Noninvasive Detection of Bladder Cancer Markers Based on Gold Nanomushrooms and Sandwich Immunoassays

Bladder cancer is one of the most common malignancies in the urinary system. Cystoscopy is the traditional standard diagnostic method for bladder cancer with subsequent biopsy or surgery. However, this method is uncomfortable for most patients because it requires anesthesia and possibly causes infections. Because of the high recurrence rate of bladder cancer, a rapid, low-cost, high-sensitivity, and noninvasive sensing method is needed. This study employed gold nanomushroom (AuNM) chips for bladder cancer biomarker detection, combining the benefits of sandwich immunoassay and localized surface plasmon resonance (LSPR) sensing. With a metal nanotransfer printing technique, which is cheap and straightforward, the AuNMs were patterned on flexible polycarbonate (PC) sheets. The gold caps stood above PC stems and provided ample spatial areas for capturing the biomarkers to be sensed. Three biomarkers served as the antigens and analytes, including human complement factor H (CFH), hyaluronic acid (HA), and nuclear matrix protein 22 (NMP22). Different antibodies, against the same biomarker, were covalently conjugated to AuNMs or gold nanoparticles, respectively. When the antibody–antigen–antibody sandwich structure formed, the plasmonic coupling between the AuNM surface and the gold nanoparticles significantly enhanced LSPR signals. The LSPR red shifts correlated quantitatively with the concentrations of the biomarkers. The limits of detection were 6.5, 8.3, and 7.0 pg/mL for CFH, HA, and NMP22, respectively. The chip’s specificity was tested and confirmed, excluding the nonspecific binding and false-positive possibility. The sensing performance of this sandwich immunoassay-based AuNM chip was better than that of the commercialized enzyme-linked immunosorbent assay. It provided a rapid, label-free, and easy operating platform for diagnosing and monitoring bladder cancer

Stereoselective Glycosylation of <i>exo</i>-Glycals Accelerated by Ferrier-Type Rearrangement

Owing to the driving force of the Ferrier-type rearrangement, the exo-glycals are highly reactive with various alcohols to afford glycosides and glycoconjugates with exclusive α-configuration. The resulting vinyl group in these glycosylation products can be further elaborated for general applications, including the synthesis of spiro derivatives and glycosylation of 2-ketoaldonic acids

Regeneration of PAPS for the Enzymatic Synthesis of Sulfated Oligosaccharides

This paper describes the study of 3‘-phosphoadenosine-5‘-phosphosulfate (PAPS) regeneration from 3‘-phosphoadenosine-5‘-phosphate (PAP) for use in practical syntheses of carbohydrate sulfates which are catalyzed by sulfotransferases. Among the regeneration systems, the one with recombinant aryl sulfotransferase proved to be the most practical. This regeneration system was coupled with a sulfotransferase-catalyzed reaction, using a recombinant Nod factor sulfotransferase, for the synthesis of various oligosaccharide sulfates that were further glycosylated using glycosyltransferases

Conservation of the L4 regions among mammalian galectins.

<p>Structure-based sequence alignment of S4-S6 β-strands of mammalian galectin-1s (A), galectin-3s (B), galectin-7s (C) and galectin-2s (D). According to the variation at corresponding position of Glu58^hGal7, mammalian galectin-7s are further divided into two subgroups, the hGal7 group and hGal7-llike group. Sequence comparison of L4 regions of human Galectin-1, 3 and mammalian Galectin-2, 7 (E). Invariant LNs-contacting residues of galectins are indicated by asterisks. Residues involved in unique salt-bridge network are colored according to negative (red) or positive (blue) charged properties. The sequences of mammalian galectins were selected from human (<i>Homo sapiens</i>: galectin-1 [NP_002296.1], galectin-2 [NP_006489.1], galectin-3 [BAA22164.1] and galectin-7 [NP_002298.1]), Gorilla (<i>Gorilla gorilla gorilla</i>: galectin-1 [XP_004063482.1], galectin-2 [XP_004063485.1], galectin-3 [XP_004055252.1] and galectin-7 [XP_004060726.1]), Monkey (<i>Macaca mulatta</i>: galectin-1 [NP_001162098.1], galectin-2 [XP_001087063.1], galectin-3 [NP_001253292.1] and galectin-7 [NP_001083444.1]), Rat (<i>Rattus norvegicus</i>: galectin-1 [NP_063969.1], galectin-2 [NP_598283.1], galectin-3 [NP_114020.1] and galectin-7 [NP_072104.2]), Horse (<i>Equus caballus</i>: galectin-1 [XP_001501082.2], galectin-2 [XP_001499566.2], galectin-3 [XP_005605252.1] and galectin-7 [XP_001496714.2]), Rhinoceros (<i>Ceratotherium simum simum</i>: galectin-1 [XP_004418181.1], galectin-2 [XP_004418176.1] and galectin-7 [XP_004441551.1]), Cow (<i>Bos taurus</i>: galectin-1 [NP_786976.1], galectin-2 [NP_001244020.1], galectin-3 [NP_001095811.1] and galectin-7 [XP_002695023.2]), Dog (<i>Canis lupus familiaris</i>: galectin-1 [ADR80617.1], galectin-2 [NP_001271396.1], galectin-3 [NP_001183972.1] and galectin-7 [NP_001183972.1]), Cat (<i>Felis catus</i>: galectin-1 [XP_003989294.1], galectin-2 [XP_006934157.1], galectin-3 [XP_003987704.1] and galectin-7 [XP_003987704.1]), Sheep (<i>Ovis aries</i>: galectin-1 [AAT38511.1], galectin-2 [XP_004007664.1], galectin-3 [XP_004010713.1] and galectin-7 [XP_004010713.1]), Pig (<i>Sus scrofa</i>: galectin-1 [NP_001001867.1], galectin-3 [NP_001090970.1] and galectin-7 [NP_001136315.1]) and Mouse (<i>Mus Musculus</i>: galectin-2 [NP_079898.2]).</p

