Development of a Novel Benzimidazole-Based Probe and Portable Fluorimeter for the Detection of Cysteine in Human Urine.

The measurement of cysteine in human urine and live cells is crucial for evaluating biological metabolism, monitoring and maintaining the immune system, preventing tissue/DNA damage caused by free radicals, preventing autoimmune diseases, and diagnosing disorders such as cystinuria and cancer. A method that uses a fluorescence turn-on probe and a portable fluorescence spectrometer device are crucial for highly sensitive, simple, rapid, and inexpensive cysteine detection. Herein, we present the synthesis and application of a benzimidazole-based fluorescent probe (ABIA) along with the design and development of a portable fluorescence spectrometer device (CysDDev) for detecting cysteine in simulated human urine. ABIA showed excellent selectivity and sensitivity in detecting cysteine over homocysteine, glutathione, and other amino acids with the response time of 1 min and demonstrated a detection limit of 16.3 nM using the developed CysDDev. Further, ABIA also demonstrated its utility in detecting intracellular cysteine, making it an excellent probe for bio-imaging assay.

Construction of self-enhanced photoelectrochemical platform for L-cysteine detection via electron donor-acceptor type coumarin 545 aggregates.

Self-enhanced electron donor-acceptor type coumarin 545 aggregates prepared via an anionic surfactant-assisted reprecipitation method provide an underlying approach for the photoelectrochemical detection of L-cysteine, which can be employed in aqueous solution without the addition of electron donors.

The Impact of Diet on Urinary Risk Factors for Cystine Stone Formation.

Despite the importance of dietary management of cystinuria, data on the contribution of diet to urinary risk factors for cystine stone formation are limited. Studies on the physiological effects of diet on urinary cystine and cysteine excretion are lacking. Accordingly, 10 healthy men received three standardized diets for a period of five days each and collected daily 24 h urine. The Western-type diet (WD; 95 g/day protein) corresponded to usual dietary habits, whereas the mixed diet (MD; 65 g/day protein) and lacto-ovo-vegetarian diet (VD; 65 g/day protein) were calculated according to dietary reference intakes. With intake of the VD, urinary cystine and cysteine excretion decreased by 22 and 15%, respectively, compared to the WD, although the differences were not statistically significant. Urine pH was significantly highest on the VD. Regression analysis showed that urinary phosphate was significantly associated with cystine excretion, while urinary sulfate was a predictor of cysteine excretion. Neither urinary cystine nor cysteine excretion was affected by dietary sodium intake. A lacto-ovo-vegetarian diet is particularly suitable for the dietary treatment of cystinuria, since the additional alkali load may reduce the amount of required alkalizing agents.

A vinyl sulfone clicked carbon dot-engineered microfluidic paper-based analytical device for fluorometric determination of biothiols.

A microfluidic paper-based analytical device integrating carbon dot (CDs) is fabricated and used for a fluorometric off-on assay of biothiols. Vinyl sulfone (VS) click immobilization of carbon dots (CDs) on paper was accomplished by a one-pot simplified protocol that uses divinyl sulfone (DVS) as a homobifunctional reagent. This reagent mediated both the click oxa-Michael addition to the hydroxyl groups of cellulose and ulterior covalent grafting of the resulting VS paper to NH2-functionalized CDs by means of click aza-Michael addition. The resulting cellulose nanocomposite was used to engineer an inexpensive and robust microfluidic paper-based analytical device (µPAD) that is used for a reaction-based off-on fluorometric assay of biothiols (GSH, Cys, and Hcy). The intrinsic blue fluorescence of CDs (with excitation/emission maxima at 365/450 nm) is turned off via the heavy atom effect of an introduced iodo group. Fluorescence is turned on again due to the displacement of iodine by reaction with a biothiol. The increase in fluorescence is related to the concentration over a wide range (1 to 200 µM for GSH and 5-200 µM for Cys and Hcy, respectively), and the assay exhibits a low detection limit (0.3 µM for GSH and Cys and 0.4 µM for Hcy). The method allows for rapid screening and can also be used in combination with a digital camera readout. Graphical abstract Schematic representation of a µPAD based on click immobilized carbon dots and used for a reaction-based fluorometric off-on assay of biothiols. The intrinsic blue fluorescence of carbon dots is turned off via the heavy atom effect of an introduced iodo group and turned on by the displacement of this atom by reaction with a biothiol.

