An Analytical Approach by Tri-Combination of Gradient UFLC, Response Surface Methodology and Green Chemistry Principle for Simultaneous Quantification of Azelnidipine and Chlorthalidone in Rabbit Plasma.

In this study, a sustainable and eco-friendly method is developed to quantify azelnidipine and chlorthalidone in rabbit plasma by gradient liquid chromatography based on green chemistry principle and analytical quality by design. The separation was achieved on a Shim pack C18 (25 cm × 5 cm × 4.6 µm) column with L1 packing. The mobile phase compromised of ethanol and 50-Mm ammonium acetate buffer (pH.6) at flow rate of 0.6 mL/min with 25-min runtime. The resolution and asymmetric factor were identified as critical analytical attributes (CAAs). The screening studies employing Control Noise Experimentation revealed that mobile phase pH, flow rate and ethanol concentration at 6 and 15 min significantly affected the CAAs method. The critical method parameters were optimized using Central Composition design. Chromatogram showed peak of the drugs at retention time of 9.03 min for chlorthalidone and 16.83 min for azelnidipine. The greenness score of the analytical method was found to be 1876.43 using analytical method greenness score calculator. The validation of the developed method was done which showed linearity at the range of 16-520 ng/mL, with R2 of 0.9992 and 0.9996 for azelnidipine and chlorthalidone, respectively, furthermore accuracy, precision, recovery and stability studies are carried out.

3-Polythiophene Acetic Acid Nanosphere Anchored Few-Layer Graphene Nanocomposites for Label-Free Electrochemical Immunosensing of Liver Cancer Biomarker.

This study devised a label-free electrochemical immunosensor for the quantitative detection of alpha-fetoprotein (AFP). 3-Polythiophene acetic acid (3-PTAA) nanoparticles were anchored onto a few-layer graphene (FLG) nanosheet, and the resulting nanocomposite was utilized as the immunosensor platform. The AFP antibody (anti-AFP) was immobilized on 3-PTAA@FLG via a covalent interaction between the amine group of anti-AFP and the carboxylic group of 3-PTAA via ethyl-3-(3-dimethyl aminopropyl) carbodiimide (EDC)/N-hydroxysuccinimide (NHS) coupling. FLG is largely responsible for providing electrochemical signals, whereas 3-PTAA nanoparticles are well-known for their ability to be compatible with biological molecules in neutral aqueous solutions. Moreover, the carboxyl group present in 3-PTAA effectively binds anti-AFP through EDC/NHS conjugation. Owing to good dispersibility and higher surface area of 3-PTAA, it is very convenient for casting the polymer directly on the electrode substrate followed by immobilization of anti-AFP. Thus, it is feasible to regulate the activity of AFP proteins and control the spatial distribution of the immobilized anti-AFP proteins. The electrochemical sensing performance was assessed via cyclic voltammetry and electrochemical impedance spectroscopy. For an increase in the bioconjugate concentration, the results demonstrated a surge in charge-transfer resistance and a consequent decline in the current response. This approach effectively detected AFP at an extended dynamic range of 0.0001-250 ng/mL with a detection limit of 0.047 pg/mL. Furthermore, the sensing capacity of the immunosensor for AFP detection has been demonstrated to be steady in real human serum cultures. Our approach exhibits good electrochemical performance in terms of reproducibility, selectivity, and stability, which would surely impart budding applications in the clinical diagnosis of several other tumor markers.

Asymmetric fabrication and in vivo evaluation of the wound healing potency of electrospun biomimetic nanofibrous scaffolds based on collagen crosslinked modified-chitosan and graphene oxide quantum dot nanocomposites.

