Highly sensitive and selective detection of PCB 77 using an aptamer-catalytic hairpin assembly in an aquatic environment

Polychlorinated biphenyls (PCBs) are synthetic organic compounds that are extremely difficult to break down in water and can accumulate in human fat and organisms. However, methods that can be used to detect large amounts of PCBs remain unsatisfactory, as they are generally overly sensitive and involve complex operations. An aptamer-based catalytic hairpin assembly (aptamer-CHA) reaction for the selective detection of 3,3′,4,4′-tetrachlorobiphenyl (PCB 77) was developed. It combines the advantages of aptamers and signal amplification reactions. The aptamer selectivity recognizes the target, PCB 77, which triggers the sensitive CHA reaction to produce a fluorescence signal. CHA is a sensitive enzyme-free signal amplification method suitable for on-site detection. Therefore, the identification aptamer is the basis for the quantitative detection of PCB 77, with a detection range of 0.01 μg L⁻¹ to 500 μg L⁻¹ and a detection limit of 0.01 μg L⁻¹. In this study, the aptamer was used to improve the selectivity of the reaction, and the CHA reaction improved the sensitivity of the detection system. Such high-sensitivity PCB detection capabilities with simplified procedures may be useful for real-time field detection and other monitoring tasks. This method can be used as a rapid fluorescence detection strategy for other targets in aquatic environments.

The fluorescence sensing strategy was used to detect PCB77 based on the aptamer-complex and catalytic hairpin assembly.     

A label-free RTP sensor based on aptamer/quantum dot nanocomposites for cytochrome c detection

Given the outstanding room-temperature phosphorescence (RTP) of Mn–ZnS quantum dots (QDs) and the specific recognition performance of the aptamer, we built phosphorescent composites from aptamers conjugated with polyethyleneimine quantum dots (PEI-QDs) and applied them to cytochrome c (Cyt c) detection. Specifically, QDs/CBA composites were generated from the electrostatic interaction between the positively-charged PEI-QDs and the negatively-charged Cyt c binding aptamer (CBA). With the presence of Cyt c, the Cyt c can specifically bind with the QDs/CBA composites, and quench the RTP of QDs through photoinduced electron-transfer (PIET). Thereby, an optical biosensor for Cyt c detection was built, which had a detection range of 0.166–9.96 μM and a detection limit of 0.084 μM. This aptamer-mediated phosphorescent sensor with high specificity and operational simplicity can effectively avoid the interference of scattering light from complex substrates. Our findings offer a new clue for building biosensors based on QDs and aptamers.

In this study, the nanocomposites from polyethyleneimine-capped Mn-doped ZnS QDs (PEI-QDs) and Cyt c binding aptamer (CBA) were prepared and used as Cyt c RTP sensors..     

Highly photoluminescent and temperature-sensitive P, N,B-co-doped carbon quantum dots and their highly sensitive recognition for curcumin

Water-soluble P, N, B-co-doped carbon quantum dots (PNBCDs) synthesized using a convenient hydrothermal method exhibit many excellent features, such as strong fluorescence, excitation-independent emission, high monodispersity, good stability, and excellent water solubility with a fluorescence quantum yield of 21.95%. The as-prepared PNBCDs possessed remarkable selectivity and sensitivity towards curcumin with the linear range of 0–1.5 μmol L⁻¹ and the detection limit for curcumin was 68 nmol L⁻¹ (3σ/k). Additionally, the wonderfully reversible and repeatable sensitivity to external temperature makes it possible that the PNBCDs could be used as a biocompatible fluorescent ink and for thermo-sensitive devices.

Temperature-sensitive P, N, B-co-doped carbon quantum dots (PNBCDs) synthesized using one-pot method exhibit many excellent features, such as strong fluorescence, good stability and sensitive detection for curcumin.     

