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A new lateral flow immunoassay strip (LFIA) combining sensitive detection and identification of multiple bacteria remains a huge challenge. In this study, we first developed multifunctional urchin-shaped Au-Ag@Pt nanoparticles (UAA@P NPs) with a unique combination of colorimetric-SERS-photothermal-catalytic (CM/SERS/PT/CL) properties and integrated them with LFIA for multiplexed detection and specific discrimination of pathogenic bacteria in blood samples. Unlike the conventional LFIA that relied on antibody (Ab), this novel LFIA introduced 4-mercaptophenylboronic acid (4-MPBA) as an ideal Ab replacer that was functionalized on UAA@P NPs (UAA@P/M NPs) with outstanding binding and enrichment capacities toward bacteria. Taking Staphylococcus aureus ( S. aureus ) as model bacteria, the limit of detection (LOD) was 3 CFU/mL for SERS-LFIA, 27 CFU/mL for PT-LFIA, and 18 CFU/mL for CL-LFIA, three of which were over 330-fold, 37-fold, and 55-fold more sensitive than ordinary visual CM-LFIA, respectively. Besides, this SERS-LFIA is capable of identifying three types of bacterial spiked blood samples ( E. coli , S. aureus , and P. aeruginosa ) effectively according to specific bacterial Raman "fingerprints" by partial least-squares-discriminant analysis (PLS-DA). More importantly, this LFIA was successfully applied to blood samples with satisfactory recoveries from 90.3% to 108.8% and capable of identifying the infected patients ( N = 4) from healthy subjects ( N = 2) with great accuracy. Overall, the multimodal LFIA incorporates bacteria discrimination and quantitative detection, offering an avenue for early warning and diagnosis of bacterial infection.

To monitor the levels of mitochondrial DNA G-quadruplexes (mtDNA G4s) in spermatozoa and to explore the possibility using mtDNA G4s as a reliable marker in patients with multiple clinical insemination failures, a novel chemical TPE-mTO probe engineered in our previous work was used on both samples from the mice sperm and from patients with fertilization failure. Expression of valosin-containing protein and the zona-free hamster egg assay were used to evaluate mitophagy and human sperm penetration. RNA-sequencing was used to explore expression changes of key genes affected by mtDNA G4s. Results showed that the probe can track mtDNA G4s in spermatozoa easily and quickly with fewer backgrounds. Significantly increased mtDNA G4s were also found in patients with fertilization failure, using the flow-cytometry-based TPE-mTO probe detection method. A sperm-hamster egg penetration experiment showed that abnormal fertilization caused by increased mtDNA G4s can be effectively restored by a mitophagy inducer. This study provides a novel method for monitoring etiological biomarkers in patients with clinical infertility and treatment for patients with abnormal fertilization caused by mtDNA G4 dysfunction.

Harata-Kodaka's rule predicting the induced chirality of the guest molecules by cyclodextrins has been discovered in the 1970-1990s, yet its ability to control the supramolecular handedness of self-assembled structures has not been sufficiently recognized. Here we show that in a coordinating self-assembly system that is able to form racemic cone shells symmetry breaking occurs if the ligand is prethreaded into α-cyclodextrin prior to metal ion addition, and the handedness of cone shells can be rationally manipulated by creating the two scenarios of the Harata-Kadaka rule through controlling the host-guest dynamics. Since the coordination complexes have strong self-assembling ability, the coordinating ligand would dethread from the cavity of α-cyclodextrin but leaving the induced chirality to the coordinating self-assembly, thus catalyzing symmetry breaking. This work reveals that the dynamic factors such as concentration and molar ratio may play important roles in symmetry breaking at the supramolecular level. The current strategy provides a promising method for the symmetry breaking and manipulation of the handedness of self-assembled materials formed by achiral molecules.

