An Electrochemiluminescent Sensor Based on Glycosyl Imprinting and Aptamer for the Detection of Cancer-Related Extracellular Vesicles.

Cancer-related extracellular vesicles (EVs) are considered important biomarkers for cancer diagnosis because they can convey a large amount of information about tumor cells. In order to detect cancer-related EVs efficiently, an electrochemiluminescence (ECL) sensor for the specific identification and highly sensitive detection of EVs in the plasma of cancer patients was constructed based on dual recognitions by glycosyl-imprinted polymer (GIP) and aptamer. The characteristic glycosyl Neu5Ac-α-(2,6)-Gal-ß-(1-4)-GlcNAc trisaccharide on the surface of EVs was used as a template molecule and 3-aminophenylboronic acid as a functional monomer to form a glycosyl-imprinted polymer by electropolymerization. After glycosyl elution, the imprinted film specifically recognized and adsorbed the EVs in the sample, and then the CD63 aptamer-bipyridine ruthenium (Aptamer-Ru(bpy)) was added to combine with the CD63 glycoprotein on the extracellular vesicle's surface, thus providing secondary recognition of the EVs. Finally, the EVs were quantitatively detected according to the ECL signal produced by the labeled bipyridine ruthenium. When more EVs were captured by the imprinted film, more probes were obtained after incubation, and the ECL signal was stronger. Under the optimized conditions, the ECL signal showed a good linear relationship with the concentration of EVs in the range of 9.5 × 102 to 9.5 × 107 particles/mL, and the limit of detection was 641 particles/mL. The GIP sensor can discriminate between the EV contents of cancer patients and healthy controls with high accuracy. Because of its affordability, high sensitivity, and ease of use, it is anticipated to be employed for cancer early detection and diagnosis.

A molecularly imprinted photoelectrochemical sensor based on an rGO/MoSSe heterojunction for the detection of chlortetracycline.

A reduced graphene oxide/molybdenum selenosulfide (rGO/MoSSe) heterojunction was synthesized, and a molecularly imprinted photoelectrochemical sensor for the detection of chlortetracycline was prepared. MoSSe was grown in situ on rGO by a hydrothermal method to form an rGO/MoSSe heterojunction, which acts as the sensitive film of the sensor. Since rGO can promote electron transfer and effectively inhibit electron-hole recombination, it effectively reduces the recombination probability of electrons and holes and improves the photoelectric efficiency, thus enhancing the detection sensitivity of the PEC sensor. The rGO/MoSSe was immobilized on an FTO electrode, and molecularly imprinted polymers (MIPs) were prepared by electropolymerization on the rGO/MoSSe-modified FTO electrode with chlortetracycline as the template molecule and o-phenylenediamine as the functional monomer, so as to construct a molecularly imprinted photoelectrochemical (MIP-PEC) sensor. The determination of chlortetracycline was realized by the strategy of a "gate-controlled effect", and the detection range of the chlortetracycline concentration was 5.0 × 10-13-5 × 10-9 mol L-1 with a detection limit of 1.57 × 10-13 mol L-1. The sensor has been applied to the determination of chlortetracycline in animal-derived food samples.

Electrochemical chiral sensor for levofloxacin detection base on Cu/Fe-BTC amplification.

The key to developing sensors for chiral drug determination is to exclude interference from enantiomers. In this study, metal-organic frameworks (MOFs) and molecularly imprinted polymer (MIP) were introduced to prepare a chiral sensor for levofloxacin detection. The MIP was electropolymerised on the surface of the Cu/Fe-benzene-1,3,5-tricarboxylate MOF (Cu/Fe-BTC)-modified Au electrode using levofloxacin as a template molecule. After eluting the levofloxacin, a chiral sensor with recognition sites for levofloxacin was obtained. With this site as a switch, a novel method for detecting levofloxacin was established. Because of the enhanced recognition effect, the sensor can effectively exclude the enantiomeric interference of d-ofloxacin. Moreover, Cu/Fe-BTC can effectively amplify the current response signal and improve the sensitivity of the sensor. The linear range of the sensor was 5 to 4000 × 10-11 mol L-1, and the detection limit was 2.07 × 10-11 mol L-1. When applied to detecting levofloxacin in actual samples, the sensor showed a 92.7-109.8% recovery.

