Multimodal diagnosis of cerebrospinal fluid rhinorrhea: State of the art review and emerging concepts.

Objective: Currently, diagnosis of cerebrospinal fluid (CSF) rhinorrhea relies on a multimodal approach, increasing costs and ultimately delaying diagnosis. In the United States and internationally, the crux of such a diagnosis relies on confirmation testing (via biomarkers) and localization (e.g., imaging). Biomarker testing may require analysis at an outside facility, resulting in delays diagnosis and treatment. In addition, specialized imaging may be nonspecific and often requires an active leak for diagnosis. There remains a clear need for innovative new technology. Methods: A comprehensive review was conducted on both foundational and innovative scholarly articles regarding current and emerging diagnosis modalities for CSF. Results: Current modalities in CSF rhinorrhea diagnosis and localization include laboratory tests (namely, B2T immunofixation), imaging (CT and/or MRI) with or without intrathecal administration, and surgical exploration. Each of these modalities carry flaws, risks, and benefits, ultimately contributing to delays in diagnosis and morbidity. Promising emerging technologies include lateral flow immunoassays (LFI) and biologically functionalized field-effect transistors (BioFET). Nevertheless, these carry some drawbacks of their own, and require further validation. Conclusion: CSF rhinorrhea remains a challenging diagnosis, requiring a multimodal approach to differentiate from nonpathologic causes of rhinorrhea. Current methods in diagnosis are imperfect, as the ideal test would be a readily accessible, inexpensive, rapid, highly accurate point-of-care test without the need for excess fluid or specialized processing. Critical work is being done to develop promising, new, improved tests, though a clear successor has not yet emerged. Level of Evidence: N/A.

Chitosan Nanoparticles Loaded with Quercetin and Valproic Acid: A Novel Approach for Enhancing Antioxidant Activity against Oxidative Stress in the SH-SY5Y Human Neuroblastoma Cell Line.

BACKGROUND: Multiple drug-delivery systems obtained by loading nanoparticles (NPs) with different drugs that have different physicochemical properties present a promising strategy to achieve synergistic effects between drugs or overcome undesired effects. This study aims to develop a new NP by loading quercetin (Que) and valproic acid (VPA) into chitosan. In this context, our study investigated the antioxidant activities of chitosan NPs loaded with single and dual drugs containing Que against oxidative stress. METHOD: The synthesis of chitosan NPs loaded with a single (Que or VPA) and dual drug (Que and VPA), the characterization of the NPs, the conducting of in vitro antioxidant activity studies, and the analysis of the cytotoxicity and antioxidant activity of the NPs in human neuroblastoma SH-SY5Y cell lines were performed. RESULT: The NP applications that protected cell viability to the greatest extent against H2O2-induced cell damage were, in order, 96 µg/mL of Que-loaded chitosan NP (77.30%, 48 h), 2 µg/mL of VPA-loaded chitosan NP (70.06%, 24 h), 96 µg/mL of blank chitosan NP (68.31%, 48 h), and 2 µg/mL of Que- and VPA-loaded chitosan NP (66.03%, 24 h). CONCLUSION: Our study establishes a successful paradigm for developing drug-loaded NPs with a uniform and homogeneous distribution of drugs into NPs. Chitosan NPs loaded with both single and dual drugs possessing antioxidant activity were successfully developed. The capability of chitosan NPs developed at the nanometer scale to sustain cell viability in SH-SY5Y cell lines implies the potential of intranasal administration of chitosan NPs for future studies, offering protective effects in central nervous system diseases.

Electrochemistry Test Strip as Platform for In Situ Detection of Blood Levels of Antipsychotic Clozapine in Finger-Pricked Sample Volume.