Asymmetric scaffolds were developed through electrospinning by utilizing biocompatible materials for effective wound healing applications. First of all, the chitosan surface was modified with decanoyl chloride and crosslinked with collagen to synthesize collagen crosslinked modified-chitosan (CG-cross-CS-g-Dc). Then, the asymmetric scaffolds were fabricated through electrospinning, where the top layer was a monoaxial nanofiber of the PCL/graphene oxide quantum dot (GOQD) nanocomposite and the bottom layer was a coaxial nanofiber having PCL in the core and the CG-cross-CS-g-Dc/GOQD nanocomposite in the shell layer. The formation of monoaxial (∼130 ± 50 nm) and coaxial (∼320 ± 40 nm) nanofibers was confirmed by transmission electron microscopy (TEM). The presence of GOQDs contributed to antioxidant and antimicrobial efficacy. These scaffolds showed substantial antibacterial activity against the common wound pathogens Escherichia coli (E. coli) and Staphylococcus aureus (S. aureus). The scaffolds exhibited excellent cytocompatibility (MTT assay) and anti-inflammatory behaviour as analysed via the cytokine assay and biochemical analysis. The in vivo wound healing potential of the nanofibrous scaffolds was assessed with full-thickness excisional wounds in a rat model. The scaffolds accelerated the re-epithelialization as well as the collagen deposition, thereby facilitating the wound healing process in a very short span of time (10 days). Both in vitro and in vivo analyses thus provide a compelling argument for the use of these scaffolds as therapeutic biomaterials and their suitability for application in rapid wound regeneration and repair.

Biochemical Resistivity against Free Radicals and Microbes: Cooperative Action of Zn(II)/Imidazole in Phosphoesterase-Mediated Cell Death.

This work delivers a targeted synthesis of four isostructural O-substituted imidazole-based zinc(II) complexes, namely, [Zn2(L1)2(I)2](DMF) (1), [Zn2(L2)2(I)2](DMF) (2), [Zn2(L1)2(Br)2] (3), and [Zn2(L2)2(Br)2] (4), derived from homologous Schiff-base ligands HL1 and HL2 to explore their impact on free radicals, microbes, and dephosphorylation of phosphoesters. The antioxidant activity of all complexes was checked by various radical scavenging assays (ABTS+•, DPPH•, and H2O2 radical quenching). Among them, complex 2 showed superior radical quenching activity, as indicated by its lowest EC50 value and thus maximum antioxidative capability. Again, antibacterial assays against several Gram-positive and Gram-negative bacteria were conducted to evaluate the zone of inhibition. The minimum bactericidal concentration and minimum inhibitory concentration values from the microdilution method for all complexes revealed complex 3 to have maximum potency against Gram-positive bacteria. The P-O bond hydrolysis in the phospholipid chain caused by the hydrolytic phosphoesterase activity of the Zn(II)-complexes plays a crucial role in cell membrane rupture. A model substrate 4-PNPP was used to explain the potency of monomeric Zn(II) complex (3) for cell penetration over dimeric one (2) with a proper mechanism. Furthermore, a heme model substrate, Fe(TPP)Cl, has been introduced with the most potent complex 3 and has spectrophotometric evidence for covalent interaction with imidazole and Fe(III) that can disrupt the nitric oxide dioxygenase function of flavohemoglobin, leading to bacterial cell death. To our knowledge, this is the first case to report a novel mechanism of antimicrobial action where both the metal and the ligand are cooperatively involved in bacterial cell death. The main goal of this work is to invent multifunctional therapeutics as well as the proper chemical rationalization of biological processes using mechanistic approaches, which includes investigating the roles of halides, imidazoles, and solution-phase structural variations of complexes..

Hyaluronic acid-graphene oxide quantum dots nanoconjugate as dual purpose drug delivery and therapeutic agent in meta-inflammation.

