Botulinum Neurotoxin C Dual Detection through Immunological Recognition and Endopeptidase Activity Using Porous Silicon Interferometers.

Botulinum neurotoxins (BoNTs) are the most potent toxins known in nature produced by Clostridium botulinum strains, which can cause life-threatening diseases in both humans and animals. The latter is of serious environmental and economic concern, resulting in high mortality, production losses, and rejection of contaminated animal feed. The available in vivo mouse assay is inadequate for real-time and on-site assessment of outbreaks. Herein, we present a reflective-based approach for the detection of BoNT/C while estimating its activity. Two adjacent porous Si Fabry-Pérot interferometers are simultaneously utilized to quantify minute BoNT/C concentrations by a competitive immunoassay and to assess their endopeptidase activity. The reflectivity signals of each interferometer are amplified by biochemical reaction products infiltration into the scaffold or by peptide fragments detachment from the nanostructure. The optical assay is highly sensitive in compliance with the in vivo approach by presenting a detection limit of 4.24 pg mL-1. The specificity and selectivity of the designed platform are cross-validated against BoNT/B and BoNT/D, also relevant to animal health. Finally, the analytical performances of both interferometers for real-life scenarios are confirmed using actual toxins while depicting excellent compliance to complex media analysis. Overall, the presented sensing scheme offers an efficient, rapid, and label-free approach for potential biodiagnostic elucidation of botulism outbreaks.

Real-time detection of copper contaminants in environmental water using porous silicon Fabry-Pérot interferometers.

Water sources are vulnerable to intentional and inadvertent human pollution with thousands of synthetic and geogenic trace contaminants, posing long-term effects on the aquatic ecosystem and human health. Thus, early and rapid detection of water pollutants followed by corrective and preventive actions can lead to the reduction of the overall polluting impact to safeguard public health. This study presents a generic sensing assay for the label-free detection of copper contaminants in environmental water samples using multilayered polyethylenimine (PEI) functionalized porous silicon Fabry-Pérot interferometers. The selective chelating activity of PEI thin-films was monitored in real-time by reflective interferometric Fourier transform spectroscopy (RIFTS) while assessing the improved optical responses. The optimized scaffold of two sequential PEI layers depicted a linear working range between 0.2 and 2 ppm while presenting a detection limit of 0.053 ppm (53 ppb). The specificity of the developed platform was cross-validated against various metallic pollutants and cations commonly found in water bodies (i.e., Cd2+, Pb2+, Cr3+, Fe3+, Mg2+, Ca2+, Zn2+, K+ and Al3+). Finally, as a proof of concept, the analytical performance of the porous interferometers for real-life scenarios was demonstrated in three water samples (tap, ground and irrigation), presenting sufficient adaptability to complex matrix analysis with recovery values of 85-106%. Overall, the developed sensing concept offers an efficient, rapid and label-free methodology that can be potentially adopted for routine on-site detection using a simple and portable device.

Development and Characterization of Integrated Nano-Sensors for Organic Residues and pH Field Detection.

Meeting global water quality standards is a real challenge to ensure that food crops and livestock are fit for consumption, as well as for human health in general. A major hurdle affecting the detection of pollutants in water reservoirs is the lapse of time between the sampling moment and the availability of the laboratory-based results. Here, we report the preparation, characterization, and performance assessment of an innovative sensor for the rapid detection of organic residue levels and pH in water samples. The sensor is based on carbonaceous nanomaterials (CNMs) coated with an intrinsically conductive polymer, polyaniline (PANI). Inverse emulsion polymerizations of aniline in the presence of carbon nanotubes (CNTs) or graphene were prepared and confirmed by thermogravimetric analysis and high-resolution scanning electron microscopy. Aminophenol and phenol were used as proxies for organic residue detection. The PANI/CNM nanocomposites were used to fabricate thin-film sensors. Of all the CNMs, the smallest limit of detection (LOD) was achieved for multi-walled CNT (MWCNT) with a LOD of 9.6 ppb for aminophenol and a very high linearity of 0.997, with an average sensitivity of 2.3 kΩ/pH at an acid pH. This high sensor performance can be attributed to the high homogeneity of the PANI coating on the MWCNT surface.