Application of Computer-Aided Design and Individualized Templates for Bilateral Zygomaticomaxillary Complex Fractures.

PURPOSE: Reduction of the bilateral zygomaticomaxillary complex (ZMC) fracture with individualized templates based on computer- aided surgical simulation system. To evaluate the practicality and accuracy of this approach in the treatment of bilateral ZMC fracture. METHODS: Sixteen patients with bilateral ZMC fractures were collected to create a study model. The authors reconstruct the ZMC on one side via the three-dimensional (3D) model, and then mirrored to the opposite side. Multiple individualized templates were made based on the 3D model, and used as intraoperative guidance to reduce fractures. After surgery, the facial symmetry and the position of zygoma were observed. The mouth opening, pupil level, and sensation of infraorbital nerve were evaluated. Some mark points on zygoma were measured and the postoperative horizontal asymmetry rate (H) was calculated. Besides, orbital height and width were measured. RESULTS: For all patients, the position of bilateral ZMC was basically restored. The patients with restriction of mouth opening all recovered to normal. The H values were less than 3.0% at all mark points. There was almost no difference in bilateral orbital width and height. Meanwhile, there was no significant difference between the preoperative measurements of the ideal virtual 3D model and the postoperative measurements of patients. CONCLUSIONS: The study proves that application of computer-aided design and individualized templates can accurately guide the reduction operation of ZMC fracture, restore the ideal shape of ZMC, and obtain good facial symmetry.

A versatile and ultrasensitive molecularly imprinted electrochemiluminescence sensor with HRP-encapsulated liposome labeled by light-triggered click reaction for pesticide residues.

A novel approach for trace detection of fipronil with a molecularly imprinted electrochemiluminescence sensor (MIECLS) is proposed. The sensitivity is significantly improved via signal amplification of the enzymatic reaction of horseradish peroxidase (HRP) released from encapsulated liposomes which linked onto the template molecules after rebinding. The molecularly imprinted polymer membrane was prepared through the electropolymerization of monomers with fipronil as a template. After the elution of the template molecules, the analyte fipronil was reabsorbed into the cavities. HRP-encapsulated liposomes were linked to the target molecules by light-triggered click reaction. The higher the concentration of the target was, the more HRP-encapsulated liposomes were present on the molecularly imprinted polymer (MIP) sensor. Then, HRP was liberated from liposomes, and the catalytic degradation of hydrogen peroxide (H2O2) by HRP occurs, which changed the electrochemiluminescence intensity of luminol significantly. The change of the ∆ECL intensity was linearly proportional to the logarithm of the fipronil concentration ranging from 1.00 × 10-14 to 1.00 × 10-9 mol/L, and the detection limit was 7.77 × 10-16 mol/L. The recoveries obtained ranged from 95.7 to 105.8% with RSD < 5%. The sensitivity of the detection was significantly improved, and the analysis process was simplified in that the incubation step required in the conventional method was avoided. The sensor proposed provides a feasible platform for ultra-trace amount determination.

Magnetic stir cake sorptive extraction of trace tetracycline antibiotics in food samples: preparation of metal-organic framework-embedded polyHIPE monolithic composites, validation and application.

In this work, a novel Fe3O4@Cu3(btc)2-embedded polymerized high internal phase emulsion (Fe3O4@HKUST-1-polyHIPE) monolithic cake was synthesized, characterized and used as an adsorbent in the magnetic stir cake sorptive extraction (MSCSE) and determination of tetracycline antibiotics (TCs) in food samples by a combination of with high-performance liquid chromatography-fluorescence detection (HPLC-FLD). The prepared Fe3O4@HKUST-1-polyHIPE monolithic composites displayed a strong extraction ability and high column capacity due to enhanced interactions such as π-π interactions, hydrogen bonding, and electrostatic interactions. The extraction and desorption conditions were evaluated, and the calibration curves of four spiked TCs were linear (R2 ≥ 0.9991) in the range from 20 to 800 ng mL-1 for milk and egg samples, and 20 to 800 ng g-1 for chicken muscle and kidney samples. The limits of detection and the limits of quantification of the four TCs by using the proposed MSCSE-HPLC-FLD method were in the range of 1.9-4.6 and 5.5-13.9 ng mL-1 for milk and egg samples, and 1.8-3.7 and 5.3-13.0 ng g-1 for chicken muscle and kidney samples, respectively. The recoveries of the target TCs from spiked food samples were in the range from 86.6 to 110.7% with relative standard deviations lower than 7.0%. The proposed method was successfully applied for the determination of these four TCs in milk, egg, chicken muscle, and kidney samples.