With the increasing number of patients suffering from Parkinson's disease, the importance of measuring drug levels in patient body fluids has increased exponentially, particularly for the drug clozapine. There is a growing demand for real-time analysis of biofluids on a single low-cost platform in ultra-low fluid volumes with robustness. This study aims to measure the level of clozapine (Clz) with a portable potentiostat using a practical approach. For this purpose, we developed an inexpensive, portable platform via electrochemistry on a commercial glucose test strip (CTS). CTSs were first modified by removing the enzyme mixture from the surface of the sensing zone, which was followed by modification with Multi walled carbon nanotube (MWCNT) and Nafion. The electrochemical characteristics of CTS electrodes were investigated using cyclic voltammetry (CV) and differential voltammetry (DPV) techniques. The designed sensor displayed decent linear range, detection limit, reproducibility, and reusability results. A linear dynamic range of 0.1-5 µM clozapine was observed under optimized conditions with a good sensitivity (1.295 µA/µM) and detection limit (83 nM). Furthermore, the designed sensing electrode was used to measure the amount of Clz in real samples.

Polyamidoamine Dendron-Bearing Lipids as Drug-Delivery Excipients.

An amine-terminated polyamidoamine (PAMAM) dendron and two long alkyl groups were designed as a novel drug carrier that possesses an interior for the encapsulation of drugs and a biocompatible surface. We synthesized three dendron-bearing lipids, DL-G1, DL-G2, and DL-G3, which included first, second, and third generation polyamidoamine dendrons, respectively. The synthesized dendrimer encapsulating anticancer drug, 5-fluorouracil (5-FU), was prepared by extraction with chloroform from mixtures of the dendrimers and varying amounts of the drug. In vitro cytotoxicity of PAMAM conjugated di-n-dodecylamine micelles (G1, G2, G3) were analyzed on human gastric adenocarcinoma cells (AGS) by water-soluble tetrazolium-1 (WST-1) cell proliferation assay. Upon exposure to 5-FU loaded micelles, the viability of the cells decreased gradually in all generations. Cytotoxicity increased with increasing generation and reached its highest rate of 69.8 ± 3.2% upon 15 µM 5FU-loaded 25 µM PAMAM DL-3 micelle treatment. These results demonstrate that 5FU-loaded PAMAM conjugated di-n-dodecylamine treatment inhibits the proliferation of AGS cells in a generation-dependent manner.

Polyelectrolyte Multilayers Composed of Polyethyleneimine-Grafted Chitosan and Polyacrylic Acid for Controlled-Drug-Delivery Applications.

In this work, polyethyleneimine (PEI)-grafted chitosan (Chi-g-PEI) was prepared for the fabrication of layer-by-layer (LBL) films for use in sustained-drug-delivery applications. Chi-g-PEI and polyacrylic acid (PAA) multilayer films were formed using the LBL technique. Methylene blue (MB) was used as a model drug for the investigation of loading and release capabilities of the LBL films. Characterizations of the synthesized copolymer were performed using Fourier-transform infrared spectroscopy (FTIR), Nuclear magnetic resonance spectroscopy (NMR), Thermogravimetric analysis (TGA), and X-ray Powder Diffraction (XRD) techniques, and the thickness of the LBL films was measured using Atomic force microscopy (AFM). The drug-loading and -release behaviors of the LBL films were assessed using a UV-visible spectrophotometer. The results showed that the loading capacity and release rate of MB were affected by ionic strength and pH. In addition, it was demonstrated that PEI-grafted chitosan is a good candidate for the assembling of LBL films for drug-delivery applications.

Lab-In-A-Syringe: A Novel Electrochemical Biosensor for On-Site and Real-Time Monitoring of Dopamine in Freely Behaving Mice.

There is a growing demand for real-time analysis and sampling of biofluids on a single low-cost platform in ultralow fluid volumes with robustness. In this study, a microfluidic sensor was developed, manufactured through an additive manufacturing technique, and used for dopamine (DA) measurements. We implemented a biosensing system using pencil graphites (PGEs) integrated into a three-dimensional (3D) printed microfluidic syringe-type device (µSyringe). The amperometry technique was used to monitor the current changes associated with the electrooxidation of DA. The sensing signal was stable and linear in a concentration range of DA between the limit of quantification (0.1 nM) and the upper limit of linearity (500 nM). The µSyringe sensing device is simple, robust, and stable, making it suitable for real-time measurement of DA in cerebrospinal fluid (CSF) from freely moving mice.