Design, Synthesis, and Biochemical Evaluation of Phosphonate and Phosphonamidate Analogs of Glutathionylspermidine as Inhibitors of Glutathionylspermidine Synthetase/Amidase from <i>Escherichia coli</i>

Three phosphapeptides designed to mimic two distinct tetrahedral intermediates formed during either the synthesis or hydrolysis of glutathionylspermidine (Gsp) were synthesized and evaluated as inhibitors of the bifunctional enzyme Gsp synthetase/amidase. While the polyamine-containing phosphapeptides were determined to be potent and selective inhibitors, they selectively inhibit the synthetase activity over the amidase domain. A phosphonate-containing tetrahedral mimic is a reversible mixed-type inhibitor of Gsp synthetase with an inhibition constant of 6 μM for the inhibitor binding to the free enzyme (Ki) and 14 μM for the inhibitor binding to the enzyme−substrate complex (Ki‘). The corresponding phosphonamidate is a slow-binding inhibitor with a Ki of 24 μM and a Ki* (isomerization inhibition constant) of 0.88 μM. A non-polyamine-containing phosphonamidate exhibits no significant inhibition of the synthetase or amidase activity

Water-mediated interactions of hGal1 and hGal3 with the LN2 molecules.

<p>(A and B) Stereoview of LN2 molecule bound in the carbohydrate-recognition site of hGal1 (PDB ID: 1W6P) and hGal3-CRD (1KJL), respectively. LN2 ligand is depicted as gray stick model. The water (blue sphere) is coordinated by the H-bonds (green dashes) from N2 atom of LN2 and amino acid residues of galectins. <i>2F</i>_<i>o</i><i>-F</i>_<i>c</i> omit electron density (gray mesh) of the water molecules are highlighted and contoured at 1σ. Unique salt bridge network of hGal1 and hGal3-CRD are indicated as yellow dashes. (C and D) Structural superposition of LN1 and LN2 complex structures from hGal1 (C) and hGal3-CRD (D). The C5-hydroxyl group of LN1 (yellow sticks) in hGal1 and hGal3-CRD complexes makes close contact with the coordinated water in LN2-hGal1 andLN2-hGal3-CRD complex with a distance of 2.2 and 2.1 Å, respectively. (E) Structure of hGal7 (shown in color green) in complex with LN2 (orange) is superimposed with LN2-hGal3-CRD complex structure (all in gray). Most LN2-contacting residues in hGal7 (such as Arg53^hGal7, Trp69^hGal7, Glu72^hGal7 and Arg74^hGal7) are well superimposed with those of hGal3, except the Glu58^hGal7 residue. Location of Glu58^hGal7 is distinctive from Glu165^hGal3/ Asp54^hGal1 and distance between Glu58^hGal7 and N2 atom of LN2 is too far for them to coordinate a water molecule in between.</p

Structural Basis Underlying the Binding Preference of Human Galectins-1, -3 and -7 for Galβ1-3/4GlcNAc

<div><p>Galectins represent β-galactoside-binding proteins and are known to bind Galβ1-3/4GlcNAc disaccharides (abbreviated as LN1 and LN2, respectively). Despite high sequence and structural homology shared by the carbohydrate recognition domain (CRD) of all galectin members, how each galectin displays different sugar-binding specificity still remains ambiguous. Herein we provided the first structural evidence of human galectins-1, 3-CRD and 7 in complex with LN1. Galectins-1 and 3 were shown to have higher affinity for LN2 than for LN1, while galectin-7 displayed the reversed specificity. In comparison with the previous LN2-complexed structures, the results indicated that the average glycosidic torsion angle of galectin-bound LN1 (ψ^LN1 ≈ 135°) was significantly differed from that of galectin-bound LN2 (ψ^LN2 ≈ -108°), i.e. the GlcNAc moiety adopted a different orientation to maintain essential interactions. Furthermore, we also identified an Arg-Asp/Glu-Glu-Arg salt-bridge network and the corresponding loop (to position the second Asp/Glu residue) critical for the LN1/2-binding preference.</p></div