Sensitive electrochemical detection of sodium azide based on the electrocatalytic activity of the pasting liquid of a carbon paste electrode.

Sodium azide (NaN3) is highly toxic and widely used in, for example, automobile airbags and biochemical laboratories. The electrochemical detection of sodium azide on commonly used electrodes is challenging due to sluggish electron transfer, but it has been achieved using a boron-doped diamond thin-film electrode and a highly oriented pyrolytic graphite electrode. Utilizing the electrocatalytic activity of the pasting liquid of a carbon paste electrode, we developed an effective method for the electrochemical detection of sodium azide in which silicone oil was employed as the pasting liquid of the carbon paste electrode. This simple and cheap silicone-oil-based carbon paste electrode exhibited comparable sensitivity to the boron-doped diamond thin-film electrode and highly oriented pyrolytic graphite electrode. The limit of detection for sodium azide at the silicone-oil-based carbon paste electrode was found to be 0.1 µM. Recoveries from diluted human serum samples were between 97.2 and 101.3%. Graphical abstract ᅟ.

Chiral determination of cinchonine using an electrochemiluminescent sensor with molecularly imprinted membrane on the surfaces of magnetic particles.

A novel molecular imprinting electrochemiluminescence sensor for detecting chiral cinchonine molecules was developed with a molecularly imprinted polymer membrane on the surfaces of magnetic microspheres. Fe3 O4 @Au nanoparticles modified with 6-mercapto-beta-cyclodextrin were used as a carrier, cinchonine as a template molecule, methacrylic acid as a functional monomer and N,N'-methylenebisacrylamide as a cross-linking agent. Cinchonine was specifically recognized by the 6-mercapto-beta-cyclodextrin functional molecularly imprinted polymer and detected based on enhancement of the electrochemiluminescence intensity caused by the reaction of tertiary amino structures of cinchonine molecules with Ru(bpy)32+ . Cinchonine concentrations of 1 × 10-10 to 4 × 10-7  mol/L showed a good linear relationship with changes of the electrochemiluminescence intensity, and the detection limit of the sensor was 3.13 × 10-11  mol/L. The sensor has high sensitivity and selectivity, and is easy to renew. It was designed for detecting serum samples, with recovery rates of 98.2% to 107.6%.

A molecularly imprinted electrochemiluminescence sensor based on the mimetic enzyme catalytic effect for ultra-trace Ni2+ determination.

A novel molecularly imprinted polymer (MIP) electrochemiluminescence (MIP-ECL) sensor was developed for the highly sensitive and selective determination of ultra-trace levels of Ni2+. The complex Ni2+-dimethylglyoxime (Ni-DMG) was chosen as the template molecule to construct the MIP and then acted as a mimetic enzyme to catalyse the oxidisation of luminol to enhance the ECL signal. When the imprinted cavities were occupied by Ni-DMG in the rebinding process, the ECL intensities produced by the luminol-H2O2 ECL system on the MIP-modified electrode surface increased with increased concentration of the Ni-DMG complex. The highly sensitive determination of Ni2+ was achieved through a catalytic reaction. This technique could be used for the quantitative analysis of Ni2+ with concentrations from 3.0 × 10-12 mol L-1 to 6.0 × 10-9 mol L-1. The detection limit was 1.01 × 10-12 mol L-1, which is much lower than that reported previously. In addition, the allowable amounts of interference ions in the MIP-ECL sensor were higher than that in other common molecularly imprinted sensors because of its excellent recognition of 3D cavity-to-complex molecules and ligand-to-metal ions. This method was successfully used to determine Ni2+ in real samples, such as apples, carrots and grapes, and has been proven feasible for practical applications.

A molecularly imprinted sensor based on an electrochemiluminescent membrane for ultratrace doxycycline determination.