Surface Plasmon Resonance Identifies High-Affinity Binding of l-DOPA to Siderocalin/Lipocalin-2 through Iron-Siderophore Action: Implications for Parkinson's Disease Treatment.

l-3,4-Dihydroxyphenylalanine (l-DOPA), the dopamine precursor, remains the frontline treatment for Parkinson's disease (PD). With the treatment progress, l-DOPA efficacy decreases, necessitating higher and more frequent doses, with higher risks of dyskinesia. l-DOPA chelates iron through its catechol group, forming the l-DOPA:Fe complex; however, the fate of this complex is unknown. Catechol siderophore-like compounds are known to bind siderocalin (Scn)/lipocalin-2 to form stable siderophore:Fe:Scn complexes. Scn is upregulated in PD patients' substantia nigra and may play a role in PD pathophysiology. Therefore, in this study, we used the surface plasmon resonance (SPR) technique to examine the binding properties of l-DOPA to Scn. We found that l-DOPA formed a stable complex with Scn in the presence of Fe3+. Our analysis of the binding properties of l-DOPA precursors and metabolites indicates that the catechol group is necessary but not sufficient to form a stable complex with Scn. Finally, the affinity constant (Kd) of DOPA:Fe3+ binding with Scn (0.8 µM) was lower than l-DOPA plasma peak concentrations in l-DOPA preparations in the past six decades. Our results speculate a significant role for the l-DOPA-Scn complex in the decreased bioavailability of l-DOPA with the progress of PD.

Lab-in-a-pencil graphite: A 3D-printed microfluidic sensing platform for real-time measurement of antipsychotic clozapine level.

A novel lab-in-a-pencil graphite microfluidic sensing electrode (µFSE) was fabricated for real-time flow injection measurement of the antipsychotic drug clozapine (Clz). A simple, low-cost, and reusable µFSE was obtained by using 3D printing of a microfluidic chamber integrated with a flat pencil graphite without the need to utilize complex technologies. The µFSE has tubular geometry with 800 µm diameter, where the solution continuously flows in the holes of flat pencil graphite electrodes. Under optimized conditions, this device offers fast and effective Clz detection with good analytical features. A linear calibration curve in the range of 0.5 to 10 µM Clz was obtained with good sensitivity (0.01275 µA µM-1) and detection limit (24 nM). Finally, we demonstrate the applicability of our lab-fabricated microfluidic electrochemical device by monitoring Clz in serum samples at low concentrations.

Vorinostat-loaded titanium oxide nanoparticles (anatase) induce G2/M cell cycle arrest in breast cancer cells via PALB2 upregulation.

Breast cancer is a group of diseases in which cells divide out of controlled, typically resulting in a mass. Erlotinib is targeted cancer drug which functions as an inhibitor of the epidermal growth factor receptor (EGFR) tyrosine kinase. It is used mainly to treat of non-small cell lung cancer patients and has an action against pancreatic cancer. Vorinostat (aka suberanilohydroxamic acid) is an inhibitor of histone deacetylases (HDAC), which has an epigenetic modulation activity. It is used to treat cutaneous T cell lymphoma. In the present study, the erlotinib (ERL) and vorinostat (SAHA) loaded TiO2 nanoparticles (NPs) were used for the treatment of the breast cancer cells (MDA-MB-231 and MCF-7) and human cancerous amniotic cells (WISH). Cell count and viability were negatively affected in all treatments compared to normal cells and bare TiO2 NPs. Apoptosis results indicated a significant increase in the total apoptosis in all treatments compared with control cells. ERL- and SAHA-loaded TiO2 NPs treatments arrested breast cancer cells at G2/M phase, which indicate the cytotoxic effect of these treatment. Partner and localizer of BRCA2 (PALB2) gene expression was assessed using qPCR. The results indicate that PLAB2 was upregulated in ERL- and SAHA-loaded TiO2 NPs compared with control cells and can be used as nanocarrier for chemotherapy drugs. However, this conclusion necessitates further confirmative investigation.

Microfluidic Electrochemical Sensor for Cerebrospinal Fluid and Blood Dopamine Detection in a Mouse Model of Parkinson's Disease.