Type 2 diabetes mellitus (T2DM) predominantly considered a metabolic disease is now being considered an inflammatory disease as well due to the involvement of meta-inflammation. Obesity-induced adipose tissue inflammation (ATI) is one of the earliest phenomena in the case of meta-inflammation, leading to the advent of insulin resistance (IR) and T2DM. The key events of ATI are orchestrated by macrophages, which aggravate the inflammatory state in the tissue upon activation, ultimately leading to systemic chronic low-grade inflammation and Non-Alcoholic Steatohepatitis (NASH) through the involvement of proinflammatory cytokines. The CD44 receptor on macrophages is overexpressed in ATI, NASH, and IR. Therefore, we developed a CD44 targeted Hyaluronic Acid functionalized Graphene Oxide Quantum Dots (GOQD-HA) nanocomposite for tissue-specific delivery of metformin. Metformin-loaded GOQD-HA (GOQD-HA-Met) successfully downregulated the expression of proinflammatory cytokines and restored antioxidant status at lower doses than free metformin in both palmitic acid-induced RAW264.7 cells and diet induced obese mice. Our study revealed that the GOQD-HA nanocarrier enhanced the efficacy of Metformin primarily by acting as a therapeutic agent apart from being a drug delivery platform. The therapeutic properties of GOQD-HA stem from both HA and GOQD having anti-inflammatory and antioxidant properties respectively. This study unravels the function of GOQD-HA as a targeted drug delivery option for metformin in meta-inflammation where the nanocarrier itself acts as a therapeutic agent.

Metal catalyst-free selective acetosyringone synthesis from rice straw lignin.

In recent decades, due to abundance (second most abundant natural polymer after cellulose) and sustainability, lignin has attracted much interest from different researchers to use as a raw material for producing various value-added materials such as polymer and fuel. In addition to that, the aromatic structure of lignin makes it a suitable candidate for producing platform chemicals with aromatic rings. As a result, lignin depolymerization has become an interesting process to derive different phenolic monomers like vanillin, acetosyringone, and guaiacol. Among them, due to the bioactive characteristics and efficiency of acetosyringone in plant regulatory systems, the production of acetosyringone from lignin has been presented in this work. A green and cost-effective method was developed for the selective formation of acetosyringone via depolymerization of isolated rice straw lignin (RSL) by using metal catalyst-free conditions in the biphasic medium and described. The RSL was characterized with various spectroscopic techniques such as FT-IR, solid-state 13C NMR, XPS, and TGA. The selectivity of synthesized acetosyringone during depolymerization of RSL was checked from GC-MS analysis. The molecular structure and purity of acetosyringone isolated from preparative thin layer chromatography (TLC) were confirmed with the help of 1H NMR and HRMS, respectively.

Development of an Efficient Immunosensing Platform by Exploring Single-Walled Carbon Nanohorns (SWCNHs) and Nitrogen Doped Graphene Quantum Dot (N-GQD) Nanocomposite for Early Detection of Cancer Biomarker.

In this work, a novel electrochemical immunosensor based on nitrogen doped graphene quantum dot (N-GQD) and single-walled carbon nanohorns (SWCNHs) was developed for the detection of α-fetoprotein (AFP), a cancer biomarker. Thus, to fabricate the platform of the immunosensor, nanocomposite architecture was developed by decorating N-GQD on the surface of the SWCNHs. The resulting hybrid architecture (N-GQD@SWCNHs) functioned as an exceptional base for the immobilization of antibody (Anti-AFP) through carbodiimide reaction with good stability and bioactivity. The immunosensor was prepared by evenly distributing the bioconjugates (N-GQD@SWCNHs/Anti-AFP) dispersion on the surface of the glassy carbon electrode, and subsequently blocking the remaining active sites by bovine serum albumin to prevent the nonspecific adsorption. Cyclic voltammetry and electrochemical impedance spectroscopy technique was employed to investigate the assembly process of the immunosensor. Under optimal conditions, the immunosensor exhibited a broad dynamic range in between 0.001 ng/mL to 200 ng/mL and a low detection limit of 0.25 pg/mL. Furthermore, the sensor showed high selectivity, desirable stability, and reproducibility. Measurements of AFP in human serum gave outstanding recovery within 99.2% and 102.1%. Thus, this investigation and the amplification strategy exhibited a potential role of the developed nanocomposite based sensor for early clinical screening of cancer biomarkers.

