphthalene ring features two photoactive anthracene end-capped side arms with central 2-bromoimidazolium or hydrogen-bonding imidazolium receptors. Combined emission and (1)H and (31)P NMR studies carried out in the presence of a wide variety of anions reveal that only HP2O7(3-), H2PO4(-), SO4(2-), and F(-) anions promoted noticeable changes. The halogen receptor 6(2+)·2PF6(-) acts as a selective fluorescent molecular sensor for H2PO4(-) anions, since only this anion promotes the appearance of the anthracene excimer emission band, whereas it remains unchanged in the presence of the other tested anions. In addition this halogen receptor behaves as a chemodosimeter toward HP2O7(3-) anion, through its transformation into the corresponding bis-imidazolone after debromination by the action of the basic anion. The association constant values of the halogen-bonding complexes in a competitive solvent CD3CN/MeOD (8/2) mixture with H2PO4(-) and SO4(2-) anions are higher than those found for the hydrogen-bonding counterpart. In contrast, in the less competitive CH3CN solvent higher binding affinity for anions corresponds to the hydrogen-bonding receptor 7(2+)·2PF6(-). In addition, the receptor 6(2+)·2PF6(-) represents a useful alternative as an imaging agent in living cells in a wide range of emission wavelengths.

Open conformers of HLA-F are high-affinity ligands of the activating NK-cell receptor KIR3DS1.

The activating natural killer (NK)-cell receptor KIR3DS1 has been linked to the outcome of various human diseases, including delayed progression of disease caused by human immunodeficiency virus type 1 (HIV-1), yet a ligand that would account for its biological effects has remained unknown. We screened 100 HLA class I proteins and found that KIR3DS1 bound to HLA-F, a result we confirmed biochemically and functionally. Primary human KIR3DS1(+) NK cells degranulated and produced antiviral cytokines after encountering HLA-F and inhibited HIV-1 replication in vitro. Activation of CD4(+) T cells triggered the transcription and surface expression of HLA-F mRNA and HLA-F protein, respectively, and induced binding of KIR3DS1. HIV-1 infection further increased the transcription of HLA-F mRNA but decreased the binding of KIR3DS1, indicative of a mechanism for evading recognition by KIR3DS1(+) NK cells. Thus, we have established HLA-F as a ligand of KIR3DS1 and have demonstrated cell-context-dependent expression of HLA-F that might explain the widespread influence of KIR3DS1 in human disease.

[End stage renal disease lymphopenia; characterization and clinical correlation]. / Caracterización y correlación clínica de la linfopenia en la enfermedad renal en estadio terminal.

BACKGROUND: Patients with End Stage Renal Disease (ESRD), defined as those in Stage 5 of the Kidney Disease Outcome Quality Initiative (KDOQI) classification, have a number of acquired immune deficiencies secondary to the uremic stage, among them lymphopenia. In the present report, we retrospectively characterized the peripheral blood lymphocyte counts in a group of patients with ESRD and we related lymphopenia to their biochemical parameters and to the presence or absence of infections. METHODS: Medical records from 190 patients in ESRD were selected from 282 medical records of patients being treated between February 2008 and November 2012 for kidney failure at different stages. A number of variables, including lymphocyte counts, biochemical parameters and infections, were analyzed at two different time points: Before and during dialysis. RESULTS: ESRD patients analyzed had a well defined peripheral blood hematological pattern, characterized by severe chronic anemia, normal or elevated leukocyte count and normal or below normal lymphocyte count. The degree of hematological changes correlated with the depth of renal dysfunction and improved with dialysis along with the improvement of urea and creatinine values. CONCLUSIONS: Lymphopenia was present in around half ESRD patients and was associated with increased infections, but they were of the same type as those present in ESRD patients without lymphopenia. Infections were different as those commonly associated with other immune deficiency lymphopenias. The implications of these findings are discussed.

Introducción: los pacientes con insuficiencia renal crónica en etapa terminal (ERET), presentan alteraciones inmunológicas diversas que los hacen más susceptibles a infecciones. Entre las alteraciones reportadas se encuentra la linfopenia. Se han realizado pocos estudios en nuestro medio que muestren la frecuencia y características de esta alteración así como su trascendencia clínica, relacionada con las infecciones que afectan a estos pacientes. Métodos: se analizó una serie de variables, incluyendo los valores de linfocitos y la presencia de infecciones, en un grupo de 190 pacientes con ERET de febrero 2008 a noviembre 2012. Se correlacionan y comparan los valores obtenidos entre ellos en dos momentos de su evolución: antes y durante su tratamiento dialítico. Resultados: en los pacientes con ERET, se obtuvo un perfil hematológico característico, caracterizado por anemia crónica severa, leucocitos totales normales o por arriba de lo normal y linfocitos normales o por debajo de lo normal (linfopenia). El grado de alteración hematológica correlacionó con el grado de afección renal y se corrigió en la medida que se corrigieron las alteraciones bioquímicas relacionadas con la ERET mediante diálisis peritoneal. Conclusiones: la linfopenia se encontró en cerca de la mitad de los pacientes con ERET y se asoció con el incremento de infecciones; el tipo de infecciones fue similar a lo observado en pacientes sin linfopenia y diferente al observado en pacientes con inmunodeficiencias primarias o adquiridas que afectan a los linfocitos.

Host-Guest Chemistry: Oxoanion Recognition Based on Combined Charge-Assisted C-H or Halogen-Bonding Interactions and Anion⋠⋠⋠Anion Interactions Mediated by Hydrogen Bonds.

Several bis-triazolium-based receptors have been synthesized and their anion-recognition capabilities have been studied. The central chiral 1,1'-bi-2-naphthol (BINOL) core features either two aryl or ferrocenyl end-capped side arms with central halogen- or hydrogen-bonding triazolium receptors. NMR spectroscopic data indicate the simultaneous occurrence of several charge-assisted aliphatic and heteroaromatic C-H noncovalent interactions and combinations of C-H hydrogen and halogen bonding. The receptors are able to selectively interact with HP2 O7 (3-) , H2 PO4 (-) , and SO4 (2-) anions, and the value of the association constant follows the sequence: HP2 O7 (3-) >SO4 (2-) >H2 PO4 (-) . The ferrocenyl end-capped 7(2+) ⋅2 BF4 (-) receptor allows recognition and differentiation of H2 PO4 (-) and HP2 O7 (3-) anions by using different channels: H2 PO4 (-) is selectively detected through absorption and emission methods and HP2 O7 (3-) by using electrochemical techniques. Significant structural results are the observation of an anion⋅⋅⋅anion interaction in the solid state (2:2 complex, 6(2+) ⋅[H2 P2 O7 ](2-) ), and a short C-I⋅⋅⋅O contact is observed in the structure of the complex [8(2+) ][SO4 ]0.5 [BF4 ].