Dissolved oxygen (DO) reflects river metabolic pulses and is an essential water quality measure. Our capabilities of forecasting DO however remain elusive. Water quality data, specifically DO data here, often have large gaps and sparse areal and temporal coverage. Earth surface and hydrometeorology data, on the other hand, have become largely available. Here we ask: can a Long Short-Term Memory (LSTM) model learn about river DO dynamics from sparse DO and intensive (daily) hydrometeorology data? We used CAMELS-chem, a new data set with DO concentrations from 236 minimally disturbed watersheds across the U.S. The model generally learns the theory of DO solubility and captures its decreasing trend with increasing water temperature. It exhibits the potential of predicting DO in "chemically ungauged basins", defined as basins without any measurements of DO and broadly water quality in general. The model however misses some DO peaks and troughs when in-stream biogeochemical processes become important. Surprisingly, the model does not perform better where more data are available. Instead, it performs better in basins with low variations of streamflow and DO, high runoff-ratio (>0.45), and winter precipitation peaks. Results here suggest that more data collections at DO peaks and troughs and in sparsely monitored areas are essential to overcome the issue of data scarcity, an outstanding challenge in the water quality community.

Intelligent control over the handedness of circularly polarized luminescence (CPL) is of special significance in smart optoelectronics, information storage, and data encryption; however, it still remains a great challenge to rationally design a CPL material that displays reversible handedness inversion without changing the system composition. Herein, we show this comes true by coupling the two scenarios of Harata-Kodaka's rule on the same supramolecular platform of crystalline microtubes self-assembled from surfactant-cyclodextrin host-guest complexes. Upon coassembling a linear dye with its electronic transition dipole moment outside of the cavity of β-CyD, the chirality transfer from the induced chirality of SDS in the SDS@2β-CyD microtubes to the dye generates left-handed CPL at room temperature. Upon elevating temperature, the dye forms inclusion complex with β-CyD, so that right-handed CPL is induced because the polar group of the dye is outside of the cavity of β-CyD. This process is completely reversible. We envision that host-guest chemistry would be very promising in creating smart CPL inversion materials for a vast number of applications., Competing Interests: The authors declare no competing financial interest., (© 2020 The Authors. Published by American Chemical Society.)

Eutrophication has threatened water resources worldwide, yet mechanistic understanding on controls of nutrient export remains elusive. This work tests the shallow and deep hypothesis: subsurface vertical chemical contrasts regulate nitrate export patterns under different land use conditions. We synthesized data from 228 watersheds and used reactive transport modeling (500 simulations) under broad land use, climate, and geology conditions. Data synthesis indicated that human perturbation has amplified chemical contrasts in shallow water (e.g., soil water) versus deep waters (e.g., groundwater), inducing primarily flushing patterns (concentrations increase with streamflow) in agriculture lands and dilution patterns (concentrations decrease with streamflow) in urban watersheds. Results revealed a quantitative relationship between export patterns and shallow-versus-deep concentration contrasts, underscoring the often-overlooked role of nutrient distribution over depth. Results challenge the commonly held perception that legacy stores in agricultural lands induce chemostasis where concentrations vary negligibly with streamflow. They suggest that nitrate concentrations from agricultural lands will escalate during large hydrological events, which can exacerbate nutrient export problems as flooding events intensify in the future climate.

Going beyond organic solvent as the solubilizer for small-molecular organic probes motivates exploration of water-soluble polymeric sensors. In this respect, dye-derived thermal-responsive polymeric sensors are an attractive direction, but for its practical application, it is limited by sensor recycling because only irreversible change in the structure of the recognition unit for many sensors can trigger the appearance of the detection signal. Here, we established the oligonucleotide-derived thermal-responsive polymeric sensor, TBC-P1, which overcame this fundamental limitation. The TBC-P1 sensor was based on reversible binding between oligonucleotides and Hg 2+ ions, and easy sensor separation via tuning temperature, achieving the Hg 2+ detection in a cost-effective and green manner. The TBC-P1 sensor displayed specific and rapid sensing properties toward Hg 2+ ions in pure aqueous media via turn-off fluorescence emission, with a limit of detection as low as 0.65 nM (much lower than the presently reported dye-derived polymeric sensors). This high detection sensitivity was further enhanced (with LOD = 0.17 nM) via warming to yield spherical micelles, in which the oligonucleotide-containing thermoresponsive PNIPAM block forming a hydrophobic core amplified the fluorescence signals. Treating the Hg 2+ -trapped micelles with cysteine (Cys) led to competition-induced release of these combined Hg 2+ ions and then thermally precipitating and recycling polymeric sensor TBC-P1. This oligonucleotide-derived thermalresponsive polymeric sensor will open a universal avenue for sensor recycling, which will achieve the goal of reducing cost and improving detection sensitivity of sensors.