A fluorescent aptasensor based on gold nanoparticles quenching the fluorescence of rhodamine B to detect acetamiprid

Pesticide residue detection is one of the main safety issues in the utilization of medicinal plants. In this work, a highly selective and sensitive aptasensor for acetamiprid determination was designed. The mechanism of the proposed method is based on the fluorescence resonance energy transfer (FRET) between gold nanoparticles (AuNPs) and rhodamine B (RB). Aptamers protect AuNPs from salt-induced aggregation, which causes fluorescence quenching of RB by the AuNPs via surface energy transfer. In the absence of acetamiprid, AuNPs were coated with aptamers on the surface and dispersed in NaCl solution. At this time, the dispersed AuNPs could perfectly quench the fluorescence intensity of RB. In contrast, in the presence of acetamiprid, aptamers specifically combine with acetamiprid to form a complex. With a high salt concentration, AuNPs would be aggregated without aptamer protection, weakening the RB quenching effect. Therefore, the concentration of acetamiprid could be obtained from the change in fluorescence intensity in the system. A fluorescent sensing method was established with a linear range from 0.1 to 3 μg mL⁻¹, and the LOD was 0.0285 μg mL⁻¹. The recoveries of acetamiprid in traditional Chinese medicine (TCM) samples were 96.23–105.75%. This method has great application value for the detection of acetamiprid in a complex sample matrix.

In this paper, we present a turn-on fluorescent approach to detect acetamiprid in traditional Chinese medicine, with a high sensitivity and low detection limit. It provides more application possibilities for fluorescence-aptamer biosensing.     

The use of S2O82− and H2O2 as novel specific masking agents for highly selective “turn-on” fluorescent switching recognition of CN− and I− based on Hg2+–graphene quantum dots

In this study, we report that both CN⁻ and I⁻ can enhance the fluorescent intensity of Hg²⁺–graphene quantum dots (Hg²⁺–GQDs). However, the selectivity of the sensor was poor. Accordingly, simple specific masking agents can be directly used to solve this problem. Here, for the first time, we report the use of persulfate ion (S₂O₈²⁻) as a turn-on fluorescent probe of Hg²⁺–GQDs for selective CN⁻ detection, while hydrogen peroxide (H₂O₂) was selected for its sensing ability towards I⁻ ion detection. Interestingly, the signal was immediately measured after addition of the masking agent to Hg²⁺–GQDs and the sample because its interaction was very fast and efficient. The method had a linear response in the concentration ranges of 0.5–8 μM (R² = 0.9994) and 1–12 μM (R² = 0.9998) with detection limits of 0.17 and 0.20 μM for CN⁻ and I⁻, respectively. The sensor was successfully used for the dual detection of both CN⁻ and I⁻ in real water samples with satisfactory results. In conclusion, the specific masking agents in a Hg²⁺–GQDs system appeared to be good candidates for fluorometric “turn-on” sensors for CN⁻ and I⁻ with excellent selectivity over other ions.

In this study, we report that both CN⁻ and I⁻ can enhance the fluorescent intensity of Hg²⁺–graphene quantum dots (Hg²⁺–GQDs).     

Aptamer fluorescence anisotropy assays for detection of aflatoxin B1 and adenosine triphosphate using antibody to amplify signal change

Fluorescence polarization/anisotropy (FP/FA) is an attractive technology for determining small molecules in homogeneous solution based on rotation changes of a fluorescent reporter. Binding induced conformation change is a specific property of aptamers. This property has been integrated into aptamer based FA assays for small molecules. In this work, we reported aptamer FA assays for aflatoxin B1 (AFB1) and adenosine triphosphate (ATP) by using antibody conjugated complementary DNA at the 3′ end and a fluorescein (FAM)-labeled aptamer at the 5′ end. The hybridization of aptamer and cDNA induced a FAM label close to the large-sized antibody, which restricts the local rotation of FAM and gives high FA signal. With the addition of target, the aptamer probe binds with the target, and the aptamer–cDNA duplex is inhibited, causing FA signal decreases. This method achieved detection of 25 pM AFB1 and 1 μM ATP, respectively. The assay is promising for application.