Data_Sheet_1_The Incidence of Contrast-Induced Nephropathy and the Need of Dialysis in Patients Receiving Angiography: A Systematic Review and Meta-Analysis.docx

ObjectivesThe risk of dialysis following contrast exposure is unclear. We aimed to examine the overall risk of contrast induced nephropathy and the need of dialysis based on a systematic review with random-effects meta-analysis.MethodsWe searched the electronic database including PubMed, Medline, Embase, and Cochrane Library from inception to 31 October, 2020 with predetermined search term to identify relevant studies. Observational studies investigating the association between contrast induced nephropathy after angiography and the need of dialysis were included, and summary risks were estimated. Two independent reviewers extracted the data, followed with random effects model to calculate the overall pooled incidence of contrast induced nephropathy and the need of dialysis after angiography. Subgroup-analysis and meta-regression were performed to assess heterogeneity of incidence across studies.ResultsOf 2,243 identified articles, 259 met our inclusion criteria were included in the meta-analysis after screening. Pooled effect estimates had the following summary incidence proportion for contrast induced nephropathy after angiography: 9.06% (95% CI: 8.53–9.58%; derived from 120 studies) and 0.52% (95% CI: 0.37–0.70%; derived from 110 studies) for the need of dialysis, respectively. The stratified summary incidence proportion of contrast induced nephropathy after contrast administration via intra-arterial route was 9.60% (95% CI: 9.0–10.2%; derived from 106 studies) and was 0.6% (95% CI: 0.40–0.80%; derived from 100 studies) for the need of dialysis, respectively. Our meta-regressions found that the amount of contrast medium exposure was associated with contrast-induced nephropathy.ConclusionThe potential risk of dialysis needs to be communicated to patients undergoing procedures requiring contrast, especially via intra-arterial exposure.Systematic Review Registration[https://reurl.cc/8Wrlry], identifier [CRD42020170702].</p

Data collection and refinement statistics.

<p>¹Statistics for data from the highest-resolution shell are shown in parentheses.</p><p>²</p><p></p><p></p><p></p><p><mi>R</mi></p><p><mi>s</mi><mi>y</mi><mi>m</mi></p><p></p><mo>=</mo><p><mo>(</mo></p><p><mi>Σ</mi><mi>Σ</mi></p><p><mo>|</mo></p><p></p><p><mi>I</mi></p><p><mi>h</mi><mi>k</mi><mi>l</mi></p><p></p><mo>−</mo><p><mo>〈</mo><mi>I</mi><mo>〉</mo></p><p></p><mo>|</mo><p></p><p></p><mo>)</mo><p></p><mo>/</mo><p><mo>(</mo></p><p><mi>Σ</mi></p><p><mi>I</mi></p><p><mi>h</mi><mi>k</mi><mi>l</mi></p><p></p><p></p><mo>)</mo><p></p><p></p><p></p><p></p>, where the average intensity <<i>I</i>> is taken overall symmetry equivalent measurements and <i>I</i>_<i>hkl</i> is the measured intensity for any given reflection.<p></p><p>³</p><p></p><p></p><p></p><p><mi>R</mi></p><p><mi>w</mi><mi>o</mi><mi>r</mi><mi>k</mi></p><p></p><mo>=</mo><p><mo>(</mo></p><p><mi>Σ</mi></p><p><mo>|</mo></p><p></p><p><mo>|</mo></p><p></p><p><mi>F</mi><mi>o</mi></p><p></p><mo>|</mo><p></p><mo>−</mo><mi>k</mi><p><mo>|</mo></p><p></p><p><mi>F</mi><mi>c</mi></p><p></p><mo>|</mo><p></p><p></p><mo>|</mo><p></p><p></p><mo>)</mo><p></p><mo>/</mo><p><mo>(</mo></p><p><mi>Σ</mi></p><p><mo>|</mo></p><p></p><p><mi>F</mi><mi>o</mi></p><p></p><mo>|</mo><p></p><p></p><mo>)</mo><p></p><p></p><p></p><p></p>, where <i>F</i>_<i>o</i> and <i>F</i>_<i>c</i> are the observed and calculated structure factor amplitudes, respectively.<p></p><p>⁴<i>R</i>_<i>free</i> was calculated for R factor using only an unrefined subset of reflections data (5%).</p><p>Data collection and refinement statistics.</p

Structural comparisons among hGal1, 3-CRD and 7.

<p>(A) S4-S6 β-strands of hGal1 (pink), 3-CRD (blue) and 7 (green) are superimposed. The unique salt bridge network of hGal1 (R48-D54-E71-R73), 3-CRD (R162-E165-E184-R186) and 7 (R53-E58-E72-R74) are shown in stick models. (B) Structure-based sequence alignment of S4-S6 β-strands of hGal1, 3-CRD and 7. Secondary structures were designated according to the resolved x-ray structures. The highly conserved LNs-interacting residues among hGal1, 3-CRD and 7 are indicated by asterisks. Residues involved in unique salt bridge network of hGal1, 3-CRD and 7 are colored in either red (Glu/Asp) or blue (Arg).</p