Parkinson's disease (PD) is a progressive neurodegenerative disorder involving dopaminergic neurons from the substantia nigra. The loss of dopaminergic neurons results in decreased dopamine (DA) release in the striatum and thus impaired motor functions. DA is one of the key neurotransmitters monitored for the diagnosis and during the progression and treatment of PD. Therefore, sensitive and selective DA detection methods are of high clinical relevance. In this study, a new microfluidic device utilized for electrochemical DA detection is reported. The microfluidic sensing device operates in the range of 0.1-1000 nM DA requiring only ∼2.4 µL sample volume, which corresponds to detectable 240 amol of DA. Using this sensor, we were able to monitor the changes in DA levels in cerebrospinal fluid and plasma of a mouse model of PD and following the treatment of drug l-3,4-dihydroxyphenylalanine.

Electrochemical Micropyramid Array-Based Sensor for In Situ Monitoring of Dopamine Released from Neuroblastoma Cells.

Abnormal dopamine neurotransmission is associated with several neurological and psychiatric disorders such as Parkinson's disease, schizophrenia, attention deficiency and hyperactivity disorder, and addiction. Developing highly sensitive, selective, and fast dopamine monitoring methods is of high importance especially for the early diagnosis of these diseases. Herein, we report a new ultrasensitive electrochemical sensing platform for in situ monitoring of cell-secreted dopamine using Au-coated arrays of micropyramid structures integrated directly into a Petri dish. This approach enables the monitoring of dopamine released from cells in real-time without the need for relocating cultured cells. According to the electrochemical analyses, our dopamine sensing platform exhibits excellent analytical characteristics with a detection limit of 0.50 ± 0.08 nM, a wide linear range of 0.01-500 µM, and a sensitivity of 0.18 ± 0.01 µA/µM. The sensor also has remarkable selectivity toward DA in the presence of different potentially interfering small molecules. The developed electrochemical sensor has great potential for in vitro analysis of neuronal cells as well as early diagnosis of different neurological diseases related to abnormal levels of dopamine.

Enhanced electrochemical sensing performance by insitu electrocopolymerization of pyrrole and thiophene-grafted chitosan.

Nowadays, there is increasing number of electrochemical biosensors which utilize chitosan (Ch); as an enzyme immobilization matrix, and conductive nanomaterials; as electron carriers improving sensitivity of the biosensor. However, the challenge these sensors face is the lack of uniform dispersion of nanomaterials throughout the Ch film, which can negatively affect analytical performance of the biosensor. In this study, we report the development of an enzyme immobilization matrix that displays enhanced electrochemical performance thanks to a novel conductive thin film prepared via in situ electrocopolymerization of pyrrole (Py) and thiophene-grafted chitosan (Th-Ch). This is a simple thin film preparation method that can help overcome aforementioned challenges by providing a uniformly distributed conductive layer on the electrode. We are also for the first time reporting the synthesis and characterization of Th-Ch, where grafted Th plays an essential role as a linking group between Ch and Py. The resulting conductive Ch-based thin film was modified with glucose oxidase (GOx) which served as a model enzyme. In situ electrocopolymerization of Py with Th-Ch resulted in a highly conductive thin film enabling approximately 40% higher sensitivity when compared to a Py-Ch composite. This new type of composite thin film is promising in biosensor technology due to its biocompatibility, the chemically and physically modifiable structure, as well as its electrical conductivity.

Recent progress in nanomaterial-based electrochemical and optical sensors for hypoxanthine and xanthine. A review.

This review (with 160 ref.) summarizes the progress that has been made in the methods for chemical or biochemical sensing of hypoxanthine and xanthine, which are produced as part of purine metabolism and are precursors of uric acid. An introduction discusses the importance of hypoxanthine and xanthine as analytes due to their significance in the clinical and food science, together with the conventional methods of analysis. A large section covers methods for the electrochemical hypoxanthine and xanthine sensing. It is divided into subsections according to the nanomaterials used including carbon nanomaterials, meal oxide nanoparticles, metal organic frameworks, conductive polymers, and bio-nanocomposites. A further large section covers optical methods for hypoxanthine and xanthine sensing, with subsections on nanomaterials including carbon nanomaterials, nanosheets, nanoclusters, nanoparticles, and their bio-nanocomposites. A concluding section summarizes the current status, addresses current challenges, and discusses future perspectives. Graphical abstractSchematic representation of the hypoxanthine and xanthine electrochemical and optical sensors incorporating various nanomaterials like graphene, carbon nanotubes (CNT), quantum dots (QD), nanoparticles and polymers, which are implemented in clinical and food analysis.