Aptamer fluorescence anisotropy assays for small molecules (aflatoxin B1 and ATP) using antibody to amplify signal change.     

Facile synthesis of N,P-doped carbon dots from maize starch via a solvothermal approach for the highly sensitive detection of Fe3+

Nitrogen/phosphorus-doped carbon dots (N, P-CDs) with a quantum yield as high as 76.5% were synthesized by carbonizing maize starch via a facile ethanol solvothermal approach. Transmission electron microscopy (TEM) measurement shows that the as-prepared N, P-CDs displayed a quasi-spherical shape with a mean size of ca. 2.5 nm. Fourier transform infrared spectroscopy and X-ray photoelectron spectroscopy disclosed the presence of –OH, –NH₂, –COOH, and –CO functional groups over the surface of N, P-CDs. On the basis of excellent fluorescent properties with strong blue fluorescence emission at 445 nm upon excitation at 340 nm, these N, P-CDs were adopted as a fluorescent probe towards the effective detection of Fe³⁺ ions in water. The limit of detection (LOD) was as low as 0.1 μmol L⁻¹ and showed a better linear relationship in the range of 0.1 ∼ 50 μmol L⁻¹. In conclusion, these synthesized N, P-CDs can be efficiently used as a promising candidate for the detection of Fe³⁺ ions in some practical samples.

Nitrogen/phosphorus-doped carbon dots (N, P-CDs) with a quantum yield as high as 76.5% were synthesized by carbonizing maize starch via a facile ethanol solvothermal approach and utilized for the detection of Fe³⁺.     

Development of a fluorescence assay for highly sensitive detection of Pseudomonas aeruginosa based on an aptamer-carbon dots/graphene oxide system

An aptamer-based fluorescence assay for culture-independent detection of Pseudomonas aeruginosa was developed. This assay was enabled by highly specific aptamers conjugated with photoluminescent carbon dots (CDs) as the fluorescent probe and graphene oxide (GO) as the quencher. Specially, high-throughput sequencing was achieved during systematic evolution of ligands via exponential enrichment (SELEX) for accurate recognition of aptamers. This assay displayed high specificity towards P. aeruginosa and was resistant to interference by other ubiquitous bacteria including Escherichia coli, Bacillus subtilis, Staphylococcus aureus, Enterococcus faecalis, and Clostridium perfringens. After the conditions were optimized, this assay achieved a wide detection range for P. aeruginosa varying from 10¹ CFU mL⁻¹ to 10⁷ CFU mL⁻¹. Notably, it approached an excellent detection limit as low as 9 CFU mL⁻¹. Therefore, this fluorescence assay was considered successfully developed for highly sensitive detection of P. aeruginosa. This assay also detected the contamination of P. aeruginosa in tap water and commercial bottled water, thereby suggesting its potential application in real water samples.

The fluorescence assay for detecting Pseudomonas aeruginosa was achieved by the aptamer-CDs/GD system which exhibited obvious fluorescence recovery when encountered the cells of P. aeruginosa.     

Novel β-cyclodextrin doped carbon dots for host–guest recognition-assisted sensing of isoniazid and cell imaging

In the present study, novel β-cyclodextrin doped carbon dots (CCDs) were prepared via a simple one-pot hydrothermal method at a mild temperature (140 °C), using mixtures of β-cyclodextrin and citric acid as precursors. By characterizing the chemical properties of CCDs prepared at 140 °C and 180 °C, the importance of low-temperature reaction for preservation of the specific structure of β-CD was elucidated. The CCDs showed excellent optical properties and were stable to changes in pH, ionic strength and light irradiation. Since the fluorescence of the CCDs could be selectively quenched by isoniazid (INZ) through specific host–guest recognition effects, a convenient isoniazid fluorescence sensor was developed. Under the optimal conditions, the sensor exhibited a relatively low detection limit of 0.140 μg mL⁻¹ and a wide detection range from 0.2 μg mL⁻¹ to 50 μg mL⁻¹ for INZ detection. Furthermore, the sensor was employed successfully for the determination of INZ in urine samples with satisfactory recovery (91.1–109.5%), displaying potential in clinical applications. Finally, low cytotoxicity of the prepared CCDs was confirmed using the CCK-8 method, followed by application in HepG2 cell imaging.

