A promising 'single' and 'dual' drug-nanocomposite enriched contact lens for the management of glaucoma in response to the tear fluid enzyme.

Glaucoma is a neurodegenerative condition that results in the damage of retinal ganglion cells due to elevated intraocular pressure (IOP). To curtail the limitations associated with conventional treatments such as eye drops and ocular suspensions, we have developed 'single' and 'dual' drug delivery contact lenses (CLs), that is, latanoprost (LP) and latanoprost-timolol (LP-TM) deliverable CLs, in response to lysozyme (Lyz), which is abundant in the lacrimal fluid. Since chitosan (CS) can entrap more of the drug and also undergo hydrolysis in the presence of Lyz, we have employed CS for the composite preparation. The CL fabrication was performed by free radical copolymerization of poly(2-hydroxyethyl methacrylate) (pHEMA) in the presence of the drug-loaded nanocomposite with UV-curing initiators using the pre-drug loading strategy. The surface morphological, optical and mechanical investigations confirmed the presence of the drugs, ≥80% transparency, the adequate flexibility and biocompatibility of both the CLs. The in vitro release experiments showed the release of 95.86% LP from LP-CL, and 83.87% LP and 86.70% TM from LP-TM-CL in the presence of 1.5 mg mL-1 of Lyz in 72 h. In vitro biocompatibility assay against human corneal epithelial (HCE) cells and ex vivo experiments on HET-CAM confirmed that the fabricated LP-CL and LP-TM-CL are well tolerated. Moreover, in vivo safety evaluations of CLs on New Zealand white rabbit eyes suggest no sign of irritation to the ocular tissues within 72 h of observation. Hence, the study suggests that the 'single' and 'dual' drug-loaded CLs could open a new avenue to manage glaucoma by maintaining mean diurnal IOP.

Combination of self-assembling system and N,O-carboxymethyl chitosan improves ocular residence of anti-glaucoma drug.

Glaucoma is known to be one of the principal causes of vision loss due to elevated intraocular pressure. Currently, latanoprost eye drops is used as first-line treatment for glaucoma; however, it possesses low bioavailability due to rapid precorneal clearance. A novel delivery system with a mucoadhesive property could overcome this problem. Therefore, we attempt to develop a combination of self-assembling latanoprost nanomicelles (Latcel) and a mucoadhesive polymer (N,O-carboxymethyl chitosan: N,O-CMC) to improve the corneal residence time. Latcel was developed using Poloxamer-407 by thin film hydration method, followed by the addition of N,O-CMC using simple solvation to obtain Latcel-CMC and characterized using various physicochemical characterization techniques. The particle size of Latcel-CMC was 94.07 ± 2.48 nm and a zeta potential of -16.03 ± 0.66 mV, with a sustained release for 24h whereas marketed latanoprost drops released 90 % of the drug within 1h. In vitro cytotoxicity studies, HET-CAM, and in vivo Draize test showed the biocompatibility of Latcel-CMC. Cellular uptake studies performed using fluorescein isothiocyanate (FITC) loaded nanomicelles in human corneal epithelial cells indicates the increased cellular uptake as compare to plain FITC solution. In vivo ocular residence time was evaluated in Wistar rats using Indocyanine green (ICG) loaded nanomicelles by an in vivo imaging system (IVIS), indicating Latcel-CMC (8h) has better residence time than plain ICG solution (2h). The Latcel-CMC showed improved corneal residence time and sustained release of latanoprost due to increased mucoadhesion. Thus, the developed N,O-Carboxymethyl chitosan based nanomicelles eye drop could be a better strategy than conventional eye drops for topical delivery of latanoprost to treat glaucoma.

A clinical pilot study for the detection of sphingomyelinase in leptospirosis patient's urine at tertiary care hospital.

