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BACKGROUND: This study evaluated physicians' and patients' beliefs about biosimilars in Hong Kong, India, Pakistan, Singapore, Taiwan, and Thailand. RESEARCH DESIGN AND METHODS: An online survey administered to physicians (dermatologists, n = 119; gastroenterologists, n = 148; rheumatologists, n = 161) between 22 October 2021 and 7 January 2022, and patients (n = 90) with rheumatic or inflammatory bowel disease between 25 October 2021 and 12 April 2022. RESULTS: Most (68%) physicians reported having a strong knowledge about biosimilars, yet 49% indicated that biosimilars are readily available to them. Physicians cited potential cost savings (81%) as the main benefit of biosimilars, and cost/coverage support (36%), patient support (25%), and increasing biosimilar awareness/education (24%) as main strategies for improving usage. Few (21%) patients reported having a strong knowledge about biosimilars. Patients cited offering alternatives in case of drug shortages (77%) as the main benefit of biosimilars, and cost/coverage support (53%), increasing awareness of product profile (22%), and providing biosimilars with a good efficacy profile/effective product (19%) as main strategies for improving usage. CONCLUSION: Programs focused on cost/coverage support, patient support, and biosimilar awareness could improve acceptance of biosimilars for chronic immune-mediated inflammatory diseases in Asian countries, thereby increasing patient access to essential biologic therapies.

Biologic medicines come from living organisms such as viruses, bacteria, and animal or plant cells. Biosimilars are approved biologic medicines that are highly similar in structure to original biologic medicines. This study was done to understand what people with diseases that cause long-lasting inflammation of the joints, intestines, or skin (inflammatory diseases for short) and their doctors think about biosimilars. Researchers focused on people and doctors living in six Asian countries (Hong Kong, India, Pakistan, Singapore, Taiwan, and Thailand) with economies and health systems that are advanced or becoming more advanced. Researchers used an online survey to ask people with inflammatory diseases (or people responders) (90 in total) and their doctors (428 in total) what they think about biosimilars. Most doctors (68%) and some people responders (21%) said they had a good understanding of biosimilars. Doctors thought the main benefit of using biosimilars was the possibility of achieving cost savings. People responders thought the main benefit was the possibility of having additional treatment options in case of shortages of the original biologic medicine. Doctors said they sometimes cannot prescribe biosimilars because the cost may still be too high for some patients or because they worry about the quality of biosimilars and how well they work. Programs that increase awareness of biosimilars and that provide financial and other support for biosimilars could improve biosimilar use as treatment for inflammatory diseases in Asian countries, giving more patients access to important biologic medicines in this region.

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Chlorpyrifos (CPS) is widely found in food and water sources due to agricultural use, posing health and environmental risks. Therefore, this work introduces a fluorescent sensor design of silver nanoparticle-embedded nano zirconium-based metal-organic frameworks (UiO-66-NH2@AgNPs) for accurate examination of CPS. Briefly, UiO-66-NH2 was synthesized hydrothermally, exhibiting weak luminescence owed to ligand-to-metal charge transfer (LMCT). Here, it limits its direct utility in fluorescence-based detection. To address this limitation, silver nanoparticles (AgNPs) were introduced into UiO-66-NH2, enhancing fluorescence via the metal-enhanced fluorescence (MEF) effect. Briefly, a comprehensive spectral analysis such as XPS, SEM, TEM, PXRD, etc., was performed to validate the synthesis of UiO-66-NH2@AgNPs. Subsequent evaluation revealed that CPS effectively quenched the luminescence intensity of UiO-66-NH2@AgNPs through a static quenching mechanism. The fluorescence intensity exhibited good linearity with CPS concentration in the span of 10 to 1,000 ng/mL, with a recognition limit of 191.5 ng/mL(S/N = 3). The interaction involved Ag-S bond formation and electrostatic interactions, reducing fluorescence intensity. The method was confirmed through successful CPS detection in fruit samples. The UiO-66-NH2@AgNPs nanoprobe offers a simple, sensitive, and accurate platform for CPS sensing, with potential for future use in detecting CPS in fruits and vegetables.