Novel β-cyclodextrin (β-CD) doped carbon dots (CCDs) were prepared at a mild temperature to preserve the host–guest recognition properties of β-CD. An isoniazid fluorescence sensor was constructed with a limit of detection of 0.14 μg mL⁻¹.     

Enhanced field emission properties of graphene-based cathodes fabricated by ultrasonic atomization spray

Two types of graphene cathodes were constructed by doctor blade and ultrasonic atomization spray respectively. The effects of cathodic film morphology and thickness on graphene cathodes'' field emission performance were investigated. Ultrasonic atomization spray coated graphene cathodes possess a much lower turn-on and threshold field but much higher emission current density than graphene cathodes coated by doctor blade. The enhanced field emission properties can be ascribed to the suppression of field-screening effect by roughened surface geometry rendered by ultrasonic atomization spray. For ultrasonic atomization sprayed graphene cathodes at a film thickness of 116 μm, an emission current density as large as 29.6 mA cm⁻² was achieved at 5 V μm⁻¹, along with a turn-on field and threshold field as low as 1.52 V μm⁻¹ and 2.65 V μm⁻¹, respectively. Emission stability examination shows no visible emission current density fluctuation or decline over a 10 hour operation at ∼3.72 mA cm⁻², demonstrating excellent field emission stability for ultrasonic atomization sprayed graphene cathodes. The luminance test also indicates good uniformity of electron emission from ultrasonic atomization sprayed graphene cathodes. The above experiment results indicate that the ultrasonic atomization spray method is suitable for large-area fabrication of high-performance graphene field emitters and holds great potential for applications in field emission displays.

Ultrasonic atomization spray endowed graphene cathodes with roughened 3-D array-like surface geometry which greatly enhanced electron emission.     

Highly fluorescent carbon dots as an efficient nanoprobe for detection of clomifene citrate

Highly fluorescent carbon dots (CDs) were synthesized through facile hydrothermal carbonization and ethylenediamine passivation of an easily available prawn shell precursor. The as-prepared CDs exhibit high water solubility, wavelength-tunable fluorescence with quantum yield up to 68.9%, high photostability and resistance against biomolecules, thus enabling the application as viable fluorescent nanoprobes for detection of guest quenchers. The fluorescence of the CDs can be effectively quenched by clomifene citrate (CC, a common drug for infertility) through static quenching, and therefore can serve as a simple and efficient fluorescent nanoprobe for determination of CC with wide linear range (0.25–10 μg mL⁻¹) and low detection limit (0.2 μg mL⁻¹). The CDs also showed low cytotoxicity, which enables the safe and accurate fluorescent detection of spiked CC in human serum, demonstrating their potential as a credible fluorescent CC nanoprobe in clinical examination.

Highly fluorescent carbon dots (CDs) were synthesized through facile hydrothermal carbonization and ethylenediamine passivation of an easily available prawn shell precursor.     