The goal of this study is to investigate the biological response of mussel-inspired calcium phosphate cement (CPC) in vivo. Polydopamine (PDA), which is analogous to that of mussel adhesive proteins, was added in CPC. PDA-CPC was implanted into the femur, muscle, and critical-sized calvarial bone defects of rabbits. Histomorphometry of the sequential fluorescence sections showed that PDA-CPC was capable of forming more newborn bone than the control-CPC. More new bone, bone marrow cavity, and blood vessel were observed in PDA-CPC than in the control-CPC in decalcified and undecalcified histological sections. Necrosis bone was not observed in PDA-CPC, whereas it appeared in the control-CPC after 2 weeks. The histological sections in muscle witnessed that there was more ingrowth of collagen in PDA-CPC than that in the control-CPC. There were no significantly difference in the number of leukocyte between PDA-CPC and the control-CPC in blood. It was confirmed that the addition of PDA enhanced the bone repairing ability and biocompatibility of PDA-CPC. Push-out testing indicated that PDA increased the bonding strength between PDA-CPC and host bone in the early stage. These present results indicated that PDA-CPC might be one potential bone graft with gratifying biocompatibility and enhanced bone repairing.

The first palladium-catalyzed diastereoselective conjugate addition of arylboronic acids to chiral imides is reported. The catalytic system employing 4-tert-butyloxazolidin-2-one as the chiral auxiliary in a mixed solvent system of MeOH/H 2 O (1:3) under an air atmosphere provides the optically active 3-arylbutanoic acid derivatives in excellent yields with high diastereoselectivity.

Tidal flow constructed wetlands (TF CWs) have recently been studied as a sustainable technology to achieve enhanced nitrogen removal; however, the underlying mechanisms responsible for removing ammonium (NH4(+)) and nitrate (NO3(-)) have not been compared and quantified at the molecular level (genes) in controlled TF CWs. In this study, two TF CWs T1 (treating NH4(+) wastewater) and T2 (treating NO3(-) wastewater) achieved high removal efficiencies for chemical oxygen demand (COD, 92 ± 2.7% and 95 ± 2.4%, respectively), NH4(+)/NO3(-) (76 ± 3.9% and 97 ± 2.2%, respectively), and total nitrogen (TN, 81 ± 3.5% and 93 ± 2.3%, respectively). Combined analyses revealed that the presence of simultaneous nitrification, anammox, and denitrification processes and the coupling of dissimilatory nitrate reduction to ammonium, ammonia oxidation, and anammox were the primary reason accounted for the robust treatment performance in T1 and T2, respectively. Results from stepwise regression analysis suggested that the NH4(+) removal rate in T1 was collectively controlled by amoA, nxrA, and anammox, while the NO3(-) removal rate in T2 was governed by nxrA and narG gene.

Lead (Pb) is a prominent toxic metal in natural and engineered systems. Current knowledge on Pb toxicity to the activated sludge has been limited to short-term (≤24 h) toxicity. The effect of extended Pb exposure on process performance, bacterial viability, and community compositions remains unknown. We quantified the 24-h and 7-day Pb toxicity to chemical oxygen demand (COD) and NH3–N removal, bacterial viability, and community compositions using lab-scale experiments. Our results showed that 7-day toxicity was significantly higher than the short-term 24-h toxicity. Ammonia-oxidizing bacteria were more susceptible than the heterotrophs to Pb toxicity. The specific oxygen uptake rate responded quickly to Pb addition and could serve as a rapid indicator for detecting Pb pollutions. Microbial viability decreased linearly with the amount of added Pb at extended exposure. The bacterial community diversity was markedly reduced with elevated Pb concentrations. Surface analysis suggested that the adsorbed form of Pb could have contributed to its toxicity along with the dissolved form. Our study provides for the first time a systematic investigation of the effect of extended exposure of Pb on the performance and microbiology of aerobic treatment processes, and it indicates that long-term Pb toxicity has been underappreciated by previous studies.