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In this study, we developed a new fluorescence "on-off-on" sensor utilizing water-soluble cobalt/zinc-nitrogen co-doped graphene quantum dots (Co/Zn-N-GQDs) to recognize quinalphos pesticide in vegetable and fruit samples. Primarily, the synthesis method employed a one-pot hydrothermal approach, using betel leaves as a natural precursor and cobalt ("Co"), zinc ("Zn"), and urea ("N") as dopant sources. The Co/Zn-N-GQDs probes underwent comprehensive analytical characterization. The Co/Zn-N-GQDs were synthesized with a remarkable luminescence yield of 31.49%, exhibiting excitation at 320 nm and emission peak at 393 nm. Interestingly, the luminescence of Co/Zn-N-GQDs was selectively "Turned Off" by Cu2+ via a static quenching setup. Remarkably, quenched fluorescence was surprisingly reactivated upon adding quinalphos to the quench setup, indicating a direct correlation between luminescence reactivation and quinalphos concentration. Briefly, this phenomenon is ascribed to the functional groups in quinalphos, such as quinoxalinyl and phosphorothioate, which chelate with Cu2+ ions, disrupting the nonfluorescent Cu2+-Co/Zn-N-GQDs complex. The design sensor demonstrated a limit of detection (LOD) of 0.11 µM and a broad linear span of 0.5 to 200 µM. In conclusion, Cu2+-Co/Zn-N-GQDs sensor showed immediate applicability, stability, and reproducibility, making it highly effective for quinalphos sensing in various samples.

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Aim: The study was aimed to formulate and evaluate apremilast-loaded zinc oxide-mesoporous silica nanoparticles for treatment of psoriasis. Materials & methods: Mesoporous silica nanoparticles were prepared by using sol-gel method and evaluated for particle size, in vitro drug release, in vitro cytotoxicity study and in vivo pharmacodynamic study. Results: The synthesized mesoporous silica nanoparticles showed particle size of 319.9 ± 3.9 nm, with 24 ± 0.217% of loading capacity. In vitro cytotoxicity study on A-431 cell line showed increased anti-psoriatic activity of apremilast-loaded zinc oxide-mesoporous silica nanoparticles. In vivo pharmacodynamic study and histological studies showed improved efficacy of drug in imiquimod-induced psoriasis mice model. Conclusion: The apremilast-loaded zinc oxide-mesoporous silica nanoparticles showed improved therapeutic efficacy, suggesting that they are promising approach for topical treatment of psoriasis.

[Box: see text].

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AIM: Atorvastatin (ATO) loaded chitosan-based polyelectrolyte complex nanoparticles (PECN) incorporated transdermal patch was developed to enhance its skin permeability and bioavailability. METHODOLOGY: The ATO loaded PECN were prepared by ionic gelation method and optimized by Box-Behnken design. The optimized batches were evaluated for physicochemical characteristics, in vitro, ex vivo, cell line and stability studies. The optimized ATO-PECN were incorporated into transdermal patches by solvent evaporation method and evaluated for their physicochemical properties, ex vivo skin permeation, in vivo pharmacokinetics and stability study. RESULTS: The optimized batch of ATO-PECN had average size of 219.2 ± 5.98 nm with 82.68 ± 2.63 % entrapment and 25.41 ± 3.29 mV zeta potential. ATO-PECN showed sustained drug release and higher skin permeation. The cell line study showed that ATO-PECN increased the cell permeability of ATO as compared to ATO suspension. ATO-PECN loaded transdermal patch showed higher skin permeation. The in vivo pharmacokinetic study revealed that the ATO-PECN transdermal patch showed significant (p < 0.05) increase in pharmacokinetic parameters as compared to marketed oral tablet, confirming enhancement in bioavailability of ATO. CONCLUSIONS: The results of the present work concluded that the ATO-PECN loaded transdermal patch is a promising novel drug delivery system for poorly bioavailable drugs.