A highly sensitive fluorescent immunosensor for sensitive detection of nuclear matrix protein 22 as biomarker for early stage diagnosis of bladder cancer

A novel strategy is reported for highly sensitive, rapid, and selective detection of nuclear matrix protein NMP22 using two-color quantum dots based on fluorescence resonance energy transfer (FRET). Quantum dots (QDs) are highly advantageous for biological imaging and analysis, particularly when combined with (FRET) properties of semiconductor quantum dot (QDs) are ideal for biological analysis to improve sensitivity and accuracy. In this FRET system narrowly dispersed green emitting quantum dot CdTe core is used as a donor and labelled by monoclonal (mAb) antibody, while orange emitting quantum dot CdTe/CdS core shell is used as an accepter and labelled by polyclonal (pAb) antibody. The quantum dots are labelled by antibodies using EDC/NHS as crosslinking agent. Bovine serum albumin (BSA) solution was added to block nonspecific binding sites. The fluorescence intensity of QDs accepter decreased linearly with the increasing concentrations of NMP22 from 2–22 pg mL⁻¹ due to FRET system and fluoroimmunoassay reaction. This method has good regression coefficient (R² = 0.998) and detection limit was 0.05 pg mL⁻¹. The proposed FRET-based immunosensor provides a quick, simple and sensitive immunoassay tool for protein detection, and can be considered as a promising approach for clinical applications. The proposed FRET-based immunosensor provides a quick, simple and sensitive immunoassay tool for protein detection, and can be considered as a promising approach for clinical applications.

A novel strategy is reported for highly sensitive, rapid, and selective detection of nuclear matrix protein NMP22 using two-color quantum dots based on fluorescence resonance energy transfer (FRET).     

Low-cost electrochemical detection of l-tyrosine using an rGO–Cu modified pencil graphite electrode and its surface orientation on a Ag electrode using an ex situ spectroelectrochemical method

A low cost reduced graphene oxide–copper hybrid nano thin-film modified Pencil Graphite Electrode has been employed to detect the l-tyrosine enantiomer. The free-standing rGO–Cu hybrid nano-thin film was prepared by a simple one-step liquid–liquid interface method. Electrochemical Cyclic Voltammetry, Differential Pulse Voltammetry, pH-dependent and scan rate dependent studies on bare PGE, Cu, rGO, and rGO–Cu for l-tyrosine have been explained in detail. The rGO–Cu modified PGE based biosensor exhibits good detection of l-tyrosine. The linear range detection limit was estimated to be 1 × 10⁻⁷ M. The calculated sensitivity is 0.4 μA ppm⁻¹ mm². This electroactive biosensor is easily fabricated and controlled and is cost-effective. The surface orientation of l-tyrosine on the Ag electrode at a particular potential and its comparison with vibrational DFT calculations have been studied for the first time.

A low cost reduced graphene oxide–copper hybrid nano thin-film modified pencil graphite electrode has been employed to detect the l-tyrosine enantiomer.     

Enzyme-free ultrasensitive fluorescence detection of epithelial cell adhesion molecules based on a toehold-aided DNA recycling amplification strategy

Epithelial cell adhesion molecules (EpCAMs) play a significant role in tumorigenesis and tumor development. EpCAMs are considered to be tumor signaling molecules for cancer diagnosis, prognosis and therapy. Herein, an enzyme-free and highly sensitive fluorescent biosensor, with a combined aptamer-based EpCAM recognition and toehold-aided DNA recycling amplification strategy, was developed for sensitive and specific fluorescence detection of EpCAMs. Due to highly specific binding between EpCAMs and corresponding aptamers, strand a, which is released from the complex of aptamer/strand a in the presence of EpCAMs which is bound to the corresponding aptamer, triggered the toehold-mediated strand displacement process. An amplified fluorescent signal was achieved by recycling strand a for ultrasensitive EpCAM detection with a detection limit as low as 0.1 ng mL⁻¹, which was comparable or superior to that of reported immunoassays and biosensor strategies. In addition, high selectivity towards EpCAMs was exhibited when other proteins were selected as control proteins. Finally, this strategy was successfully used for the ultrasensitive fluorescence detection of EpCAMs in human serum samples with satisfactory results. Importantly, the present strategy may be also expanded for the detection of other targets using the corresponding aptamers.

A fluorescent biosensor with a combined aptamer-based EpCAM recognition and toehold-aided DNA recycling amplification strategy was developed.     