Dopamine functionalized CuO nanoparticles: A high valued "turn on" colorimetric biosensor for detecting cysteine in human serum and urine samples.

An attempt has been made to design one step synthesis of dopamine coated copper oxide nanoparticles (CuO@DOP NPs) by using microwave radiation method. The luminescent properties of CuO@DOP NPs have been explored for making colorimetric and visual biosensor for L-cys. Natural occurring dopamine has used as a precursor for the coating of CuO NPs that provides stability and generates functionality for the sensing of L-Cysteine (L-Cys). Being one of the important amino acid, L-cys has shown a fundamental role in living species due to the existence of sulfhydryl bonding which further affect the process of protein synthesis in living system. Therefore sensing of L-cys by using CuO@DOP NPs deserves higher consideration. Further, morphological and size parameters have been analyzed by using FESEM and HRTEM techniques. Surface interaction and coating of dopamine over CuO NPs has been examined through FTIR and TGA analysis. The non-toxicity and bio-compatibility of CuO@DOP NPs has been evaluated against L929 cell lines and on bacterial species. A computational study using DFT has been performed to check the possible mechanism of interaction between the CuO@DOP NPs and L-Cys. The higher value of detection limit (35 nM) has further shown its potential scope in sensing L-cys. The interference studies in presence of other amino acids have also taken into consideration for checking the selectivity and sensitivity of designed sensor. The applicability of prepared sensor has further been tested for real human blood serum and urine samples for detecting the presence of L-Cys. The as developed sensor of CuO@DOP NPs has provided rapid and selective sensing ability towards L-Cys over a wide range of concentration and bio-compatibility towards living entity. NOVELTY STATEMENT: Herein, the application of unique chemical and photo-luminescent properties of CuO NPs have been chosen for the fabrication of new type of fluorophore to complement conventional types of biosensor for L-Cys. The synthesis of CuO based biosensor has been achieved via an economically viable microwave assisted method. The article has not been published in any language anywhere and that it is not under simultaneous consideration by another journal. The current work is novel.

CRISPR/Cas9 engineering of albino cystinuria Type A mice.

Cystinuria Type A is a relatively common genetic kidney disease occurring in 1 in 7,000 people worldwide that results from mutation of the cystine transporter rBAT encoded by Slc3a1. We used CRISPR/Cas9 technology to engineer cystinuria Type A mice via genome editing of the C57BL/6NHsd background. These mice are an improvement on currently available models as they are on a coisogenic genetic background and have a single defined mutation. In order to use albinism to track Cas9 activity, we co-injected gRNAs targeting Slc3a1 and tyrosinase (Tyr) with Cas9 expressing plasmid DNA into mouse embryos. Two different Slc3a1 mutational alleles were derived, with homozygous mice of both demonstrating elevated urinary cystine levels, cystine crystals, and bladder stones. We used whole genome sequencing to evaluate for potential off-target editing. No off-target indels were observed for the top 10 predicted off-targets for Slc3a1 or Tyr. Therefore, we used CRISPR/Cas9 to generate coisogenic albino cystinuria Type A mice that could be used for in vivo imaging, further study, or developing new treatments of cystinuria.

Higher Levels of Low Molecular Weight Sulfur Compounds and Homocysteine Thiolactone in the Urine of Autistic Children.

In this study, the levels of concentration of homocysteine thiolactone (HTL), cysteine (Cys), and cysteinylglycine (CysGly) in the urine of autistic and non-autistic children were investigated and compared. HTL has never been analyzed in autistic children. The levels of low molecular weight sulfur compounds in the urine of both groups were determined by validated methods based on high-performance liquid chromatography with spectrofluorometric and diode-array detectors. The statistical data show a significant difference between the examined groups. Children with autism were characterized by a significantly higher level of HTL (p = 5.86 × 10-8), Cys (p = 1.49 × 10-10) and CysGly (p = 1.06 × 10-8) in urine compared with the control group. A difference in the p-value of <0.05 is statistically significant. Higher levels of HTL, Cys, and CysGly in the urine of 41 children with autism, aged 3 to 17, were observed. The obtained results may indicate disturbances in the metabolism of methionine, Cys, and glutathione in some autistic patients. These preliminary results suggest that further research with more rigorous designs and a large number of subjects is needed.

Synthesis and characterization of maltol capped silver nanoparticles and their potential application as an antimicrobial agent and colorimetric sensor for cysteine.