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A spectrofluorimetric method using fluorescent carbon dots (CDs) was developed for the selective detection of azelnidipine (AZEL) pharmaceutical in the presence of other drugs. In this study, N-doped CDs (N-CDs) were synthesized through a single-step hydrothermal process, using citric acid and urea as precursor materials. The prepared N-CDs showed a highly intense blue fluorescence emission at 447 nm, with a photoluminescence quantum yield of ~21.15% and a fluorescence lifetime of 0.47 ns. The N-CDs showed selective fluorescence quenching in the presence of all three antihypertensive drugs, which was used as a successful detection platform for the analysis of AZEL. The photophysical properties, UV-vis light absorbance, fluorescence emission, and lifetime measurements support the interaction between N-CDs and AZEL, leading to fluorescence quenching of N-CDs as a result of ground-state complex formation followed by a static fluorescence quenching phenomenon. The detection platform showed linearity in the range 10-200 µg/ml (R2 = 0.9837). The developed method was effectively utilized for the quantitative analysis of AZEL in commercially available pharmaceutical tablets, yielding results that closely align with those obtained from the standard method (UV spectroscopy). With a score of 0.76 on the 'Analytical GREEnness (AGREE)' scale, the developed analytical method, incorporating 12 distinct green analytical chemistry components, stands out as an important technique for estimating AZEL.

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Integrating enzymes and nanozymes in various applications is a topic of significant interest. The researchers have explored the encapsulation of enzymes using diverse nanostructures to create nanomaterial-enzyme hybrids. These nanomaterials introduce unique properties that contribute to the additional activity along with the stabilization of enzymes in immobilized form, enabling a cascade of second-order reactions. This review centers on dual-activity nanozymes, providing insights into their applications in biosensors and biocatalysis. These applications leverage the enhanced catalytic activity and stability offered by dual-activity nanozymes. These nanozymes find promising applications in fields like bioremediation, offering eco-friendly solutions for mitigating environmental pollution while showing potential in medical diagnostics. The review delves into various techniques for creating enzyme-nanozyme hybrid catalysts, including adsorption, encapsulation, and incorporation methods. The review also addresses the challenges that must be overcome, such as overlapping catalytic surfaces and disparities in reaction rates in multi-enzyme cascade reactions. It concludes by presenting strategies to tackle these issues and offers insights into the field's promising future, suggesting that machine learning may drive further advancements in enzyme-nanozyme integration. This comprehensive exploration illuminates the present and charts a promising course for future innovations in the seamless integration of enzymes and nanozymes, heralding a new era of catalytic possibilities.

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Biodiesel production through the synthesis of Datura stramonium L. oil is studied to explore the most efficient approaches to suggest an alternate feedstock for biodiesel production. The main objective of this work is to optimize the process variables of biodiesel synthesis by using some statistical approach (Taguchi method, grey relational analysis (GRA), and response surface methodology (RSM) analyzing three parameters, i.e., alcohol-to-oil molar ratio, catalyst (NaOH) concentration, and process temperature for achieving maximum biodiesel derived from Datura stramonium L. oil. The transesterification process is applied by using an ultrasonic-assisted technique. Grey relational analysis (GRA) was successfully applied with the Taguchi method resulting in the optimum combination of A2B1C1. Based on the findings, the best operating conditions for transesterifying are attained with the RSM approach consisting of a 5.697:1 molar ratio (level 2), 0.3 (wt.%) NaOH concentration (level 1), and 70 °C process temperature (level 1). With a value of 87.02%, these ideal operating conditions produce the maximum yield as compared to grey relational analysis (GRA) yields 83.99%. The obtained results have been verified through the characterization of oil and biodiesel as well. Also, the fuel qualities of DSL biodiesel were identified and assessed. DSL oil was found 137.6 degrees of unsaturation during fatty acid profile analysis. DSL biodiesel was found the best kinematic viscosity (4.2 mm2/s) and acid value (0.49) when compared to Karanja and palm biodiesel. D. stramonium L. was recognized as a suitable species for biodiesel feedstock according to the findings.