Nanotailored hyaluronic acid modified methylcellulose as an injectable scaffold with enhanced physico-rheological and biological aspects.

The collaborative endeavor in tissue engineering is to fabricate a bio-mimetic extracellular matrix to assist tissue regeneration. Thus, a novel injectable tissue scaffold was fabricated by exploring nanotailored hyaluronic acid (nHA) and methylcellulose (MC) (nHAMC) along with pristine HA based MC scaffold (HAMC). nHA with particle size ∼22 ±â€¯5.3 nm were obtained and nHAMC displayed a honeycomb-like 3D microporous architecture. Nano-HA bestowed better gel strength, physico-rheological and biological properties than HA. It creditably reduced the high content of salt to reduce the gelation temperature of MC. The properties ameliorated with increased in-corporation of nano-HA. The addition of salt showed more prominent effect on gelation temperature of nHAMC than in HAMC; and salting-out effect was dependent on nHA/HA content. Biocompatible nHAMC assisted adequate cell adherence and proliferation with more extended protrusions with better migration rate than control. Thus, biomodulatory effect of nanotailored glycosaminoglycan could be asserted to design an efficient thermo-responsive scaffold.

Sustained release of ketorolac tromethamine from poloxamer 407/cellulose nanofibrils graft nanocollagen based ophthalmic formulations.

There has been extensive utilization of poloxamer 407 (PM) for the delivery of various ophthalmic drugs aimed at efficient ophthalmic drug delivery approach for longer precorneal residence time along with acceptable bioavailability of drugs. We have studied the effect of nanocellulose grafted collagen (CGC) on the performance of in situ gels based on PM for the controlled in vitro release of Ketorolac Tromethamine (KT). CGC has shown great influence evident by the reduction in PM critical gelation concentration, increased gel strength, and prolonged the release of loaded drugs compared with the virgin PM gel. The engineered nanocomposite formulations established an anomalous diffusion mechanism along with a Fickian diffusion controlled drug release for 1.5 & 1.75 w/v% CGC reinforced PM. Hence, the synthesized in situ nanocomposites are potential candidates for ophthalmic drug delivery system.

Optical Coherence Tomography Findings in Patients of Parkinson's Disease: An Indian Perspective.

BACKGROUND: Parkinson's disease (PD) is one of the most common neurodegenerative movement disorders and its incidence is increasing worldwide along with population aging. Previous clinical and histologic studies suggest that the neurodegenerative process, which affects the brain, may also affect the retina of PD patients. OBJECTIVE: The objective of this study was to determine the thickness changes of retina nerve fibers and macular volume with optical coherence tomography (OCT) in PD patients. MATERIALS AND METHODS: The spectral domain OCT was used to assess the thickness of retinal nerve fiber layer (RNFL) and macular volume from 34 PD patients and 50 healthy age-matched controls. RESULTS: Compared with healthy age-matched controls, the RNFL thickness of PD patients was much thinner (P < 0.05) in all retinal quadrants, with temporal thinning being more than nasal thinning. Macular volumes were diminished in both perifoveal and outer macular regions in all sectors (P < 0.05) with preserved foveal volume. The degree of tissue loss corroborated with the severity of disease as objectively assessed by standardized rating scales (UPDRS). CONCLUSION: There is generalized retinal nerve degeneration in patients of PD and the degree of loss correlated with the severity and duration of disease.

Tailoring the Efficacy of Multifunctional Biopolymeric Graphene Oxide Quantum Dot-Based Nanomaterial as Nanocargo in Cancer Therapeutic Application.