Purpose: Leptospirosis is a perplexing mystification for many clinicians. Clinically often underdiagnosed due to lack of a rapid, sensitive, and specific diagnostic test. Currently available diagnostic tests have their own limitations; therefore, monitoring biomarkers that contribute an essential role in pathogenesis is crucial. Herein, a pilot study was conducted to detect the presence of sphingomyelinase in urine of leptospirosis patients. Methods: Blood and urine samples were collected from 140 patients having febrile illness. Samples were analyzed through culturing, dark-field microscopy, detecting anti-leptospiral antibodies by MAT, IgM ELISA, Leptocheck-WB and screening for sphingomyelinase using a sphingomyelinase assay kit. Results: Out of 140 febrile illness patients, 22.14 % were tested leptospirosis, 33.57 % were dengue, 25 % scrub typhus, 18.57 % malaria and 0.71 % co-infection (dengue-leptospirosis). MAT seropositivity of 19.28 % (27/140) was confirmed with the highest agglutinant determined against serovar Icterohaemorrhagiae RGA followed by Autumnalis, Australis, and Pyrogens. IgM ELISA and Leptocheck-WB positivity was 16.42 % and 13.57 % respectively. Whereas culture and dark-field microscopy showed a sensitivity of 4.28 % and 2.1 %, respectively. Out of 31 confirmed cases of leptospirosis, sphingomyelinase was detected in the urine of 25 (80.64 %) patients, MAT positivity was seen in 87.09 % and culture positivity was seen in 12.90 % of cases. Conclusion: Detection of sphingomyelinase in the urine of a leptospirosis patient and its absence in other febrile illnesses like dengue, malaria and scrub typhus establish evidence of secretion of sphingomyelinase in urine during leptospiral infection. Hence, sphingomyelinase could be used as a potential diagnostic biomarker to detect leptospirosis in a non-invasive way.

Development of mucoadhesive Timolol loaded chitosan-nanocomposite to treat glaucoma.

Timolol Maleate is an aqueous soluble ß-blocker antiglaucoma drug used to suppress intraocular pressure. Several commercially available ocular formulations are not effective in delivering to the target site due to their water-soluble property and low mucoadhesiveness. Hence, there is a requirement for a highly mucoadhesive drug-loaded nanocomposite to suppress intraocular pressure with enhanced bioavailability. Herein, we have prepared a mucoadhesive Timolol-loaded graphene quantum dot-chitosan-nanocomposite to treat glaucoma in response to lysozyme, secreted in the tear fluid. The as-prepared nanocomposite has been characterized through high resolution-transmission electron microscopic, X-ray photoelectron spectroscopic, X-ray diffraction, and Fourier transform infrared spectral studies. The nanocomposite showed 93.74 % encapsulation efficiency with a loading capacity of 7.73 %. Further, 89.26 %, 95.62 %, and 99.29 % of drug release were observed from the nanocomposite in the presence of 1, 1.5, and 2 mg/mL of lysozyme. The mucoadhesion property has been confirmed by the increment in the particle size, fluorescence spectral variations, and Fourier transform infrared spectroscopic studies in the presence of mucin nanoparticles of size 291 nm. Interestingly, mucoadhesion has been demonstrated by pointing to the quenching in the luminescence of mucin. Further, in vitro biocompatibility assay on human corneal epithelial cells showed ≥80 % cell viability. Hence, this study offers the utilization of naturally secreting enzymes for drug delivery applications instead of uncontrolled pH and temperature-triggered releases.

Direct Electron Transfer of Glucose Oxidase on Pre-Anodized Paper/Carbon Electrodes Modified through Zero-Length Cross-Linkers for Glucose Biosensors.

A disposable paper-based glucose biosensor with direct electron transfer (DET) of glucose oxidase (GOX) was developed through simple covalent immobilization of GOX on a carbon electrode surface using zero-length cross-linkers. This glucose biosensor exhibited a high electron transfer rate (ks, 3.363 s-1) as well as good affinity (km, 0.03 mM) for GOX while keeping innate enzymatic activities. Furthermore, the DET-based glucose detection was accomplished by employing both square wave voltammetry and chronoamperometric techniques, and it achieved a glucose detection range from 5.4 mg/dL to 900 mg/dL, which is wider than most commercially available glucometers. This low-cost DET glucose biosensor showed remarkable selectivity, and the use of the negative operating potential avoided interference from other common electroactive compounds. It has great potential to monitor different stages of diabetes from hypoglycemic to hyperglycemic states, especially for self-monitoring of blood glucose.

Ni/Ni(OH)2-rGO nanocomposites sensor for the detection of long forgotten mycotoxin, xanthomegnin.