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Despite the high medicinal value of tiopronin, there are substantial adverse effects such as yellow skin, yellow eyes, muscle aches, etc. Therefore, there is a huge necessity to identify tiopronin using advanced sensors in provided samples. Recently, the preference for graphene quantum dots (GQDs) and inorganic nanomaterial-based fluorescent sensors for the detection of pharmaceuticals has been extensively documented due to their plentiful advantages. Therefore, in this work, the cobalt-doped GQDs decorated vanadium pentoxide nanosheet-based fluorescence switch 'Off-On' sensor (Co-GQDs@V2O5-NS) was designed for highly sensitive and selective detection of tiopronin. Briefly, the green synthesis of highly fluorescent Co-GQDs was carried out using a hydrothermal method. Meanwhile, the synthesis of V2O5-NS was synthesized using the liquid exfoliation method. The synthesis of Co-GQDs@V2O5-NS was accomplished wherein Co-GQDs adsorbed on the surface of V2O5-NS that offered the quenching of fluorescence of Co-GQDs. Afterward, the addition of tiopronin into the quenched probe disclosed the proportional recovery of fluorescence of Co-GQDs. Here, the addition of tiopronin provides the decomposition of V2O5-NS and conversion into the V4+ that aids in releasing the quenched fluorescence of Co-GQDs. The limit of detection and linearity range for tiopronin was found to be 1.43 ng/mL and 10-700 ng/mL, respectively. Moreover, it demonstrated high selectivity, good stability at experimental conditions, and practicality in analyzing tiopronin in spiked sample analysis. Hence, the designed Co-GQDs@V2O5-NS nanosized sensor enables high sensitivity, selectivity, simplicity, label-free, and eco-friendly tiopronin recognition. In the future, the utility of Co-GQDs@V2O5-NS can open a new door for sensing tiopronin in provided samples.

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Background: US FDA defines: dietary supplements is a product that intended to supplement a person's diet, it's generally consist of at least one or more of the following dietary ingredients, vitamin, minerals, a herb or other botanical and amino acid by increasing the daily consumptions of an extract metabolite concentration, constitute or combinations of these medication. Excessive and inappropriate use of medicines has been recognised as a public health problem resulting in increased likelihood of adverse drug event, drug interaction, and inappropriate drug prescribing and increased cost. Material and Methods: This was the cross-sectional study conducted in year 2022 at Pimpri Chinchwad (Pune). The total 250 questionaires are distributed and from that 226 response were received. Target population consist of community pharmacists working in the drug store in this area (n=226). Results: Data was represented in three domains of study i. e. awareness, knowledge and attitude. Correlation coefficient using Pearson's method were determined to evaluate strength of correlation between awareness-knowledge, Knowledge-attitude and awareness-attitude. Correlation coefficient were calculated by comparing most relevant and equal number of questions. Conclusion: The study demonstrated positive attitude among surveyed community pharmacists in Pune, India. There is lacuna in accurate and adequate awareness, knowledge and attitude of vitamin deficiency, efficiency, recommended daily allowance (RDA), toxicity and interactions among pharmacist as one of the stakeholders of healthcare in India. Few of the remedies viz. framing of guidelines, inclusion in formal education syllabus, continuous education, updation exams etc. may be of use.

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Clozapine, which is widely used to treat schizophrenia, shows low bioavailability due to poor solubility and high first-pass metabolism. The study aimed to design clozapine-loaded carbon dots (CDs) to enhance availability of the clozapine to the brain via intranasal pathway. The CDs were synthesized by pyrolysis of citric acid and urea at 200 °C by hydrothermal technique and characterized by photoluminescence, transmission electron microscopy (TEM), X-ray Photoelectron Spectrometer (XPS), and Fourier transform infrared spectrum (FTIR). The optimized clozapine-loaded CDs (CLZ-CDs-1:3-200) showed a quasi-spherical shape (9-12 nm) with stable blue fluorescence. The CDs showed high drug solubilization capacity (1.5 mg drug in 1 mg/ml CDs) with strong electrostatic interaction with clozapine (drug loading efficiency = 94.74%). The ex vivo release study performed using nasal goat mucosa showed sustained release of clozapine (43.89%) from CLZ-CDs-1:3-200 for 30 h. The ciliotoxicity study (histopathology) confirmed no toxicity to the nasal mucosal tissues using CDs. In the rat model (in vivo pharmacokinetic study), when CDs were administrated by the intranasal route, a significantly higher concentration of clozapine in the brain tissue (Cmax = 58.07 ± 5.36 µg/g and AUCt (µg/h*g) = 105.76 ± 12.31) was noted within a short time (tmax = 1 h) compared to clozapine suspension administered by intravenous route (Cmax = 20.99 ± 3.91 µg/g, AUC t (µg/h*g) = 56.89 ± 12.31, and tmax = 4 h). The high value of drug targeting efficiency (DTE, 486%) index and direct transport percentage (DTP, 58%) indicates the direct entry of clozapine-CDs in the brain via the olfactory route. In conclusion, designed CDs demonstrated a promising dosage form for targeted nose-to-brain delivery of clozapine for the effective treatment of schizophrenia.