Nanotechnology has acquired an immense recognition in cancer theranostics. Considerable progress has been made in the development of targeted drug delivery system for potent delivery of anticancer drugs to tumor-specific sites. Recently, multifunctional nanomaterials have been explored and used as nanovehicles to carry drug molecules with enhanced therapeutic efficacy. In this present work, graphene oxide quantum dot (GOQD) was conjugated with folic acid functionalized chitosan (FA-CH) to develop a nanocargo (FA-CH-GOQD) for drug delivery in cancer therapy. The synthesized nanomaterials were characterized using Fourier transform infrared spectroscopy, ultraviolet-visible spectroscopy, scanning electron microscopy, transmission electron microscopy, and dynamic light scattering. Photoluminescence spectroscopy was also employed to characterize the formation of GOQD. To validate the efficacy of FA-CH-GOQD as nanocarriers, doxorubicin (DOX) drug was chosen for encapsulation. The in vitro release pattern of DOX was examined in various pH ranges. The drug release rate in a tumor cell microenvironment at pH 5.5 was found higher than that under a physiological range of pH 6.5 and 7.4. An MTT assay was performed to understand the cytotoxic behavior of GOQD and FA-CH-GOQD/DOX. Cytomorphological micrographs of the A549 cell exhibited the various morphological arrangements subject to apoptosis of the cell. Cellular uptake studies manifested that FA-CH-GOQD could specifically transport DOX within a cancerous cell. Further anticancer efficacy of this nanomaterial was corroborated in a breast cancer cell line and demonstrated through 4',6-diamidino-2-phenylindole dihydrochloride staining micrographs.

Analysis of physico-chemical characteristics and metals in water sources of chromite mining in Sukinda Valley, Odisha, India.

The extraction of chromite is being carried out since 1950 in Sukinda valley region of Odisha in India. Different physico-chemical parameters and level of metals in groundwater (tube well and dug well), surface water (Damsal nala), mining drainage water and bottom sediment of Damsal nala were analysed. The results revealed that the total Cr content of groundwater (0.04-0.07 ppm), Damsal nala (0.10 - 0.20 ppm) and mine drainage water (20.12-56.51 ppm) exceeded the permissible limit in all seasons. The Cr (VI) content of mine drainage water exceeded the permissible limit (0.05 ppm as per the recommendation of WHO). The pH (6.5-7.9), available N (6.27-18.82 kg ha(-1)), available PO4(-3) (15.39-123.11 kg ha(-1)), available K+ (21.07-410.89 kg ha(-1)) and organic matter (0.20-1.55%) content of bottom sediment of Damsal nala varied seasonally and the lowest values were found during monsoon. The physico-chemical characteristics of water from Damsal nala, groundwater and mine drainage, like COD, total Fe, TDS, TSS, F, available PO4(-3), available K+ etc., including pH are a serious concerned in this study area.

Tackling hepatitis B virus-associated hepatocellular carcinoma--the future is now.

Hepatocellular carcinoma (HCC) is one of the most lethal and prevalent cancers in many developing countries including India. Among the various etiological factors being implicated in the cause of HCC, the most important cause, however, is hepatitis B virus (HBV) infection. Among all HBV genes, HBx is the most critical carcinogenic component, the molecular mechanisms of which have not been completely elucidated. Despite its clinical significance, there exists a very elemental understanding of the molecular, cellular, and environmental mechanisms that drive disease pathogenesis in HCC infected with HBV. Furthermore, there are only limited therapeutic options, the clinical benefits of which are insignificant. Therefore, the quest for novel and effective therapeutic regimen against HBV-related HCC is of paramount importance. This review attempts to epitomize the current state of knowledge of this most common and dreaded liver neoplasm, highlighting the putative treatment avenues and therapeutic research strategies that need to be implemented with immediate effect for tackling HBV-related HCC that has plagued the medical and scientific fraternity for decades. Additionally, this review proposes a novel "five-point" management algorithm for HBV-related HCC apart from portraying the unmet needs, principal challenges, and scientific perspectives that are relevant to controlling this accelerating global health crisis.