Xanthomegnin, a known fungal toxin, secondary metabolite, and pigment diffuses from the dermatophytes has gained attention as local virulence factor because of the mutagenicity, toxicity, cytocidal, and immunosuppressive properties. Not only as a dermatophyte in skin related disorders, the production of xanthomegnin is implicated as a powerful diagnostic marker in patients suffering from ocular mycoses. Incidentally also attributed to death in livestock's majorly by exposing themselves to food-borne fungi like Aspergillus and Penicillium. The production of xanthomegnin in dermetophytic species Trichophyton rubrum, found commonly in infected skin and nails. In this study nickel/nickel hydroxide nanoparticles decorated reduced graphene oxide (Ni/Ni(OH)2-rGO) modified glassy carbon electrode has been successfully used for non-enzymatic detection of xanthomegnin. The Ni/Ni(OH)2-rGO composites were synthesized through a simple microwave assisted technique with less harmful reducing agent. The UV-visible spectroscopy (UV-vis), X-ray photoelectron spectroscopy (XPS), Scanning electron microscopy - energy dispersive X-ray spectroscopy (SEM-EDS), and electrochemical investigations demonstrated the robust formation of the sensor. The sensor exhibited improved electrochemical properties with enhanced electrochemical active area and excellent electrochemical behavior towards xanthomegnin detection with a limit of detection of 0.12 µM. The selectivity, stability, and analytical recovery studies proved the potential use of the sensor for the detection of xanthomegnin in real samples. Further, the sensor successfully detected xanthomegnin produced by the Trichophyton rubrum, the most common superficial fungus, accounting for at least 60% of all superficial fungal infections in humans. Validation studies showed satisfiable and quantifiable amount of xanthomegnin in comparison with common bench mark UV-Vis studies meant for fungal mycotoxin detection.

Leptospiral sphingomyelinase Sph2 as a potential biomarker for diagnosis of leptospirosis.

Leptospirosis is an underestimated infectious tropical disease caused by the spirochetes belonging to the genus Leptospira. Leptospirosis is grossly underdiagnosed due to its myriad symptoms, varying from mild febrile illness to severe haemorrhage. Laboratory tests for leptospirosis is an extremely important and potent way for disease diagnosis, as the clinical manifestations are very similar to other febrile diseases. Currently available diagnostic techniques are time-consuming, require expertise and sophisticated instruments, and cannot identify the disease at an early phase of infection. Early diagnosis of leptospirosis is the need of the hour while considering the severe complications after the infection and the rate of mortality after misdiagnosis. Secretion of Leptospira-specific sphingomyelinases in leptospirosis patient's urine within a few days of the onset of infection is quite common and is a virulence factor present only in pathogenic Leptospira species. Herein, the structural and functional importance of leptospiral sphingomyelinase Sph2 in leptospirosis pathogenesis, as well as the potential of screening urinary Sph2 for diagnosis and the scope for developing a rapid and easily affordable point-of-care test for urinary leptospiral sphingomyelinase Sph2 as an alternative to current diagnostic methods are discussed.

Novel chitosan - graphene quantum dots composite for therapeutic delivery and tracking through enzymatic stimuli response.

The designing of highly efficient and biocompatible nanocomposites with multifunctional delivery and tracking characteristics is noteworthy for clinical and therapeutic applications. Herein, we report the proof-of-concept for the delivery of anti-glaucoma drug, latanoprost (LP) under an enzymatic stimulus, lysozyme (Lyz) with novel chitosan (CS) - graphene quantum dots (GQD) nanocomposite via reverse switching photoluminescence (PL) phenomenon. The LP caged CS-GQDs nanocomposite was well characterized through extensive spectral, morphological, band-gap, particle size, and zeta potential studies along with cytotoxicity assays. The regaining of PL not only confirmed LP delivery, but also facilitated intercellular tracking through in vitro bio-imaging against human corneal epithelial (HCE) cells. The AO/EB staining and biocompatibility assays further proved excellent cell viability of >80%. The successfully delivered LP protected HCE cells from oxidative injury induced by 800 µM hydrogen peroxide (H2O2). These findings justify further utility of novel CS-GQDs caged drug nanocomposite for preclinical investigations.

Evaluation of anti-LipL32 carbon nanotube immunofluorescence probe (carbo-lip) and comparison with MAT, IgM ELISA, IgM spot test and culture for early detection of leptospirosis at local hospital.

Leptospirosis is an emerging public health problem affecting people mainly from tropical and subtropical regions. Therefore, there is a need for rapid and sensitive tests for proper and prompt treatment. Recently we have demonstrated Carbo-Lip probe, which was fabricated through immuno recognition method with fluorescent dye functionalized LipL32 monoclonal antibodies, secondary antibody and Leptospira for rapid and accurate diagnosis. In an effort to validate Carbo-Lip, we collected clinical samples from a cohort of 104, consisting of 26 positive, 40 negative and 38 unconfirmed cases of Leptospirosis. Subsequently, the test was also compared and validated with the gold standard method microscopic agglutination test (MAT), IgM ELISA, IgM spot test, and culture. Carbo-Lip exhibited a sensitivity of 75% with specificity of 92.3% for Leptospirosis in comparison with MAT. The fabricated Carbo-Lip sensor could be used as a potential diagnostic tool for early detection of Leptospirosis in patients from endemic areas.