Maltol capped silver nanoparticles (McAgNPs) were synthesized using maltol (3-hydroxy-2-methyl-4-pyrone) as reducing and capping agent. McAgNPs were characterized by Visible and FTIR (Fourier transform infrared) spectroscopy, dynamic light scattering (DLS), and atomic force microscopy (AFM). Bright yellow color McAgNPs showed surface plasmon resonance (SPR) band at 436 nm, spherical shape and the average size between 35 to 50 nm. McAgNPs revealed higher stability against varying storage time, temperature, pH and salt concentrations. McAgNPs were successfully utilized for the selective and highly sensitive colorimetric detection of cysteine (Cys). Addition of Cys in a solution of McAgNPs, resulted a rapid change in color from yellow to orange because of the formation of nanoaggregates as confirmed by Visible/FTIR spectroscopy, DLS, and AFM studies. The estimated limit of detection (0.043 µM) was found to be more sensitive than previously reported other optical methods. The practical applicability of probe was also established by spiking the known concentrations of Cys in biological (blood plasma and urine) and environmental (tap and lake water) samples with significant recovery rates (92-104.6%). Despite being nontoxic to various tested cell lines, McAgNPs demonstrated potent antimicrobial, antibiofilm, and biofilm eradicating activities, thus potentially valuable in diagnostics and/or the synthesis of other nanocomposite material for broader applications.

A novel fluorescent probe for rapidly detection cysteine in cystinuria urine, living cancer/normal cells and BALB/c nude mice.

Cysteine (Cys), an important organic small molecule containing sulfhydryl groups, plays paramount functions in human pathologies and physiologies. The detection of Cys in living vivo is essential for studying its roles. Here, we designed and synthesized a novel red-emission fluorescent probe AXPI-Cys with highly sensitivity (LOD = 48.9 ±â€¯0.23 nM), rapidly response (<7 min) and colorimetric for detection cysteine. More importantly, the AXPI-Cys was determined Cys in real cystinuria urine samples for the first time with the satisfactory results (92%-99.96%) and employed for specifically location of endogenous/exogenous Cys in living cancer/normal cells and almost non-toxic, that is very valuable for diagnosis of cystinuria and observation of the distribution of Cys in normal cells. Notably, the AXPI-Cys was applied to imaging Cys in BALB/c nude mice with good biocompatibility and desirable tissue-penetration depth. Owing to the superior capability of AXPI-Cys, it provided a desired method to detect Cys in urine samples and cells, and exhibited munificent potential usage in biosystems and imaging studies in vivo.

An LC-MS/MS Method to Measure S-Methyl-l-Cysteine and S-Methyl-l-Cysteine Sulfoxide in Human Specimens Using Isotope Labelled Internal Standards.

This is the first report describing an analytical method for quantitative analysis of two naturally occurring sulphur compounds, S-methyl-l-cysteine (SMC) and S-methyl-l-cysteine sulfoxide (SMCSO), in human body fluids using isotope-labelled internal standards and liquid chromatography-mass spectrometry (LC-MS)/MS techniques. This method was validated according to the guideline of the Royal Society of Chemistry Analytical Methods Committee. It offers significant advantages including simple and fast preparation of human biological samples. The limits of detection of SMC were 0.08 µM for urine and 0.04 µM for plasma. The limits of detection of SMCSO were 0.03 µM for urine and 0.02 µM for plasma. The calibration curves of all matrices showed linearity with correlation coefficients r2 > 0.9987. The intra and inter day precisions in three levels of known concentrations were >10% and >20%, respectively. The quantification accuracy was 98.28 ± 5.66%. The proposed method would be beneficial for the rapid and accurate determination of the SMC and SMCSO in human plasma and urine samples using by isotope labelled internal standards.

Label-free gold nanorods sensor array for colorimetric detection and discrimination of biothiols in human urine samples.

Biothiols play important roles in regulating redox balance in biological systems, but their discrimination is challengeable. In this work, a colorimetric nanosensing array for biothiols was established, which was composed of gold nanorods (AuNRs) and metal ions (Hg2+, Pb2+, Cu2+, Ag+). By employing label-free AuNRs as the colorimetric probe, and the color and spectral changes of AuNRs as the output signal, principal component analysis (PCA) was applied to processing the signal and generating a clustering map. Due to the different binding affinity between biothiols and metal ions, AuNRs exhibited a unique pattern to form a fingerprint-like colorimetric array, which was able to discriminate five biothiols by the naked eyes. This strategy combines PCA and sensor array to achieve rapid and accurate discrimination and detection of biothiols. In addition, the method shows the great potential in analysis of biothiols in human urine samples.