Research on incorporation of both growth factors and silver (Ag) into hydroxyapatite (HA) coatings on metallic implant surfaces for enhancing osteoinductivity and antibacterial properties is a challenging work. Generally, Ag nanoparticles are easy to agglomerate and lead to a large increase in local Ag concentration, which could potentially affect cell activity. On the other hand, growth factors immobilization requires mild processing conditions so as to maintain their activities. In this study, bone morphology protein-2 (BMP-2) and Ag nanoparticle contained HA coatings were prepared on Ti surfaces by combining electrochemical deposition (ED) of Ag and electrostatic immobilization of BMP-2. During the ED process, chitosan (CS) was selected as the stabilizing agent to chelate Ag ions and generate Ag nanoparticles that are uniformly distributed in the coatings. CS also reduces Ag toxicity while retaining its antibacterial activity. Afterwards, a BMP/heparin solution was absorbed on the CS/Ag/HA coatings. Consequently, BMP-2 was immobilized on the coatings by the electrostatic attraction between CS, heparin, and BMP-2. Sustained release of BMP-2 and Ag ions from HA coatings was successfully demonstrated for a long period. Results of antibacterial tests indicate that the CS/Ag/HA coatings have high antibacterial properties against both Staphylococcus epidermidis and Escherichia coli. Osteoblasts (OB) culture reveals that the CS/Ag/HA coatings exhibit good biocompatibility. Bone marrow stromal cells (BMSCs) culture indicates that the BMP/CS/Ag/HA coatings have good osteoinductivity and promote the differentiation of BMSCs. Ti bars with BMP/CS/Ag/HA coatings were implanted into the femur of rabbits to evaluate the osteoinductivity of the coatings. Results indicate that BMP/CS/Ag/HA coatings favor bone formation in vivo. In summary, this study presents a convenient and effective method for the incorporation of growth factors and antibacterial agents into HA coatings. This method can be utilized to modify a variety of metallic implant surfaces.

This paper reports an ultrasensitive resonance scattering (RS) method to detect tetracycline (TET) in milk based on the competition of aptamers between nanogold and TET, aggregation of naked nanogold, nanocatalytic Fehling reaction, and RS signals of catalytic product Cu₂O cubic. The detection principle was confirmed by the nanoparticle size analyzer (NANOS), scanning electron microscopy (SEM), and X-ray diffraction (XRD). The variations of RS intensity had good linear correlation with TET concentrations, and the limit of detection was calculated as 11.6 nM. The proposed method was successfully applied for analysis of TET in milk, with total recoveries ranging from 105 to 109%.

To evaluate the accuracy ofisoelectric point determination by capillary isoelectric focusing, the pI values of nine proteins and a peptide, the pI values of which had been determined by other methods and ranging pI 3.55-9.60, were determined by capillary isoelectric focusing by cofocusing of recently developed peptide pI markers ranging 3.38-10.17, and the consistency of the pI values was examined. Isoelectric focusing was carried out in neutral polymer-coated capillaries, and the pH gradient was mobilized by pressure toward the cathode, to detect samples with absorption at 280 nm at a fixed detection point. Carrier ampholytes from two different suppliers and in different pH ranges were used. The sharp peaks of the highly pure peptide pI markers greatly facilitated the unambiguous identification of the peaks. When a carrier ampholyte ranging over the acidic side was used, the detection of acidic pI samples was anomalously delayed. This could be partly mitigated by reducing the viscosity of the anode solution in comparison with the pH gradient formed in the capillary. Since the detection times vs the pH relationships were not linear in most cases, the use of a linear calibration line over an entire pH gradient would be erroneous. Instead, the pI values of samples were calculated by assuming a linear relation for pH against detection time between two flanking marker peptides. Close agreement between the pI values, determined by capillary isoelectric focusing, and the reference values of the samples was observed within an average difference range of 0.04-0.08 pH unit with a sample consumption of 10-100 ng within 30-60 min. Some carrier ampholytes were preferentially more effective at either the acidic or the basic side of the pH gradient. For confirmation of the completion of focusing, the use of two different focusing times is recommended.