Highly rapid and non-enzymatic detection of cholesterol based on carbon nitride quantum dots as fluorescent nanoprobes

In this work, we reported a highly rapid and non-enzymatic method for cholesterol measuring based on carbon nitride quantum dots (CNQDs) as fluorescent nanoprobes, which were synthesized through chemical oxidation. The obtained CNQDs displayed high quantum yield up to 35% as well as excellent photostability, water solubility and low toxicity. We found that the fluorescence of CNQDs could be quenched more than 90% within 30 seconds by cholesterol through the formation of hydrogen bonds between –NH₂, –NH on the surface of CNQDs and cholesterol containing –OH. According to this phenomenon, a cholesterol detection method was constructed with a wide linear region over the range of 0–500 μmol L⁻¹ and a detection limit as low as 10.93 μmol L⁻¹, and it possessed the obvious advantages of being a very rapid process and avoiding the use of enzymes. In addition, this method showed high selectivity in the presence of various interfering reagents and applicability to the measurement of cholesterol in fetal bovine serum, which indicated its potential application value in clinical settings.

Highly rapid and non-enzymatic method for the detection of cholesterol was constructed based on carbon nitride quantum dots (CNQDs) as fluorescent nanoprobes. The fluorescence of CNQDs could be effectively and rapidly quenched by cholesterol.     

Using dual exonucleases to finely distinguish structural adjustment of aptamers for small-molecule detection

The binding of small molecules to their DNA aptamers can modulate their susceptibility to digestion by exonucleases, however, absolute differentiation between digestion and inhibition has never been reported. Here, we show that the digestion of aptamers by T7 exonuclease can be completely inhibited upon binding of small-molecule targets and exploit this finding for the first time to achieve sensitive, label-free small-molecule detection. We use a quinine-binding aptamer to show that target binding entirely halts T7 exonuclease digestion, leaving behind an intact double-stranded product that retains strong target affinity. On the contrary, digestion of nontarget-bound aptamer produces a single-stranded product incapable of target binding. Exonuclease I efficiently eliminates these single-stranded products but is unable to digest the target-bound double-stranded product. The remaining products can be fluorescently quantified with SYBR Gold to determine target concentrations, giving a limit of detection of 100 nM with the linear range from 0 to 8 μM. We demonstrate the first example of a dual-exonuclease-mediated approach capable of producing a concentration-dependent response in terms of aptamer digestion modules, therefore improving performance of the current aptamer-based assay for small-molecule detection.

Dual exonucleases to finely distinguish structural adjustment of aptamers to produce absolute differentiation between digestion and inhibition.     

Co2+ detection,cell imaging,and temperature sensing based on excitation-independent green-fluorescent N-doped carbon dots

Green-fluorescent N-doped carbon dots (N-CDs) have been successfully fabricated using hydrothermal treatment of tyrosine and urea. The N-CDs obtained showed excitation-independent emission, superior stability and strong photoluminescence with a quantum yield of ca. 9.8%. Based on these striking behaviors, the as-prepared N-CDs have been utilized in Co²⁺ detection and temperature sensing. Due to an inner filter effect, the N-CDs obtained were dramatically quenched by Co²⁺ with linear ranges of 0.1 μM–10 μM, 25 μM–275 μM and 300 μM–400 μM, and they had a detection limit of 0.15 μM. The use of the as-prepared N-CDs has been extended to visualize Co²⁺ fluctuations in living cells. Additionally, the N-CDs obtained have also been applied for use as a temperature sensor with a linear range of 25–80 °C.

Green-fluorescent N-doped carbon dots (N-CDs) have been successfully fabricated using hydrothermal treatment of tyrosine and urea.     