A new molecularly imprinted sensor was developed based on an electroluminescent molecularly imprinted polymer (MIP) membrane and used for doxycycline determination. The MIP was prepared by electropolymerization of pyrogallol doped with alizarin red. An electrochemiluminescence (ECL) signal was produced by the oxidation of the poly-pyrogallol polymer and reaction with alizarin red. The luminescence intensity was enhanced by doxycycline molecules which were re-adsorbed in cavities in MIP due to the energy transfer of the doxycycline oxidized intermediate to alizarin red. The changes of ECL intensities were linear with the concentrations of doxycycline in the range of 2 × 10(-10) to 5 × 10(-8) mol L(-1). The detection limit was 5.17 × 10(-11) mol L(-1). This method was utilized to determine doxycycline residuals in fish muscles with satisfactory results.

High-internal-phase-emulsion polymeric monolith coupled with liquid chromatography-electrospray tandem mass spectrometry for enrichment and sensitive detection of trace cytokinins in plant samples.

High-internal-phase-emulsion polymers (polyHIPEs) show great promise as solid-phase-extraction (SPE) materials because of the tremendous porosity and highly interconnected framework afforded by the high-internal-phase-emulsion (HIPE) technique. In this work, polyHIPE monolithic columns as novel SPE materials were prepared and applied to trace enrichment of cytokinins (CKs) from complex plant samples. The polyHIPE monoliths were synthesized via the in-situ polymerization of the continuous phase of a HIPE containing styrene (STY) and divinylbenzene (DVB) in a stainless column, and revealed highly efficient and selective enrichment ability for aromatic compounds. Under the optimized experimental conditions, a method using a monolithic polyHIPE column combined with liquid chromatography-electrospray tandem mass spectrometry (LC-MS-MS) was developed for the simultaneous extraction and sensitive determination of trans-zeatin (tZ), meta-topolin (mT), kinetin (K), and kinetin riboside (KR). The proposed method had good linearity, with correlation coefficients (R (2)) from 0.9957 to 0.9984, and low detection limits (LODs, S/N = 3) in the range 2.4-47 pg mL(-1) for the four CKs. The method was successfully applied to the determination of CKs in real plant samples, and obtained good recoveries ranging from 68.8 % to 103.0 % and relative standard deviations (RSDs) lower than 16 %.

Application of mercapto-silica polymerized high internal phase emulsions for the solid-phase extraction and preconcentration of trace lead(II).

A new class of solid-phase extraction column prepared with grafted mercapto-silica polymerized high internal phase emulsion particles was used for the preconcentration of trace lead. First, mercapto-silica polymerized high internal phase emulsion particles were synthesized by using high internal phase emulsion polymerization and carefully assembled in a polyethylene syringe column. The influences of various parameters including adsorption pH value, adsorption and desorption solvents, flow rate of the adsorption and desorption procedure were optimized, respectively, and the suitable uploading sample volumes, adsorption capacity, and reusability of solid phase extraction column were also investigated. Under the optimum conditions, Pb(2+) could be preconcentrated quantitatively over a wide pH range (2.0-5.0). In the presence of foreign ions, such as Na(+) , K(+) , Ca(2+) , Zn(2+) , Mg(2+) , Cu(2+) , Fe(2+) , Cd(2+) , Cl(-) and NO3 (-) , Pb(2+) could be recovered successfully. The prepared solid-phase extraction column performed with high stability and desirable durability, which allowed more than 100 replicate extractions without measurable changes of performance. The feasibility of the developed method was further validated by the extraction of Pb(2+) in rice samples. At three spiked levels of 40.0, 200 and 800 µg/kg, the average recoveries for Pb(2+) in rice samples ranged from 87.3 to 105.2%.

Protein synergistic action-based development and application of a molecularly imprinted chiral sensor for highly stereoselective recognition of S-fluoxetine.

In order to improve the recognition performance of MIPs sensors in chiral drug enantiomers, a novel a highly selective molecular recognition method based on protein-assisted immobilization of chiral molecular conformation was developed. S-fluoxetine (S-FLX) as the target chiral molecule, human serum albumin (HSA), which has a high affinity and strong interactions with S-FLX, was screened from 11 proteins to serve as an auxiliary recognition unit for the fixation of chiral conformation. By incorporating HSA into the preparation of molecularly imprinted polymers (MIPs), the natural chirality and high stereoselectivity of the protein were leveraged for the induction and fixation of the stereo conformation of S-FLX, refinement of internal structures of the imprinted cavities. The sensor exhibited excellent chiral recognition ability and high detection sensitivity. The changes of probe signal intensity of the MIPs/HSA sensor were positively correlated with the logarithmic concentration of S-FLX in the range of 1.0 × 10-16-1.0 × 10-11 mol L-1, where a detection limit of 6.43 × 10-17 mol L-1 was achieved (DL = 3δb/K). The selectivity of MIPs/HSA sensor in recognizing S-FLX was increased by 18.5 times and the sensitivity was increased by 2.6 times after the incorporation of HSA. The developed sensor was successfully used for the analysis of S-FLX in fluoxetine hydrochloride capsules.