Ultrasensitive haptoglobin biomarker detection based on amplified chemiluminescence of magnetite nanoparticles.

BACKGROUND: Haptoglobin is an acute-phase protein used as predicting diagnostic biomarker both in humans (i.e., diabetes, ovarian cancer, some neurological and cardiovascular disorders) and in animals (e.g., bovine mastitis). The latter is a frequent disease of dairy industry with staggering economical losses upon decreased milk production and increased health care costs. Early stage diagnosis of the associated diseases or inflammation onset is almost impossible by conventional analytical manners. RESULTS: The present study demonstrates a simple, rapid, and cost-effective label-free chemiluminescence bioassay based on magnetite nanoparticles (MNPs) for sensitive detection of haptoglobin by employing the specific interaction of hemoglobin-modified MNPs. The resulting haptoglobin-hemoglobin complex inhibits the peroxidase-like activity of luminol/H2O2-hemoglobin-MNPs sensing scheme and reduces the chemiluminescence intensities correspondingly to the innate haptoglobin concentrations. Quantitative detection of bovine haptoglobin was obtained within the range of 1 pg mL-1 to 1 µg mL-1, while presenting 0.89 pg mL-1 limit of detection. Moreover, the influence of causative pathogenic bacteria (i.e., Streptococcus dysgalactiae and Escherichia coli) and somatic cell counts (depicting healthy, sub-clinical and clinical mastitis) on the emitted chemiluminescence radiation were established. The presented bioassay quantitative performances correspond with a standardized assay kit in differentiating dissimilar milk qualities. CONCLUSIONS: Overall, the main advantage of the presented sensing concept is the ability to detect haptoglobin, at clinically relevant concentrations within real milk samples for early bio-diagnostic detection of mastitis and hence adjusting the precise treatment, potentially initiating a positive influence on animals' individual health and hence on dairy farms economy.

Detection of trace heavy metal ions in water by nanostructured porous Si biosensors.

A generic biosensing platform, based on nanostructured porous Si (PSi), Fabry-Pérot thin films, for label-free monitoring of heavy metal ions in aqueous solutions by enzymatic activity inhibition, is described. First, we show a general detection assay by immobilizing horseradish peroxidase (HRP) within the oxidized PSi nanostructure and monitor its catalytic activity in real time by reflective interferometric Fourier transform spectroscopy. Optical studies reveal the high specificity and sensitivity of the HRP-immobilized PSi towards three metal ions (Ag(+) > Pb(2+) > Cu(2+)), with a detection limit range of 60-120 ppb. Next, we demonstrate the concept of specific detection of Cu(2+) ions (as a model heavy metal) by immobilizing Laccase, a multi-copper oxidase, within the oxidized PSi. The resulting biosensor allows for specific detection and quantification of copper ions in real water samples by monitoring the Laccase relative activity. The optical biosensing results are found to be in excellent agreement with those obtained by the gold standard analytical technique (ICP-AES) for all water samples. The main advantage of the presented biosensing concept is the ability to detect heavy metal ions at environmentally relevant concentrations using a simple and portable experimental setup, while the specific biosensor design can be tailored by varying the enzyme type.

Picking up the pieces: a generic porous Si biosensor for probing the proteolytic products of enzymes.