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We designed a highly sensitive fluorescent sensor for the early detection of sarcosine, a potential biomarker for prostate cancer. This sensor was based on surface-cobalt-doped fluorescent carbon quantum dots (Co-CD) using a FRET-based photoluminescent sensing platform. Blue luminescent carbon quantum dots (CQD) were synthesised through a hydrothermal approach, utilizing Delonix regia tree pod shells. Cobalt was employed to functionalize the CQD, enhancing the quantum-entrapped effects and minimizing surface flaws. To optimize Co-CD preparation, we employed a Box-Behnken design (BBD), and response surface methodology (RSM) based on single-factor experiments. The Co-CD was then used as a fluorescent probe for selective Cu2+ detection, with Cu2+ quenching Co-CD fluorescence through an energy transfer process, referred to as 'turn-off'. When sarcosine was introduced, the fluorescence intensity of Co-CD was restored, creating a 'turn-on' response. The sensor exhibited a Cu2+ detection limit (LOD) of 2.4 µM with a linear range of 0 µM to 10 µM. The sarcosine detection in phosphate buffer saline (PBS, pH 7.4) resulted in an LOD of 1.54 µM and a linear range of 0 to 10 µM. Importantly, the sensor demonstrated its suitability for clinical analysis by detecting sarcosine in human urine. In summary, our rapid and highly sensitive sensor offers a novel approach for the detection of sarcosine in real samples, facilitating early prostate cancer diagnosis.Communicated by Ramaswamy H. Sarma.

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Hepatocellular carcinoma (HPTC) currently ranks as the third leading cause of cancer-related mortality, necessitating an advanced formulation strategy. Recently, lactoferrin (Lf) has been utilized as a specific targeting ligand in HPTC due to its high specificity towards the asialoglycoprotein receptor expressed in cancer cells. Therefore, we present the fabrication of an Lf-decorated carboxymethyl dextran-encased chitosan-coated europium metal-organic framework-based nanobioconjugate (Lf-CMD-CS-CUR@Eu-MOF) for targeted curcumin (CUR) delivery. Briefly, CUR was loaded into Eu-MOF, followed by coating cationic 'CS' on the CUR@Eu-MOF surface. Simultaneously, Lf-decorated CMD was prepared via an esterification reaction. Subsequently, Lf-CMD-CS-CUR@Eu-MOF was synthesized using the Maillard reaction. Various spectral characterizations, drug entrapment, drug content, in vitro drug release, biocompatibility and cell cytotoxicity studies were performed. It exhibited an entrapment efficiency of 88.87 ± 2.1 %, a drug content of 3.45 ± 0.98 %, and a drug loading rate of 34.85 ± 0.6 mg/g. Furthermore, the Lf-CMD-CS-CUR@Eu-MOF exhibits excellent biocompatibility with normal cells. The in vitro dissolution study confirmed a release of 78.12 % of 'CUR' in pH 5.8 phosphate buffer (over 120 h), attributed to the controlled release rate by the 'CS' coating on the surface of CUR@Eu-MOF. The BEL-7402 cell line showed concentration-dependent toxicity of nanobioconjugate to cancerous cells. Therefore, when 'Lf' is surface-decorated onto an appropriate polymeric material, it gains the capability to function as a carrier for transporting 'CUR' to the precise target site within HPTC. In conclusion, Lf-CMD incorporated CS-coated Eu-MOF can provide a promising approach for targeted drug delivery in HPTC management.