Development of Levo-Lansoprazole Chiral Molecularly Imprinted Polymer Sensor Based on the Polylysine-Phenylalanine Complex Framework Conformational Separation.

The efficacies and toxicities of chiral drug enantiomers are often dissimilar, necessitating chiral recognition methods. Herein, a polylysine-phenylalanine complex framework was used to prepare molecularly imprinted polymers (MIPs) as sensors with enhanced specific recognition capabilities for levo-lansoprazole. The properties of the MIP sensor were investigated using Fourier-transform infrared spectroscopy and electrochemical methods. The optimal sensor performance was achieved by applying self-assembly times of 30.0 and 25.0 min for the complex framework and levo-lansoprazole, respectively, eight electropolymerization cycles with o-phenylenediamine as the functional monomer, an elution time of 5.0 min using an ethanol/acetic acid/H2O mixture (2/3/8, V/V/V) as the eluent, and a rebound time of 10.0 min. A linear relationship was observed between the sensor response intensity (ΔI) and logarithm of the levo-lansoprazole concentration (l-g C) in the range of 1.0 × 10-13-3.0 × 10-11 mol/L. Compared with a conventional MIP sensor, the proposed sensor showed more efficient enantiomeric recognition, with high selectivity and specificity for levo-lansoprazole. The sensor was successfully applied to levo-lansoprazole detection in enteric-coated lansoprazole tablets, thus demonstrating its suitability for practical applications.

A molecularly imprinted sensor for single-molecule detection of pesticide metabolite at the amol/L level sensitized by water-soluble luminol derivative encapsulated liposome via click reaction.

A novel luminol derivative, 4-[(1,4-dioxo-1,2,3,4-tetrahydrophthalazin-5-yl)amino]-4-oxobut-2-enoic acid (ALD) with electrochemiluminescence intensity and stability characteristics similar to luminol, but higher solubility in near neutral solution, was designed and synthesized in this study. Using this derivative, a molecular imprinted electrochemiluminescence sensor (MIECLS) was prepared for the sensitive and selective determination of 2-amino-5-mercapto-1,3,4-thiadiazole (AMT), a metabolite of bismerthiazol, thiediazole copper, thiazole zinc, and other pesticides. The ALD probes encapsulated in liposomes are immobilized on the molecularly imprinted film by light-triggered click reaction, and the concurrent release of multiple probes allows for highly sensitive detection. In the AMT concentration range of 1.00 × 10-18 - 5.00 × 10-13 mol/L, the relation between ECL response and log AMT concentration is linear. With a detection limit of 5.25 × 10-19 mol/L (about 4 - 6 molecules in 10 µL of the sample), the sensor allows for high sensitivity analysis of ultra-trace amounts of small organic compounds. In general, the ECL-based single-molecule detection technique proposed herein might be a promising alternative to fluorescence single-molecule detection.

Research and Application of Highly Selective Molecular Imprinting Technology in Chiral Separation Analysis.

Since residual chiral pollutants in the environment and toxic or ineffective chiral components in drugs can threat human health, there is an urgent need for methods to separation and analyze chiral molecules. Molecular imprinting technology (MIT) is a biomimetic technique for specific recognition of analytes with high potential for application in the field of chiral separation and analysis. However, since MIT has some disadvantages when used for chiral recognition, such as poor rigidity of imprinted materials, a single type of recognition site, and poor stereoselectivity, reducing the interference of conformationally and structurally similar substances to increase the efficiency of chiral recognition is difficult. Therefore, improving the rigidity of imprinted materials, increasing the types of imprinted cavity recognition sites, and constructing an imprinted microenvironment for highly selective chiral recognition are necessary for the accurate identification of chiral substances. In this article, the principle of chiral imprinting recognition is introduced, and various strategies that improve the selectivity of chiral imprinting, using derivative functional monomers, supramolecular compounds, chiral assembly materials, and biomolecules, are reviewed in the past 10 years.