A multifunctional porous Si biosensor that can both monitor the enzymatic activity of minute samples and allow subsequent retrieval of the entrapped proteolytic products for mass spectrometry analysis is described. The biosensor is constructed by DNA-directed/reversible immobilization of enzymes onto a Fabry-Pérot thin film. We demonstrate high enzymatic activity levels of the immobilized enzymes (more than 80%), while maintaining their specificity. Mild dehybridization conditions allow enzyme recycling and facile surface regeneration for consecutive biosensing analysis. The catalytic activity of the immobilized enzymes is monitored in real time by reflective interferometric Fourier transform spectroscopy. The real-time analysis of minute quantities of enzymes (concentrations at least 1 order of magnitude lower, 0.1 mg mL(-1), in comparison to previous reports, 1 mg mL(-1)), in particular proteases, paves the way for substrate profiling and the identification of cleavage sites. The biosensor configuration is compatible with common proteomic methods and allows for a successful downstream mass spectrometry analysis of the reaction products.

SERS-based immunosensor for E. coli contaminants detection in milk using silver-coated nanoporous silicon substrates.

The dairy sector is frequently affected by contagious and environmental factors that spread between animals by numerous means and induce the inflammatory disease of bovine mastitis (BM). Herein, silver decorated porous silicon (Ag-pSi) SERS platform was designed for rapid and reliable Escherichia coli (predominant BM pathogen) detection in various milk origins. The inherent surface void and pore morphology were physically optimized to augment the SERS effect using 4-aminothiphenol (4ATP) while achieving an enhancement factor >4.6 × 107. An indirect immunoassay evaluated the residual unreacted antibodies using an optimized 4ATP/Ag-pSi SERS platform modified with secondary antibodies. Under optimized conditions, the porous substrate offered high sensitivity toward target bacteria detection of 3 CFU mL-1 and linear response of 101-105 CFU mL-1. Moreover, the selectivity and specificity of the designed sensing platform were cross-validated against other interfering bacteria without compromising its performance efficiencies. Finally, the applicability of the developed system for real-life conditions was elucidated in different milk samples (bovine, goat, sheep) with recovery values of 78-115% compared to the conventional culture technique. Considering the complex media analysis, the miniaturized SERS platform is highly reliable, rapid and accurate that could be applicable for routine on-site analysis of various emerging pathogens relevant to BM management.

Porous silicon Fabry-Pérot interferometer designed for sensitive detection of aflatoxin B1 in field crops.

Herein, a label-free sensing platform was designed for accurate, rapid and selective detection of aflatoxin B1 (AFB1), a potent mutagenic and carcinogenic substance in food and feedstuff. Minute AFB1 residues were assessed by competitive immunoassay facilitated on porous silicon Fabry-Pérot interferometer. The immunocomplex formation was biochemically amplified by enzymatic reaction products infiltrating the porous void and alternating the reflectivity spectra in correlation to the AFB1 content. The optical output presented high sensitivity toward target analyte detection in simulated conditions, as low as 0.03 ppb within the dynamic range of 0.01-10 ppb. The selectivity and specificity of the developed sensing platform were cross-validated versus commonly known interfering mycotoxins without compromising its performance values. Finally, the efficiency and the accuracy of the system were demonstrated in three matrices (maize, peanut and wheat) while demonstrating acceptable recovery values of 94-101 %, in compliance with the competitive ELISA standard assay and HPLC.

Enhancement of integrated nano-sensor performance comprised of electrospun PANI/carbonaceous material fibers for phenolic detection in aqueous solutions.

The detection of trace levels of organic residue in water samples is a key health issue. This manuscript describes the fabrication of integrated nano-sensors composed of electrospun microfibers consisting of a nanocomposite of carbonaceous materials (CNMs) containing polyaniline (PANI) and polycaprolactone (PCL) for phenolic detection in aqueous solutions. The morphology of the resulting microfiber composite was characterized by scanning electron microscopy. It revealed elongated fibers with a highly interconnected web-like pattern in the presence of reduced graphene oxide (rGO). Shorter microfibers were observed in the composite filled with multi-walled carbon nanotubes (MWCNTs), whereas large agglomerates were formed upon the incorporation of single-walled CNTs (SWCNTs) and graphene 300 (G300). Comparative analysis showed that the PANI/CNM sensors exhibited the best electrochemical properties, in particular in the presence of rGO and MWCNTs, where greater electrical conductivity was achieved, i.e., 4.33 × 10-3 and 7.22 × 10-4 S/cm, respectively, as compared to the PANI-PCL sensor (3.79 × 10-4 S/cm). All the PANI/CNM sensors exhibited high sensitivity. Notably, PANI/rGO was found to have a detection limit of 8.34 × 10-3 µM for aminophenol. All the sensors exhibited good selectivity in the presence of interference to detecting phenolic compounds in aqueous solutions, thus confirming their value for industrial applications.