Silver-ion-mediated Mg2+-dependent DNAzyme activity for amplified fluorescence detection of cysteine.

In this paper, by taking advantage of the fact that silver ions could mediate the Mg2+-dependent DNAzyme (Mgzyme) activity, we for the first time developed a turn-on fluorescent biosensor for amplified cysteine (Cys) detection. Because Mgzyme can interact with the silver ion and form cytosine-Ag+-cytosine (C-Ag+-C) base pairs, the conformation of its catalytic core was changed. As a result, the catalytic activity of Mgzyme was suppressed and the Mgzyme-Ag+ complex could not initiate the cleavage reaction. Therefore, the background fluorescence of the biosensor was very low. In the presence of Cys, Cys can bind tightly to the silver ion and disrupt the C-Ag+-C base pairs in the Mgzyme-Ag+ complex, leading to the restoration of Mgzyme activity. The activated Mgzyme could hybridize with the MB substrate and undergo many cleavage cycles, resulting in a significant increase of fluorescence intensity. This designed strategy provided amplified fluorescence detection of cysteine, with a detection limit of 2 nM. Moreover, the strong binding between Cys and Ag+ ensured that the biosensor had a desirable selectivity for Cys. This sensing system was also used to detect Cys in human urine samples and displayed satisfying results.

On-off-on fluorescent carbon dots from waste tea: Their properties, antioxidant and selective detection of CrO42-, Fe3+, ascorbic acid and L-cysteine in real samples.

In this work, we reported an economical plant-based hydrothermal method for one-pot green synthesis of water-soluble carbon dots (Tea-CDs) by using waste tea extract as a carbon source. The synthesized Tea-CDs were characterized by UV-visible, fluorescence, FT-IR, TEM, XPS and XRD. The Tea-CDs were found to remove hydroxyl and superoxide anion radical in vitro. In addition, the Tea-CDs exhibited bright blue fluorescence under UV-light (λex = 365 nm), and the fluorescence could be effectively quenched by CrO42- and Fe3+ ions. Meanwhile, the fluorescence of Tea-CDs-CrO42- and Tea-CDs-Fe3+ systems could be again easily recovered by ascorbic acid (AA) and L-cysteine (L-Cys). As an on-off-on fluorescent nano-sensor of the Tea-CDs, the sensitive detection of CrO42-, Fe3+, AA and L-Cys were all performed, showing that the good linear relationships between fluorescence intensity of Tea-CDs and concentration of all testing samples. Finally, the sensors successfully detected CrO42-, Fe3+, AA and L-Cys in commercially available real samples with satisfactory recovery ranges. The prepared sensors offer distinct advantages including low cost, simple handling, good sensitivity and high selectivity.

Metabolomic Profiling of Infants With Recurrent Wheezing After Bronchiolitis.

BACKGROUND: Bronchiolitis is associated with a greater risk of developing recurrent wheezing, but with currently available tools, it is impossible to know which infants with bronchiolitis will develop this condition. This preliminary prospective study aimed to assess whether urine metabolomic analysis can be used to identify children with bronchiolitis who are at risk of developing recurrent wheezing. METHODS: Fifty-two infants <1 year old treated in the emergency department at University Hospital of Padova for acute bronchiolitis were enrolled (77% tested positive for respiratory syncytial virus [RSV]). Follow-up visits were conducted for 2 years after the episode of bronchiolitis. Untargeted metabolomic analyses based on mass spectrometry were performed on urine samples collected from infants with acute bronchiolitis. Data modeling was based on univariate and multivariate data analyses. RESULTS: We distinguished children with and those without postbronchiolitis recurrent wheeze, defined as ≥3 episodes of physician-diagnosed wheezing. Pathway overrepresentation analysis pointed to a major involvement of the citric acid cycle (P < .001) and some amino acids (lysine, cysteine, and methionine; P ≤ .015) in differentiating between these 2 groups of children. CONCLUSION: This is the first study showing that metabolomic profiling of urine specimens from infants with bronchiolitis can be used to identify children at increased risk of developing recurrent wheezing.