The α(1)AT and TIMP-1 Gene Polymorphism in the Development of Asthma.

Asthma has been an inflammatory disorder accompanied by tissue remodeling and protease-antiprotease imbalance in lungs. The SNPs of alpha-1 antitrypsin (α(1)AT) and tissue inhibitor of metalloproteinase-1 (TIMP-1) genes were studied for their association with asthma. Genotyping of α(1)AT and TIMP-1 genes was performed in 202 asthmatics and 204 controls. Serum levels of α(1)AT, TIMP-1 and cytokines were estimated to find if the interplay between genotypes and cellular biomarkers determines the pathogenesis of asthma. The analysis of results showed significantly low level of α(1)AT in the serum of asthmatics as compared to controls (P = 0.001), whereas cytokines were elevated in patients. No significant difference was observed in the concentration of TIMP-1 in patients and controls. Genotyping led to the identification of 3 SNPs (Val213Ala, Glu363Lys, and Glu376Asp) in α(1)AT gene. The novel SNP Glu363Lys of α(1)AT was found to be associated with asthma (P = 0.001). The analysis of TIMP-1 gene showed the occurrence of seven SNPs, including a novel intronic SNP at base G3774A. The allele frequency of G3774A and Phe124Phe was significantly higher in asthmatics as compared to controls. Thus, the SNP Glu363Lys of α(1)AT and G3774A and Phe124Phe of TIMP-1 could be important genetic markers for use in better management of the disease.

Combinatorial effect of TIMP-1 and α1AT gene polymorphisms on development of chronic obstructive pulmonary disease.

OBJECTIVE: To study the role of α(1)AT and TIMP-1 gene polymorphisms in development of COPD. DESIGN AND METHODS: Blood samples from total 408 subjects (217 COPD patients and 191 controls) were used for genotyping and estimating biolevels of α(1)AT, TIMP-1 and inflammatory cytokines. Data was analyzed to determine the role of interaction of TIMP-1 and α(1)AT genes; and interplay between various genotypes and biolevels of α(1)AT, TIMP-1 and inflammatory cytokines in development of COPD. RESULTS: Significantly low levels of α(1)AT and TIMP-1 were observed in COPD patients as compared to controls (P = 0.001), where as the inflammatory cytokines were found to be increased in patients. PIM3 allele of α(1)AT gene in COPD patients was found to be associated with low levels of α(1)AT (P = 0.001), the effect being more pronounced when PIM3 combined with rs6609533 of TIMP-1 gene (P = 0.0001). Combination of genotypes rs6609533 of TIMP-1 and PIM3 of α(1)AT containing the risk alleles was over-represented in patients (P = 0.005). CONCLUSION: The SNP rs6609533 of TIMP-1 gene interacted with PIM3 of α(1)AT to make a possible risk combination for development of COPD.

Rapid vertical mixing rates in deep waters of the Andaman Basin.

The Andaman Basin is an enclosed region in the northeastern Indian Ocean with its deep water below approximately 1800 m almost isolated with respect to horizontal ventilation by the Andaman-Nicobar Islands separating it from the Bay of Bengal. The physical and chemical properties including radiocarbon ((14)C) measured at two stations of the Andaman Basin show negligible variation with depth in the waters below 1300 m, indicating a well-mixed water mass. This study attempts to derive the mixing rates for deep waters of the Andaman Basin. Model calculations based on (14)C profile measurements indicate rapid vertical mixing (vertical advection velocity, w>200 m year(-1)) in waters deeper than 1800 m of the basin. For a basin with deep water thickness of 1000 m below 1800 m, deduced mixing rate of >200 m year(-1) translates to mixing time of <5 years. As shown for other regions, the possible mechanism responsible for such high vertical mixing rates could be the internal waves generated from tidal currents flowing through rough topography. In addition, Andaman Basin is underlain with a young crust and is known for its high heat flow, which also could contribute to the high vertical mixing.