Advancing nanostructured porous si-based optical transducers for label free bacteria detection.

Optical label-free porous Si-based biosensors for rapid bacteria detection are introduced. The biosensors are designed to directly capture the target bacteria cells onto their surface with no prior sample processing (such as cell lysis). Two types of nanostructured optical transducers based on oxidized porous Si (PSiO(2)) Fabry-Pérot thin films are synthesized and used to construct the biosensors. In the first system, we graft specific monoclonal antibodies (immunoglobulin G's) onto a neat electrochemically-machined PSiO(2) surface, based on well-established silanization chemistry. The second biosensor class consists of a PSiO(2)/hydrogel hybrid. The hydrogel, polyacrylamide, is synthesized in situ within the nanostructured PSiO(2) host and conjugated with specific monoclonal antibodies to provide the active component of the biosensor. Exposure of these modified-surfaces to the target bacteria results in "direct-cell-capture" onto the biosensor surface. These specific binding events induce predictable changes in the thin-film optical interference spectrum of the biosensor. Our studies demonstrate the applicability of these biosensors for the detection of low bacterial concentrations, in the range of 10(3)-10(5) cell/ml, within minutes. The sensing performance of the two different platforms, in terms of their stability in aqueous media and sensitivity, are compared and discussed. This preliminary study suggests that biosensors based on PSiO(2)/hydrogel hybrid outperform the neat PSiO(2) system.

N-acetyl-ß-d-glucosaminidase activity assay for monitoring insulin-dependent diabetes using Ag-porous Si SERS platform.

Determination of urinary or serum N-acetyl-ß-d-glucosaminidase (NAG) activity as a tissue damage indicator is widely used in diagnosis of various pathologies, including diabetic nephropathy. Early and rapid biomarker detection is an important element of medical diagnosis, facilitating prompt therapeutic decisions and prognosis evaluation. Herein, we present a modified sensing approach for a rapid and reliable NAG activity determination in complex media using surface-enhanced Raman spectroscopy (SERS). Porous silicon (PSi) Fabry-Pérot interferometers were redesigned as sensitive SERS platforms utilizing the vast inherent surface area for silver (Ag) nanoparticles embedment. Interaction of the porous nanostructures with specific NAG-enzymatic products produces an indicative spectral fingerprint proportional in magnitude to its concentration. The sensitivity of Ag-PSi SERS substrates was evaluated in complex matrices presenting sufficient limits of detection compared with other advanced assays and techniques (0.07, 0.47 and 0.50 mU mL-1 for urine, milk and plasma, respectively). The augmented optical performance revealed recovery values of 96-109%, indicating successful and selective NAG recognition in biological fluids. Finally, the potential applicability of the suggested prototype for real-life scenarios was evaluated in vivo, in a model of insulin-dependent diabetes induced in sheep. Overall, the robust data confirm the application of SERS analysis for early diagnosis of pathology and for evaluation of clinical responses to pharmacological treatments.

Engineering nanostructured porous SiO2 surfaces for bacteria detection via "direct cell capture".