An On-Off-On Fluorescence Probe Based on Coumarin for Cu2+, Cysteine, and Histidine Detections.

A simple coumarin-based Schiff base (probe L) was successfully developed. It showed strong green fluorescence emission at 527 nm with a 70.3% of fluorescence quantum efficiency (ΦF). However, after the addition of common metal ions, probe L can only combine with Cu2+ ions and displayed significant fluorescence quenching of > 96.2% (ΦF = 2.7%) due to the paramagnetic quenching action from Cu2+. Conversely, by the coordination action of cysteine (Cys) and histidine (His), the quenching fluorescence of the complex (L-Cu2+) between probe L and Cu2+ ions was recovered mostly because the Cys and His can usurp Cu2+ of L-Cu2+ and led to the liberation of probe L. Based on the fluorescence changes of probe L with the actions of Cu2+, Cys, and His, an on-off-on reversible fluorescence probe for sensitive and specific monitoring Cu2+, Cys, and His has been prepared. More importantly, the probe L and L-Cu2+ ensemble can be used, respectively, to test Cu2+ and Cys/His in live cells and human urine samples with great reliability.

Urine concentrations changes of cysteinyl leukotrienes in non-obese children with obstructive sleep apnea undergoing adenotonsillectomy.

OBJECTIVE: The main objective of the study was to compare preoperative to postoperative levels of urine-Cysteinyl leukotrienes (uCysLT) in children undergoing adenotonsillectomy (AT) for obstructive sleep apnea (OSA) in order to investigate whether exaggerated leukotriene activity is the cause or consequence of OSA. METHODS AND MATERIALS: A prospective study was conducted on non-obese children (4-10 years old) referred for overnight PSG. Children with moderate/severe OSA treated with AT were included. A second PSG study performed 2 months postoperatively to confirm OSA resolution, and those with residual OSA were excluded. Morning urine specimens after both PSG studies were obtained and pre-operative uCysLT levels were compared to postoperative levels. RESULTS: 27 children fulfilled the criteria and underwent a post-operative PSG study with three exclusions for residual OSA (postop-AHI>2), so the study group consisted of 24 children (mean age: 5.7 ±â€¯2.1 years). Mean preoperative and postoperative AHI was 10.96 ±â€¯5.93 and 1.44 ±â€¯0.56 respectively. Mean preop-uCysLT were 21.14 ±â€¯4.65, while after AT they significantly reduced to 12.62 ±â€¯2.67 (P < 0.01). CONCLUSION: uCysLT levels are significantly reduced after AT in non-obese children with moderate/severe OSA, suggesting that exaggerated leukotriene activity is mainly a consequence of OSA.

Red emission nitrogen, boron, sulfur co-doped carbon dots for "on-off-on" fluorescent mode detection of Ag+ ions and l-cysteine in complex biological fluids and living cells.

Herein, a simple and efficient fluorescent assay for Ag+ ions and l-cysteine (L-Cys) in complex biological fluids and living cells was first developed based on the fluorescent "on-off-on" mode of red emission nitrogen, boron, sulfur co-doped carbon dots (NBS-CDs). Red emission NBS-CDs were prepared via one-step hydrothermal synthesis by using 3-aminobenzeneboronic acid and 2,5-diaminobenzenesulfonic acid as precursors. Such NBS-CDs exhibited excellent optical properties and relatively high absolute fluorescent quantum yield compared with some reported NBS-CDs. Due to the strong quenching ability of Ag+ ions on the fluorescence of NBS-CDs, red emission NBS-CDs were used for the determination of Ag+ ions with high sensitivity and excellent selectivity. The fluorescence of NBS-CDs was recovered after the interaction between Ag+ ions and L-Cys, which realized the specific determination of L-Cys in human urine samples and human plasma samples. The established NBS-CDs-based fluorescent "on-off-on" sensor offered a relatively low detection limits of 0.35 µM for Ag+ ions and 0.045 µM for L-Cys based on three times signal-to-noise criteria. Notably, this strategy was applied for the visual detections of Ag+ ions and L-Cys in living human cancer cells (HeLa cells and MCF-7 cells). This method is simple, high sensitive, and excellent selectivity, which provided a new insight on the potential applications of NBS-CDs to develop the biosensor in clinical diagnosis and other biologically related areas.