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The increased mortality rates associated with colorectal cancer highlight the pressing need for improving treatment approaches. While capsaicin (CAP) has shown promising anticancer activity, its efficacy is hampered due to low solubility, rapid metabolism, suboptimal bioavailability, and a short half-life. Therefore, this study aimed to prepare a lactoferrin-functionalized carboxymethyl dextran-coated egg albumin nanoconjugate (LF-CMD@CAP-EGA-NCs) for the targeted CAP delivery to enhance its potential for colorectal cancer therapy. Briefly, LF-CMD was synthesized through an esterification reaction involving LF as a receptor and CMD as a shell. Concurrently, CAP was incorporated into an EGA carrier using gelation and hydrophobic interactions. The subsequent production of LF-CMD@CAP-EGA-NCs was achieved through the Maillard reaction. Spectral characterizations confirmed the successful synthesis of smooth and spherical-shaped LF-CMD@CAP-EGA-NCs using LF-CMD and EGA-CAP nanoparticles, with high entrapment efficiency and satisfactory drug content. Furthermore, LF-CMD@CAP-EGA-NCs demonstrated a sustained release of CAP (76.52 ± 1.01 % in 24 h, R2 = 0.9966) in pH 5.8 buffer with anomalous transport (n = 0.68) owing to the shell of the CMD and EGA matrix. The nanoconjugate exhibited enhanced cytotoxicity in HCT116 and LoVo cell lines, which is attributed to the overexpression of LF receptors in colorectal HCT116 cells. Additionally, LF-CMD@CAP-EGA-NCs demonstrated excellent biocompatibility, as observed in the FHC-CRL-1831 cell line. In conclusion, LF-CMD@CAP-EGA-NCs can be considered as a promising approach for targeted delivery of CAP and other anticancer agents in colorectal cancer treatment.

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Presently, there is a necessity to design novel methods because of quercetin's significant biological relevance. Therefore, we developed the rose petal-derived graphene quantum dots embedded zinc metal organic frameworks (RP-GQDs@Zn-MOFs) based fluorescence "On-Off-On" nanoprobe for quercetin sensing. Initially, RP-GQDs were synthesized using rose petal waste and then subjected to embedding into Zn-MOFs. Herein, the addition of copper ions (Cu2+) results in fluorescence "Switch Off" whereas quercetin inclusion resulted in the formation of the quercetin-Cu2+ complex. It regains the RP-GQDs@Zn-MOFs quenched fluorescence termed as "Switch On" because of the static quenching mechanism. It demonstrated a wide concentration range and low detection limit of 100 ng/mL to 1400 ng/mL (R2=0.99) and 37.84 ng/mL, respectively. The selectivity study shows the high specificity for quercetin in presence of interfering compounds because of Cu2+ coordination between the carbonyl oxygen atom and the 3-OH group of quercetin. Moreover, the designed probe shows good stability, repeatability, and real-time analysis possibility.

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Cluster of differentiation (CD59), a cell surface glycoprotein, regulates the complement system to prevent immune damage. In cancer, altered CD59 expression allows tumors to evade immune surveillance, promote growth, and resist certain immunotherapies. Targeting CD59 could enhance cancer treatment strategies by boosting the immune response against tumors. Herein, we present a one-step synthesis of Polyethyleneimine (PEI) functionalized graphene quantum dots (Lf-GQDs) from weathered lemon leaf extract. The fabricated Lf-GQDs were successfully used for the quantitative detection of the cluster of CD59 antigen that is reported for its expression in different types of cancer. In this work, we utilized orientation-based attachment of CD59 antibody (Anti-CD59). Our findings reveal that, instead of using random serial addition of antigen or antibody, oriented conjugation saves accumulated concentration offering greater sensitivity and selectivity. The Anti-CD59@Lf-GQDs immunosensor was fabricated using the oriented conjugation of antibodies onto the Lf-GQDs surface. Besides, the fabricated immunosensor demonstrated detection of CD59 in the range of 0.01 to 40.0 ng mL-1 with a low detection limit of 5.3 pg mL-1. Besides, the cellular uptake potential of the synthesized Lf-GQDs was also performed in A549 cells using a bioimaging study. The present approach represents the optimal utilization of Anti-CD59 and CD59 antigen. This approach could afford a pathway for constructing oriented conjugation of antibodies on the nanomaterials-based immunosensor for different biomarkers detection.