An optical label-free biosensing platform for bacteria detection ( Escherichia coli K12 as a model system) based on nanostructured oxidized porous silicon (PSiO(2)) is introduced. The biosensor is designed to directly capture the target bacteria cells on its surface with no prior sample processing (such as cell lysis). The optical reflectivity spectrum of the PSiO(2) nanostructure displays Fabry-Pérot fringes characteristic of thin-film interference, enabling direct, real-time observation of bacteria attachment within minutes. The PSiO(2) optical nanostructure is synthesized and used as the optical transducer element. The porous surface is conjugated with specific monoclonal antibodies (immunoglobulin G's) to provide the active component of the biosensor. The immobilization of the antibodies onto the biosensor system is confirmed by attenuated total reflectance Fourier transform infrared spectroscopy, fluorescent labeling experiments, and refractive interferometric Fourier transform spectroscopy. We show that the immobilized antibodies maintain their immunoactivity and specificity when attached to the sensor surface. Exposure of these nanostructures to the target bacteria results in "direct cell capture" onto the biosensor surface. These specific binding events induce predictable changes in the thin-film optical interference spectrum of the biosensor. Our preliminary studies demonstrate the applicability of these biosensors for the detection of low bacterial concentrations. The current detection limit of E. coli K12 bacteria is 10(4) cells/mL within several minutes.

Bovine mastitis inflammatory assessment using silica coated ZnO-NPs induced fluorescence of NAGase biomarker assay.

Bovine mastitis (BM) is the most common inflammatory disease in the dairy sector worldwide, originated from bacterial invasion onto the mammary gland. Early BM detection is crucial for identifying new pathogenic infections within the dairy herd, which can be alleviated by antimicrobial therapy. N-acetyl-ß-D-glucosaminidase (NAGase) is a prominent BM inflammatory biomarker secreted onto the blood circulation upon pathogenesis and then released into milk, capable of separating healthy quarters from subclinical and clinical BM cases. Herein, we report on a sensitive differentiation assay of BM severity based on enhanced fluorescence emission of a conventional NAGase activity assay. The addition of silica-coated zinc oxide nanoparticles induces non-radiative energy transfer to the lysosomal reaction products, thus leading to enhanced fluorescence (above 3-fold). Various milk qualities within the entire inflammatory spectrum were evaluated by the modified fluorescence assay with respect to non-infected milk. The amplified emission values differentiate between two predominant BM causative pathogens (Streptococcus dysgalactiae and Escherichia coli) at various somatic cell counts. In general, the presented concept offers an efficient, simple, cost-effective fluorescence signal augmentation for mastitis identification, thus offering means to diagnose the severity of the associated disease.

Amplified Fluorescence by ZnO Nanoparticles vs. Quantum Dots for Bovine Mastitis Acute Phase Response Evaluation in Milk.

Bovine mastitis (BM) is a prominent inflammatory disease affecting the dairy industry worldwide, originated by pathogenic agent invasion onto the mammary gland. The early detection of new BM cases is of high importance for infection control within the herd. During inflammation, various biomarkers are released into the blood circulation, which are consequently found in milk. Herein, the lysosomal activity of N-acetyl-ß-D-glucosaminidase (NAGase), a predominant BM indicator, was utilized for highly sensitive clinical state differentiation. The latter is achieved by the precise addition of tetraethyl orthosilicate-coated zinc oxide nanostructures (quantum dots or nanoparticles, individually) onto a conventional assay. Enhanced fluorescence due to the nanomaterial accumulative near-field effect is achieved within real milk samples, contaminated with Streptococcus dysgalactiae, favoring quantum dots over nanoparticles (> 7-fold and 3-fold, respectively), thus revealing significant differentiation between various somatic cell counts. The main advantage of the presented sensing concept, besides its clinically relevant concentrations, is the early bio-diagnostic detection of mastitis (subclinical BM) by using a simple and cost-effective experimental setup. Moreover, the assay can be adapted for BM recovery prognosis evaluation, and thus impact on udder health status, producing an alternative means for conventional diagnosis practices.

Inflammatory biomarker detection in milk using label-free porous SiO2 interferometer.