Cysteinyl-leukotriene pathway as a new therapeutic target for the treatment of atherosclerosis related to obstructive sleep apnea syndrome.

AIMS: Obstructive sleep apnea (OSA) characterized by nocturnal intermittent hypoxia (IH) is associated with atherosclerosis and cysteinyl-leukotrienes (CysLT) pathway activation. We aimed to identify the determinants of CysLT pathway activation and the role of CysLT in OSA-related atherosclerosis. METHODS AND RESULTS: Determinants of the urinary excretion of LTE4 (U-LTE4) including history of cardiovascular events, polysomnographic and biological parameters were studied in a cohort of 170 OSA patients and 29 controls, and in a subgroup of OSA patients free of cardiovascular event (n = 136). Mechanisms linking IH, the CysLT pathway and atherogenesis were investigated in Apolipoprotein E deficient (ApoE-/-) mice exposed to 8-week IH. In the whole cohort, U-LTE4 was independently influenced by age, minimal oxygen saturation, and a history of cardiovascular events, and correlated significantly with intima-media thickness. In the subgroup of OSA patients free of cardiovascular event, increased U-LTE4 was increased compared to controls and independently related to hypoxia severity and traditional risk factors aggregated in the 10-year cardiovascular risk score of European Society of Cardiology. In IH mice, atherosclerosis lesion size and mRNA levels of 5-lipoxygenase, 5-lipoxygenase activating protein (FLAP) and CysLT1 receptor were significantly increased. This transcriptional activation was associated with the binding of HIF-1 to the FLAP promoter and was strongly associated with atherosclerosis lesion size. CysLT1 receptor antagonism (montelukast) significantly reduced atherosclerosis progression in IH mice. CONCLUSIONS: IH-related CysLT pathway activation contributes to OSA-induced atherogenesis. In the era of personalized medicine, U-LTE4 may be a useful biomarker to identify OSA patients for whom CysLT1 blockade could represent a new therapeutic avenue for reducing cardiovascular risk.

Development of a rapid UPLC-MS/MS determination of urine sulfocysteine for diagnosis of sulfocysteinuria and molybdenum co-factor deficiencies.

AIM: Molybdenum co-factor deficiencies and isolated sulfite oxidase deficiency are rare autosomal recessively inherited diseases characterized by severe psychomotor impairment, intractable seizures, dislocated lens and dysmorphic facial features. The biochemical diagnosis of these diseases requires the determination of urine sulfocysteine. MATERIALS & METHODS: Urine sulfocysteine was quantified by an ultra-high performance liquid chromatography-MS/MS assay. The method was validated for linearity, accuracy, precision, recovery and stability. RESULTS & CONCLUSION: Total imprecision of accuracy was less than 6%. Intra-assay and inter-assay precisions were less than 5%. The recovery was higher than 98%. The method is inexpensive, fast, accurate and has been successfully used for identifying five molybdenum co-factor deficient and six sulfite oxidase deficient patients since deployed.

Molecular Vise Approach to Create Metal-Binding Sites in MOFs and Detection of Biomarkers.

We report a new approach to create metal-binding site in a series of metal-organic frameworks (MOFs), where tetratopic carboxylate linker, 4',4'',4''',4''''-methanetetrayltetrabiphenyl-4-carboxylic acid, is partially replaced by a tritopic carboxylate linker, tris(4-carboxybiphenyl)amine, in combination with monotopic linkers, formic acid, trifluoroacetic acid, benzoic acid, isonicotinic acid, 4-chlorobenzoic acid, and 4-nitrobenzoic acid, respectively. The distance between these paired-up linkers can be precisely controlled, ranging from 5.4 to 10.8 Å, where a variety of metals, Mg2+ , Al3+ , Cr3+ , Mn2+ , Fe3+ , Co2+ , Ni2+ , Cu2+ , Zn2+ , Ag+ , Cd2+ and Pb2+ , can be placed in. The distribution of these metal-binding sites across a single crystal is visualized by 3D tomography of laser scanning confocal microscopy with a resolution of 10 nm. The binding affinity between the metal and its binding-site in MOF can be varied in a large range (observed binding constants, Kobs from 1.56×102 to 1.70×104  L mol-1 ), in aqueous solution. The fluorescence of these crystals can be used to detect biomarkers, such as cysteine, homocysteine and glutathione, with ultrahigh sensitivity and without the interference of urine, through the dissociation of metal ions from their binding sites.