Electrochemical DNA biosensors for label-free breast cancer gene marker detection.

We present an electrochemical DNA detection strategy based on self-assembled ferrocene-cored poly(amidoamine) dendrimers for the detection of a gene relevant to breast cancer. The chemisorption of three ferrocene-cored poly(amidoamine) generations and hybridization of single-stranded DNA on a Au electrode were studied by cyclic voltammetry and differential pulse voltammetry. The biosensor demonstrated high sensitivity of 0.13 µA/(ng/ml) in the detection of the target DNA with a linear range of 1.3-20 nM and a detection limit of 0.38 nM. The DNA biosensor also has high selectivity for the target DNA, showing a clear signal difference from a noncomplementary sequence and a single-base-mismatch sequence, which was used as a model of BRAC1 gene mutation. The results shown are highly motivating for exploring DNA biosensing technology in the diagnosis of breast cancer caused by mutation of the BRAC1 gene. Graphical abstract.

Construction of conducting polymer/cytochrome C/thylakoid membrane based photo-bioelectrochemical fuel cells generating high photocurrent via photosynthesis.

In this study, a photo-bioelectrochemical fuel cell was constructed for photocurrent generation by illuminating the electrodes within an aqueous solution. In this purpose, gold electrode was coated with poly 4-(4H-Dithieno [3,2-b:2',3'-d]pyrol-4-yl) aniline, P(DTP-Ph-NH2) conductive polymer film by using electrochemical polymerization. Then, P(DTP-Ph-NH2) conductive polymer film coated surface was electrochemically modified with cytochrome C which covalently linked onto the surface via bis-aniline functionality of the polymer film and formed crosslinked-structure. The thylakoid membrane was attached on the surface of this electrode by using bissulfosaxinimidyl suberate (BS3) and used as photo-anode in photo-bioelectrochemical fuel cell. The photo-cathode of the photo-bioelectrochemical fuel cell fabrication was followed by the modification of conductive polymer poly[5-(4H-dithieno [3,2-b:2',3'-d]pyrol-4-yl) naphtalene-1-amine] film coating, glutaraldehyde activation, and bilirubin oxidase enzyme immobilization. During the photosynthesis occurring in thylakoid membrane under the light, water was oxidized and separated; while oxygen was released in anode side, the cathode side was reduced the oxygen gas into the water via a bio-electro-catalytic method. The cytochrome C was used for binding of thylakoid membrane to the electrode surface and play an important role for transferring of electrons released as a result of photosynthesis.

Novel electrochemical xanthine biosensor based on chitosan-polypyrrole-gold nanoparticles hybrid bio-nanocomposite platform.

The aim of this study was the electrochemical detection of the adenosine-3-phosphate degradation product, xanthine, using a new xanthine biosensor based on a hybrid bio-nanocomposite platform which has been successfully employed in the evaluation of meat freshness. In the design of the amperometric xanthine biosensor, chitosan-polypyrrole-gold nanoparticles fabricated by an in situ chemical synthesis method on a glassy carbon electrode surface was used to enhance electron transfer and to provide good enzyme affinity. Electrochemical studies were carried out by the modified electrode with immobilized xanthine oxidase on it, after which the biosensor was tested to ascertain the optimization parameters. The Biosensor exhibited a very good linear range of 1-200 µM, low detection limit of 0.25 µM, average response time of 8 seconds, and was not prone to significant interference from uric acid, ascorbic acid, glucose, and sodium benzoate. The resulting bio-nanocomposite xanthine biosensor was tested with fish, beef, and chicken real-sample measurements.

Construction of ferrocene modified conducting polymer based amperometric urea biosensor.