N-acetyl-ß-d-glucosaminidase (NAGase) is an established indicative biomarker released upon damage or necrosis of tubular epithelial cells in both humans and animals, indicating severe nephrological disorders and bovine mastitis (BM), respectively. The latter is the most common and costly disease in dairy cattle associated with production losses, elevated somatic cell counts and deteriorated health status. Herein, we report on a reflective based assay for early diagnosis of BM through the analysis of NAGase inherent content found in whole milk samples using a miniaturized optical transducer. Gelatin functionalized porous Si Fabry-Pérot interferometers are employed for monitoring the lysosomal activity in various stages of the inflammation (healthy, subclinical and clinical). The enzymatic reaction products precipitate and accumulate within the porous nanostructure, thus alter the average refractive index monitored using reflectometric interference spectroscopy. The optical assay is calibrated within the clinically relevant concentrations of BM while presenting a dynamic range of 1.04-16.7 µM min-1 and the detection limit of 0.49 µM min-1. The specific optical performance of the biosensor correlates with a gold standard laboratory-based approach, in which escalated somatic cell counts reveal augmented NAGase levels and thus severe pathogenesis. Overall, our study provides new opportunities to develop a convenient bio-diagnostic sensing system for BM detection and classification by addressing the limitations of conventional practices.

Porous Silicon Fabry-Pérot Interferometer for N-Acetyl-ß-d-Glucosaminidase Biomarker Monitoring.

Bovine mastitis (BM) is a prominent inflammatory disease affecting the dairy industry worldwide, originated by pathogenic agent invasion onto the mammary gland. Early detection of new BM cases is of high importance for infection control within the herd. Conventional analytical techniques lack the ability to detect BM-predicting biomarkers, used as analytical indicators for health status evaluation, in real time or outside the laboratory boundaries. Herein, we describe a biosensing platform for label-free detection and identification of BM onset through targeting N-acetyl-ß-d-glucosaminidase (NAGase) for potential evidence-based therapy. The lysosomal activity in dissimilar milk qualities was monitored by a gelatin-functionalized porous Si Fabry-Pérot interferometer, while estimating the biochemical reaction precipitating products within the nanostructure. The optical response was proportional to the inherent NAGase concentration found in real milk samples, influenced by two dominant BM causative pathogens (i.e., Escherichia coli and Streptococcus dysgalactiae) at various somatic cell counts. Quantitative analysis of NAGase levels within the entire inflammatory spectrum (healthy, subclinical, and clinical BM) was obtained within the range of 1.0-4.2 µM/min (enzymatic activity per volume unit), while presenting a detection limit of 0.51 µM/min. The optical performances correspond with standardized biochemical activity assay in dissimilar milk qualities. Overall, the presented sensing concept exhibits the potential of BM-predicting biomarker detection using a simple and portable experimental setup for convenient early biodiagnostics and health status evaluation.

Gold Nanoparticle Size-Dependent Enhanced Chemiluminescence for Ultra-Sensitive Haptoglobin Biomarker Detection.

Bovine mastitis (BM) is a frequent disease in the dairy industry that causes staggering economical losses due to decreased milk production and increased health care costs. Traditionally, BM detection depends on the efficacy and reliability of analytical techniques that measure somatic cell counts (SCC), detect pathogens, and reveal inflammatory status. Herein, we demonstrate the detection of bovine haptoglobin, a well-documented acute phase protein for evaluating BM clinical status, by utilizing hemoglobin-binding capacity within luminol chemiluminescence (CL) system. The resulting haptoglobin-hemoglobin complex reduces the CL signal proportionally to inherent haptoglobin concentrations. Different sizes of cross-linked gold nanoparticles (GNPs) were examined for enhanced CL (eCL) signal amplification, presenting over 30-fold emitted radiation enhancement for optimized size within real milk samples with respect to nanoparticle-free assay. The eCL values were proportionally related to nanoparticle size and content, influenced by SCC and pathogen type (e.g., Escherichia coli and coagulase-negative staphylococci). The optimized bioassay showed a broad linear response (1 pg mL-1-10 µg mL-1) and minute detection limit of 0.19 pg mL-1, while presenting quantitative performance in agreement with commercial ELISA kit. Finally, the resulting optimized eCL concept offers an efficient label-free detection of haptoglobin biomarker, offering means to diagnose the severity of the associated diseases.