We have investigated the in vitro reconstitution of the 24-meric inner core domain (E2c) of the transacylase (E2) component of bovine branched-chain alpha-keto acid dehydrogenase complex. The yield of recombinant E2c (amino acid residues 161-421 of bovine E2) expressed in Escherichia coli was markedly increased by fusing the bacterial maltose-binding protein (MBP) to the amino terminus of bovine E2c. Following factor Xa digestion to remove the MBP moiety, E2c was completely unfolded in 4.5 M guanidine HCl (Gdn.HCl). The denatured E2c monomers (apparent M(r) = 27,000) were diluted 100-fold at 25 degrees C into a refolding buffer containing 5 mM Mg-ATP and a 4-fold molar excess of chaperonins GroEL and GroES. Full E2 activity was recovered in 45 min. Omission of the chaperonins in the refolding buffer failed to recover any E2 activity. Recovery of E2 activity obeyed hyperbolic kinetics as a function of the chaperonin-to-E2c molar ratio and showed a requirement for hydrolysis of Mg-ATP. A stable GroEL-E2c complex was isolated which, in the presence of GroES and Mg-ATP, generated active E2c 24-mers. Dissociation of recombinant E2c 24-mers into active trimers was achieved by incubation in 1.5 M Gdn.HCl at 25 degrees C. The E2c trimers with an apparent M(r) of 84,000 were isolated by sucrose density gradient centrifugation in the presence of the chaotropic reagent. Removal of 1.5 M Gdn.HCl resulted in the spontaneous reassembly of trimers into the native 24-mer structure independent of chaperonins.(ABSTRACT TRUNCATED AT 250 WORDS)

The conformational stabilities of native pig citrate synthase (PCS), a recombinant wild-type PCS, and six active-site mutant pig citrate synthases were studied in thermal denaturation experiments by circular dichroism and in urea denaturation experiments by using DTNB to measure the appearance of latent SH groups. His274 and Asp375 are conserved active-site residues in pig citrate synthase that bind to substrates and are implicated in the catalytic mechanism of the enzyme. By site-directed mutagenesis, His274 was replaced with Gly and Arg, while Asp375 was replaced with Gly, Asn, Glu, or Gln. These modifications were previously shown to result in 10(3)-10(4)-fold reductions in enzyme specific activities. The thermal unfolding of pig citrate synthase and the six mutants in the presence and absence of substrates showed large differences in the thermal stabilities of mutant proteins compared to the wild-type pig citrate synthase. The functions of His274 and Asp375 in ligand binding were measured by oxalacetate protection against urea denaturation. These data indicate that active-site mutations that decrease the specific activity of pig citrate synthase also cause an increase in the conformational stability of the protein. These results suggest that specific electrostatic interactions in the active site of citrate synthase are important in the catalytic mechanism in the chemical transformations as well as the conformational flexibility of the protein, both of which are important for the overall catalytic efficiency of the enzyme.

Two amino acid residues, His274 and Asp375, were replaced singly in the active site of pig citrate synthase (PCS) with Gly274, Arg274, Gly375, Asn375, Glu375, and Gln375. The nonmutant protein and the mutant proteins were expressed in and purified from Escherichia coli, and the effects of these amino acid substitutions on the overall reaction rate and conformation of the PCS protein were studied by initial velocity and full time course kinetic analysis, behavior during affinity column chromatography, and monoclonal antibody reactivity. Native and mutant proteins purified similarly had a subunit molecular weight of 50,000 and were homologous when examined with 10 independent a-PCS monoclonal IgGs or with a polyclonal anti-PHCS serum. No activity was detected for Asn375 or Gln375. The kcats of the other purified mutant proteins, however, were decreased by about 10(3) compared to the nonmutant enzyme activity. The Km for oxalacetate was decreased 10-fold in the Glu375 protein and was reduced by half in Gly274 and Arg274 PCSs, while the Km for acetyl-CoA was decreased 2-3-fold in Gly274, Arg274, and Gln375 PCSs. A mechanism is proposed that electrostatically links His274 and Asp375.