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Conventional techniques for enzyme immobilization suffer from suboptimal activity recovery due to insufficient enzyme loading and inadequate stability. Furthermore, these techniques are time-consuming and involve multiple steps which limit the applicability of immobilized enzymes. In contrast, the use of microfluidic devices for enzyme immobilization has garnered significant attention due to its ability to precisely control immobilization parameters, resulting in highly active immobilized enzymes. This approach offers several advantages, including reduced time and energy consumption, enhanced mass-heat transfer, and improved control over the mixing process. It maintains the superior structural configuration in immobilized form which ultimately affects the overall efficiency. The present review article comprehensively explains the design, construction, and various methods employed for enzyme immobilization using microfluidic devices. The immobilized enzymes prepared using these techniques demonstrated excellent catalytic activity, remarkable stability, and outstanding recyclability. Moreover, they have found applications in diverse areas such as biosensors, biotransformation, and bioremediation. The review article also discusses potential future developments and foresees significant challenges associated with enzyme immobilization using microfluidics, along with potential remedies. The development of this advanced technology not only paves the way for novel and innovative approaches to enzyme immobilization but also allows for the straightforward scalability of microfluidic-based techniques from an industrial standpoint.

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The SN2 nucleophilic substitution reaction is a vital organic transformation used for drug and natural product synthesis. Nucleophiles like cyanide, oxygen, nitrogen, sulfur, or phosphorous replace halogens or sulfonyl esters, forming new bonds. Isocyanides exhibit unique C-centered lone pair σ and π* orbitals, enabling diverse radical and multicomponent reactions. Despite this, their nucleophilic potential in SN2 reactions remains unexplored. We have uncovered that isocyanides act as versatile nucleophiles in SN2 reactions with alkyl halides. This yields highly substituted secondary amides through in situ nitrilium ion hydrolysis introducing an alternative bond break compared to classical amide synthesis. This novel 3-component process accommodates various isocyanide and electrophile structures, functional groups, scalability, late-stage drug modifications, and complex compound synthesis. This reaction greatly expands chemical diversity, nearly doubling the classical amid coupling's chemical space. Notably, the isocyanide nucleophile presents an unconventional Umpolung amide carbanion synthon (R-NHC(-) = O), an alternative to classical amide couplings.

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Chitosan (CS) and sodium alginates (SA) have been revealed for the design of layer-by-layer (LbL) assembly to develop pharmaceutical dosage forms owing to their versatile characteristics. Recently, the preference for unique LbL assemblies in biosensor development has offered the modified performance for detection interest analyte. Beta (ß)-site amyloid precursor protein-cleaving enzyme 1 (BACE-1) is a pivotal biomarker of Alzheimer's disease (AD) and demands high sensitivity and selective identification for the early-stage diagnosis. In this work, CS-SA­platinum nanoparticles (Pt-NPs) LbL-based nanobioconjugate decorated carbon backbone-layered affinity surface plasmon resonance (Anti-BACE-1-LbL@Pt-NPs-GO-SPR) biosensor was designed for extremely sensitive and selective sensing of BACE-1. Primarily, LbL nanobioconjugate was synthesized by integrating cationic 'CS' and anionic 'SA' on the face of green-made Pt-NPs. Here, the amines of 'CS' offers a softer surface for anti-BACE-1 immobilization that leads to maintaining the bio-functionality of bioreceptors, provides the specific orientation for bioreceptors, etc. As well, the synthesized graphene oxide (GO, 2D carbon backbone) was preferred as non-plasmonic nanomaterials due to their plenty of merits in biosensors. Here, the designed biosensor provides a low detection limit (LOD) of 5.63 fg/mL and a wide linear range from 5 fg/mL to 150 ng/mL. Moreover, selectivity and real-time analyses in spiked samples exhibited their practical usefulness in complex specimens for BACE-1 detection. Hence, the decorating of antibody-immobilized CS-SA coated Pt-NPs nanobioconjugate on the face of GO has various benefits mainly extremely sensitive and superb specificity. Overall, CS and SA coated Pt-NPs bioconjugate decorated GO layered SPR biosensors can provide highly sensitive, selectivity, rapid, label-free, etc. detection of BACE-1 in clinical samples.