Enhanced Fluorescence of N-Acetyl-ß-D-Glucosaminidase Activity by ZnO Quantum Dots for Early Stage Mastitis Evaluation.

Recurrent mastitis events are the major cause of annual revenue losses in the dairy sector resulting in decreased milk yield, escalading treatment costs and increased health risk of the entire herd. Upon udder inflammation, several biomarkers are proportionally secreted to its severity onto the blood circulation and consequently into milk (upon breached blood-milk barrier). N-acetyl-ß-D-glucosaminidase activity is widely used mastitis indicator in milk, offering simple means of differentiation between healthy quarters from those with subclinical or clinical severity. Herein, we demonstrate fluorescence signal amplification concept for sensitive clinical status discrimination. Tetraethyl orthosilicate coated zinc oxide quantum dots were employed within the conventional N-acetyl-ß-D-glucosaminidase activity assay. Under the experimental conditions, a profound non-radiative energy transfer occurred between quantum nanomaterials onto enzymatic fluorescent products resulting in intensified emission of the latter, over 11-folds, in comparison to nanoparticle-free assay. Overall, the fluorescence intensities were proportionally related to zinc oxide quantum dots surface coverage and concentration, SCC values and influenced by the causing bacteria (i.e., Streptococcus dysgalactiae and Coagulase-negative Staphylococci). Finally, the presented proof-of-concept offers an efficient, simple, cost-effective fluorescence signal amplification for early stage mastitis identification, offering means to diagnose the severity of the associated diseases and hence deducing on animals' clinical status.

Milk haptoglobin detection based on enhanced chemiluminescence of gold nanoparticles.

The suggested research specifically addresses the major source of economic loss of the dairy industry, the bovine mastitis (BM), an inflammatory disease of mammary gland caused by bacterial intramammary infection. During udder inflammation, the concentrations of acute phase proteins (APP) in both plasma and milk are escalated, which can be distinctively utilized as predicting diagnostic biomarkers of cattle's BM clinical status. Herein, we demonstrate a liquid-phase luminol chemiluminescence (CL) system for sensitive detection of haptoglobin (Hp), a predictive APP of BM, by utilizing the binding capacity of hemoglobin (Hb). The CL intensity is linearly proportional to Hb-Hp complex formation, resulting in peroxidase-like activity inhibition of luminol-H2O2-Hb CL system. Enhanced CL, at least 10-fold effect within real samples, is attained by the addition of catalytically active cross-linked gold nanoparticles (GNPs) onto the luminol-H2O2 solution. Moreover, the influence of different somatic cell counts (representing subclinical and clinical BM status) and pathogen types (i.e., CNS and Streptococcus dysgalactiae) on the secreted milk Hp levels obtained from Holstein cows are established. The analyzed Hp concentrations are in agreement with a commercial enzyme-linked immunosorbent assay kit. The proposed CL sensing concept offers cost-effective, simple, label-free and reliable systematic analysis of Hp biomarker for BM, potentially initiating a positive effect on animals' health and overall economy of the dairy farms.

A Novel Intra-aortic Device Designed for Coronary Blood Flow Amplification in Unrevascularizable Patients.

Patients with unrevascularizable coronary artery disease represent a substantial number of all patients with coronary disease. However, their therapeutic options are limited; they endure recurrent hospitalizations, a poor quality of life and prognosis. We aim to investigate a novel alternative approach to the treatment of this common medical condition by using a specialized intra-aortic device with coiling properties capable of enhancing diastolic coronary flow. Both a mathematical analysis and in vitro study presented in the current study have yielded enhanced coronary diastolic blood flow and energetic advantages. We suggest that this original approach might be implicated in severely symptomatic unrevascularizable patients.