Paper-based field-effect transistor sensors.

The present scenario in the world largely demands affordable, fast, recyclable, and flexible electronic devices for bio sensing. Varieties of paper-based devices such as microfluidics paper electrodes, paper diodes, and paper-based transistors etc. have been developed and validated. Most of the fabrication techniques published so far have focused on economic, environment-friendly straightforward methods to develop paper-based field-effect transistors (PFET) sensors, additionally, explored their applications. The synthetic-free, mechanically flexible, biocompatible, and signal amplification capability render PFET based sensors for wearable device makers. Modified organic/inorganic PFETs identify target analytes based on the electrical signal and endow them as perfect transducers. In the field of PFET bio sensing technology, numerous challenges are needed to be discussed to proceed forward in biomedical and other analytical applications. Realizing biologically or chemically modified PFET having an exceptional signal to noise ratio, specificity, with rapid detection ability is challenging. This review recapitulates the fabrication techniques, performances of various PFET sensors, and summarizes the report by concluding remarks including the limitations of the existing PFET based sensors and the future holds in regards to the sustainable nature of PFET.

Towards CRISPR powered electrochemical sensing for smart diagnostics.

Even though global health has been steadily improved, the global disease burden associated with communicable and non-communicable diseases extensively increased healthcare expenditure. The present COVID-19 pandemic scenario has again ascertained the importance of clinical diagnostics as a basis to make life-saving decisions. In this context, there is a need for developing next-generation integrated smart real-time responsive biosensors with high selectivity and sensitivity. The emergence of clustered regularly interspaced short palindromic repeats (CRISPR)/Cas biosensing systems has shown remarkable potential for developing next-generation biosensors. CRISPR/Cas integrated electrochemical biosensors (E-CRISPR) stands out with excellent properties. In this opinionated review, we illustrate the rapidly evolving applications for E-CRISPR-integrated detection systems towards biosensing and the future scope associated with E-CRISPR based diagnostics.

Why chitosan could be apt candidate for glaucoma drug delivery - An overview.

Most of the people in the world are affected by glaucoma, which leads to irreversible blindness. Several patient friendly treatments are available, nevertheless medications lack an easy and efficient way of sustained delivery. To make the delivery with enhanced bioavailability, biodegradable and non-biodegradable polymers-based drug carriers are explored. However, ocular drug delivery issues have not been resolved yet due to less adhesiveness, poor penetration ability, pH, and temperature dependent burst releases. Chitosan is found to be effective for ocular drug delivery due to excellent physio-chemical properties in terms of overcoming the existing issues. In this review, we aim to highlight why it has been chosen and the holy grail for ocular drug delivery. Besides, we have comprehensively reviewed recent patents on chitosan as a platform for ocular drug delivery and future perspectives on factors, lacunae and challenges that need to be addressed for better ocular delivery methods for glaucoma management.

A new method to amplify colorimetric signals of paper-based nanobiosensors for simple and sensitive pancreatic cancer biomarker detection.

A low-cost, sensitive, and disposable paper-based immunosensor for instrument-free colorimetric detection of pancreatic cancer biomarker PEAK1 was reported for the first time by capitalizing the catalytic properties of gold nanoparticles in colour dye degradation. This simple signal amplification method enhances the detection sensitivity by about 10 fold.

Synthesis, characterization, and electrochemical applications of carbon nanoparticles derived from castor oil soot.

A simple procedure for the modification of carbon nanoparticles (CNPs) from castor oil soot using acid treatment was described herein. Characterization studies revealed the presence of edge plane sites and surface carbon-oxygen functionalities at the surface of the CNP material. Voltammetric studies revealed the increased electrochemical activity of the CNP-modified electrode toward various biologically important molecules, including dopamine, uric acid, dihydronicotinamide adenine dinucleotide, tyrosine, and serotonin, relative to those obtained using the unmodified electrode. The improved electro-oxidation potentials for these compounds-and, thereby, the enhanced sensitivity of related sensors-was due directly to the presence of surface C(δ+)O(δ-) functional groups and the greater number of edge plane sites developed after acid treatment of the soot sample.