Highly sensitive molecularly imprinted electrochemical sensor based on the double amplification by an inorganic Prussian blue catalytic polymer and the enzymatic effect of glucose oxidase.

A novel strategy to improve the sensitivity of molecularly imprinted polymer (MIP) sensors was proposed. An electrocatalytic Prussian blue (PB) film was electrochemically polymerized on an electrode surface to fabricate an MIP electrochemical sensor using oxytetracycline (OTC) as a template. The OTC determination relied on a competition reaction between OTC and glucose-oxidase-labeled OTC and the catalytic reduction of hydrogen peroxide by the modified PB film. Experimental results show that double amplification, which is based on the catalysis of inorganic PB films and the enzymatic effect of glucose oxidase, can remarkably increase the assay sensitivity. The main experimental conditions (including electrocatalysis of the PB film, pH effects, incubation and competition times, and anti-interference) were optimized. This novel MIP sensor can offer an femtomole detection limit for OTC. In addition, the feasibility of its practical applications has been demonstrated in the analysis of a series of real milk samples.

Molecularly imprinted electrochemical luminescence sensor based on signal amplification for selective determination of trace gibberellin A3.

A new molecularly imprinted electrochemical luminescence (MIP-ECL) sensor was developed for Gibberellin A3 (GA3) determination. This sensor is based on competitive binding between the GA3 and the Rhodamine B (RhB)-labeled GA3 (RhB-GA3) to the MIP film. After the competitive binding, the residual RhB-GA3 on the MIP was electro-oxidized to produce RhB oxide, which could greatly amplify the weak electrochemiluminescence (ECL) signal of luminol. The ECL intensity decreased when the RhB-GA3 was replaced by GA3 molecules in the samples. Accordingly, GA3 was determined in the concentration range from 1 × 10(-11) to 3 × 10(-9) mol/L with a detection limit of 3.45 × 10(-12) mol/L. The sensor shows high sensitivity and selectivity, wide response range, good accuracy, and fast response. Beer samples were assayed by using the sensors, and the recoveries ranging from 96.0% to 103.2% were obtained.

Liposomes: Preparation, Characteristics, and Application Strategies in Analytical Chemistry.

In recent years, the application of liposomes in the field of analytical chemistry has attracted more and more attention. In this paper, the preparation and characterization of liposomes are briefly summarized, and the research and application of liposomes in organic and biological substances analysis are mainly reviewed, including immobilized liposome chromatography, liposome capillary electrophoresis, liposome application in electrochemical analysis, and optical analysis. The future developments of liposomes in the assay are also prospected.

Zoledronic acid inhibits osteoclastogenesis and bone resorptive function by suppressing RANKL­mediated NF­κB and JNK and their downstream signalling pathways.

Targeting excessive osteoclast differentiation and activity is considered a valid therapeutic approach for osteoporosis. Zoledronic acid (ZOL) plays a pivotal role in regulating bone mineral density. However, the exact molecular mechanisms responsible for the inhibitory effects of ZOL on receptor activator of nuclear factor (NF)­κB ligand (RANKL)­induced osteoclast formation are not entirely clear. The present study aimed to investigate the role of ZOL in osteoclast differentiation and function, and to determine whether NF­κB and mitogen­activated protein kinase, and their downstream signalling pathways, are involved in this process. RAW264.7 cells were cultured with RANKL for differentiation into osteoclasts, in either the presence or absence of ZOL. Osteoclast formation was observed by tartrate­resistant acid phosphatase staining and bone resorption pit assays using dentine slices. The expression of osteoclast­specific molecules was analysed using reverse transcription­quantitative polymerase chain reaction and western blotting assays to deduce the molecular mechanisms underlying the role of ZOL in osteoclastogenesis. The results showed that ZOL significantly attenuated osteoclastogenesis and bone resorptive capacity in vitro. ZOL also suppressed the activation of NF­κB and the phosphorylation of c­Jun N­terminal kinase. Furthermore, it inhibited the expression of the downstream factors c­Jun, c­Fos and nuclear factor of activated T cells c1, thereby decreasing the expression of dendritic cell­specific transmembrane protein and other osteoclast­specific markers. In conclusion, ZOL may have therapeutic potential for osteoporosis.