MOF-derived (MoS2, γ-Fe2O3)/graphene Z-scheme photocatalysts with excellent activity for oxygen evolution under visible light irradiation

Constructing Z-scheme heterojunctions is considered as an effective strategy to obtain catalysts of high efficiency in electron–hole separation in photocatalysis. Unfortunately, suitable heterojunctions are difficult to fabricate because the direct interaction between two semiconductors may lead to unpredictable negative effects such as electron scattering or electron trapping due to the existence of defects which causes the formation of new substances. Furthermore, the van der Waals contact between two semiconductors also results in bad electron diffusion. In this work, a MOF-derived carbon material as a Z-scheme photocatalyst was synthesized via one-step thermal treatment of MoS₂ dots @Fe-MOF (MIL-101). Under visible light irradiation, the well-constructed Z-scheme (MoS₂, γ-Fe₂O₃)/graphene photocatalyst shows 2-fold photocatalytic oxygen evolution activity (4400 μmol g⁻¹ h⁻¹) compared to that of γ-Fe₂O₃/graphene (2053 μmol g⁻¹ h⁻¹). Based on ultraviolet photoelectron spectrometry (UPS), Mott–Schottky plot, photocurrent and photoluminescence spectroscopy (PL) results, the photo-induced electrons from the conduction band of γ-Fe₂O₃ could transport quickly to the valence band of MoS₂via highly conductive graphene as an electron transport channel, which could significantly enhance the electron–hole separation efficiency as well as photocatalytic performance.

The heterojunction between MoS₂ and γ-Fe₂O₃ was constructed via linking by in situ formed graphene, which resulted in a good photocatalyst for the oxygen evolution reaction, showing O₂ evolution activity of 4400 μmol g⁻¹ h⁻¹.     

Selection of potential aptamers for specific growth stage detection of Yersinia enterocolitica

Yersinia enterocolitica remains a threat to public health, and a sensitive detection method is a prerequisite due to its complicated diagnosis associated with slow growth. Recently, aptamer-based detection systems have played a vital role in the development of simple, rapid, sensitive, and specific detection methods. Herein, highly specific ssDNA aptamers were screened against Y. enterocolitica at the different growth stages by whole cell-SELEX. Cells at different growth stages were harvested and incubated with an ssDNA library to get an enriched pool of specific aptamer candidates. After the 10^th round of SELEX, the enriched pool was sequenced and grouped into seven families based on homology and similarity of the secondary structure. Flow cytometry analysis revealed that the aptamers M1, M5, and M7 with K_d values of 37.93 ± 7.88 nM, 74.96 ± 21.34 nM, and 73.02 ± 18.76 nM had the highest affinity and specificity to the target, respectively. The selected aptamers showed binding to the different growth stages of Y. enterocolitica with a significant increase in the gated fluorescence. Our aptamer selection strategy is convenient, and the developed aptamer can be useful for an accurate and reliable detection system.

Yersinia enterocolitica remains a threat to public health, and a sensitive detection method is a prerequisite due to its complicated diagnosis associated with slow growth.     

Branched-chain Amino Acid Nitrogen Transfer to Alanine In Vivo in Dogs: DIRECT ISOTOPIC DETERMINATION WITH [15N]LEUCINE

To investigate the contribution of branched-chain amino acids as a nitrogen source for alanine in vivo, dogs were infused with l-[¹⁵N]leucine, l-[U-¹⁴C]leucine, l-[2,3,3,3-²H₄]alanine, and d-[6,6-²H₂]-glucose. ¹⁴C and ¹⁵N isotopic equilibrium in plasma leucine, and deuterium enrichment in arterial and femoral plasma glucose and alanine were achieved within 3 h of initiation of the respective isotope infusion in all animals. The average flux of leucine determined by [¹⁵N]leucine was 5.4 μmol·kg⁻¹·min⁻¹, whereas using [¹⁴C]leucine it was 3.7 μmol·kg⁻¹·min⁻¹. Turnover rates for alanine and glucose were 11.0 and 17.2 μmol·kg⁻¹·min⁻¹, respectively.