Herein, an electrochemical urea sensing bio-electrode is reported that has been constructed by firstly electropolymerizing 4-(2,5-Di(thiophen-2-yl)-1H-pyrrol-1-yl)aniline monomer (SNS-Aniline) on Pencil Graphite Electrode (PGE), then modifying the polymer coated electrode surface with di-amino-Ferrocene (DAFc) as the mediator, and lastly Urease enzyme through glutaraldehyde crosslinking. The effect of pH, temperature, polymer thickness, and applied potential on the electrode current response was investigated besides performing storage and operational stability experiments with the interference studies. The resulting urea biosensor's amperometric response was linear in the range of 0.1-8.5mM with the sensitivity of 0.54µA/mM, detection limit of 12µM, and short response time of 2s. The designed bio-electrode was tested with real human blood and urine samples where it showed excellent analytical performance with insignificant interference.

An electrochemical immunosensor for sensitive detection of Escherichia coli O157:H7 by using chitosan, MWCNT, polypyrrole with gold nanoparticles hybrid sensing platform.

An electrochemical immunosensor for the common food pathogen Escherichia coli O157:H7 was developed. This novel immunosensor based on the PPy/AuNP/MWCNT/Chi hybrid bionanocomposite modified pencil graphite electrode (PGE). This hybrid bionanocomposite platform was modified with anti-E. coli O157:H7 monoclonal antibody. The prepared bionanocomposite platform and immunosensor was characterized by using cyclic voltammetry (CV). Under the optimum conditions, the results have shown the order of the preferential selectivity of the method is gram negative pathogenic species E. coli O157:H7. Concentrations of E. coli O157:H7 from 3×101 to 3×107cfu/mL could be detected. The detection limit was ∼30cfu/mL in PBS buffer. Briefly, we developed a high sensitive electrochemical immunosensor for specific detection of E. coli O157:H7 contamination with the use of sandwich assay evaluated in this study offered a reliable means of quantification of the bacteria. For the applications in food quality and safety control, our immunosensor showed reproducibility and stability.

Dendrimer functional hydroxyapatite nanoparticles generated by functionalization with siloxane-cored PAMAM dendrons.

The surface modification of nano-hydroxyapatite with polyamidoamine (PAMAM) dendrimer has potential for biomedical applications due to their unique and well-defined secondary structures. This paper presents a facile method to synthesize siloxane-cored PAMAM dendrons modified Hydroxyapatite nanoparticles. Firstly, the siloxane-cored PAMAM dendrons with different generations (G1 to G5, respectively) were synthesized using repetitive reactions between Michael addition and amidation starting from 3-aminopropyltriethoxysilane. Then, hydroxyapatite nanoparticles were synthesized by hydrothermal crystallization method and functionalized with siloxane-cored PAMAM dendrons. These synthesized materials were characterized by using Fourier-transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), Scanning electron microscopy (SEM) and Thermogravimetric analysis (TGA). The results showed that the PAMAM dendrimer functionalization was carried out successfully and these materials may be applicable in biocomposite material and/or bone tissue engineering.

Preparation and characterization of amine functional nano-hydroxyapatite/chitosan bionanocomposite for bone tissue engineering applications.

In this study, three different types of scaffolds including a uniquely modified composite scaffold - namely chitosan (CTS), nano-hydroxyapatite/chitosan composite (CTS+nHAP), and amine group (NH2) modified nano-hydroxyapatite/chitosan composite (CTS+nHAP-NH2) scaffolds - were synthesized for bone tissue engineering (BTE) purposes. As results of the study, it was found that all scaffold types were biodegradable with CTS and CTS+nHAP scaffolds losing up to 15% of their initial weight, while the CTS+nHAP-NH2 scaffold showing 10% of weight loss after six weeks of lysozyme treatment. In addition, all three types of scaffolds were shown to be biocompatible, and amongst them CTS+nHAP-NH2 scaffolds supported the most cell proliferation in WST-1 assay and expressed the least and acceptable level of cytotoxicity in lactate dehydrogenase (LDH) test for human bone mesenchymal stem cells (hBM-MSCs). Finally, during osteoinductivity assessment, CTS+nHAP-NH2 nearly tripled initial alkaline phosphatase (ALP) activity when whereas both CTS and CTS+nHAP scaffolds only doubled. These results indicate that all synthesized scaffold types under investigation have certain potential to be used in bone tissue engineering approaches with CTS+nHAP-NH2 scaffold being the most promising and applicable one. In the future, we plan to intensify our studies on osteogenic differentiation on our scaffolds on a detailed molecular level and to include in vivo studies for pre-clinical purposes.