Synthesis of Triazolo[4',5':4,5]furo[2,3-c]pyridine via Post Modification of an Unusual Groebke-Blackburn-Bienaymé Multicomponent Reaction.

The Groebke-Blackburn-Bienaymé (GBB) reaction is a well-established three-component reaction for synthesizing imidazofused scaffolds from heterocyclic amidines, aldehydes, and isonitriles. However, the replacement of pyridoxal as an aldehyde component in this reaction results in the formation of the furo[2,3-c]pyridine skeleton as an "unusual GBB product". Despite the interesting nature of this unusual reaction, not much work was further reported. The present research investigates the optimization strategy for the synthesis of novel tricyclic triazolo[4',5':4,5]furo[2,3-c]pyridines via diazotization of 2,3-diamino-furo[2,3-c]pyridines specifically synthesized utilizing the chemistry of tert-alkyl isocyanide.

The Electroencephalographic Brainwave Spectrum, Mindful Meditation, and Awareness: Hypothesis.

It is hypothesized that being mindfully aware is a spontaneous state of being. It is imbued with joy, peace and happiness. Such a state is periodically revealed during restful attentiveness or presence. It is also associated with spontaneous brain alpha activity of 8-13 Hz. In deep nonrapid eye movement sleep, there is synchronous delta activity at a coherent frequency of 0.1 Hz. Both of these brainwave ground states are spontaneous, calm and effortless. When any physical or mental effort is made, the alpha rhythm is desynchronized, and it is superposed by faster brain waves of beta (13-30 Hz) and gamma frequencies (30-150 Hz). This is associated with a stream of dualistic conscious experiences with contents. During deep sleep, delta activity is superposed by beta and gamma activity with microarousals resulting in dream experiences. During effortless, meditative awareness, the whole family of alpha rhythm is synchronized including (a) Occipital-parietal alpha with visual clarity, formless color, and the absence of visual imagery (b) Frontal eye-field alpha with relatively motionless eyes, and the absence of voluntary actions or plans to move the eyes in some direction, along with nonactive working memory, (c) Somatosensory alpha or Mu rhythm from the somatic motor-sensory cortex with the resultant stillness of the body including head, face, larynx, spine, hands and legs, (d) Mid-temporal auditory alpha with vocal quietness and internal verbal silence (Maunam) with a feeling of spontaneous silence and serenity, (e) Cingulate and precuneus alpha resulting in freedom from autobiographical memories and the sense of agency or ego. The insular cortex serves as a gatekeeper, a hierarchical controller to switch between conscious engagement or disengagement from the internal or the external world. It switches between the default mode network and the executive frontoparietal networks, between the sequential and the parallel modes of functioning. Mindful consciousness is local and dualistic, whereas mindful awareness is nonlocal and nondual.

Discovery of LNP1892: A Precision Calcimimetic for the Treatment of Secondary Hyperparathyroidism.

The calcium sensing receptor (CaSR) plays an important role in maintaining calcium homeostasis. The use of calcimimetic cinacalcet has been established to activate CaSR and normalize hypercalcemia. However, cinacalcet has limitations due to its high cLogP and pKa. A systematic optimization of cinacalcet to reduce its cLogP and pKa yielded compound 23a (LNP1892). Compound 23a showed excellent potency and a favorable pharmacokinetics profile, and lacked the liabilities of cinacalcet, making it a highly differentiated precision calcimimetic. In adenine-diet-induced chronic kidney disease (CKD) models, 23a demonstrated robust and dose-dependent efficacy, as measured by plasma parathyroid hormone (PTH) levels. It also showed an excellent safety profile in animal studies. Phase 1 clinical trials with 23a in healthy volunteers confirmed its excellent safety, tolerability, and effectiveness in lowering PTH levels in a dose-dependent manner, without causing symptomatic hypocalcaemia. Encouraged by these promising results, LNP1892 was taken to a Phase 2 study in CKD patients.

Design, synthesis, molecular dynamics simulation, MM/GBSA studies and kinesin spindle protein inhibitory evaluation of some 4-aminoquinoline hybrids.

The discovery of novel chemotherapeutic agents is always challenging for researchers in industry and academia. Among the recent promising anticancer therapeutic targets, an important modulatory factor in mitosis is the expression of the kinesin family motor protein (Eg5). In terms of chemotherapy treatment, mitosis has gained significant attention due to its role as one of the biological processes that can be intervened in it. This study was undertaken to design, synthesise and evaluation of 4-aminoquinoline hybrid compounds as potential Eg5 inhibitors. Based on data collected from Malachite green and steady state ATPase assays, it has been determined that compounds such as 6c, 6d, 6g, and 6h are sensitive to Eg5 inhibition. In special mention, compounds 4 and 6c showed promising inhibitory activity in Malachite green assay with IC50 values of 2.32 ± 0.23 µM and 1.97 ± 0.23 µM respectively. Compound 4 showed favourable inhibitory potential Steady state ATPase Assay with IC50 value of 5.39 ± 1.39 µM. We performed molecular docking, MM/GBSA calculations, and molecular dynamic simulations to evaluate the interactions between ligands and the binding site of the kinesin spindle protein to evaluate the functional consequences of these interactions. As a result of these findings, it can be concluded that these 4-amioquinoline Schiff's base hybrids may prove to be promising candidates for development as novel inhibitors of Eg5. Further in-vivo research in this area is required.

Diversity in the ligand binding pocket of HapR attributes to its uniqueness towards several inhibitors with respect to other homologues - A structural and molecular perspective.

Vibrio cholerae is a prolific bacterium. Cumulative studies clearly demonstrate the key role of quorum sensing on the lifecycle of this bacterium. Of the sensory network components, HapR is known as high cell density master regulator. Until now, no information is available on native HapR ligand despite the protein having a ligand binding pocket. Interestingly, function of SmcR, a HapR homologue of Vibrio vulnificus is inhibited by a small molecule Qstatin. Structural analysis of SmcR with Qstatin identifies key interacting residues in SmcR ligand binding domain. Despite bearing significant homology with SmcR, HapR function remained unabated by Qstatin. Sequence alignment indicates divergence in the key residues of ligand binding pocket between these two regulators. A series of ligand binding domain mutants of HapR was constructed where only HapR quadruple mutant responded to Qstatin and newly synthesized IMT-VC-212. Crystal structure analysis revealed four key residues are responsible for changes in the volume of ligand binding pocket of HapR quadruple mutant compared to the wild type counterpart, thereby increasing the accessibility of Qstatin and its derivative in case of the former. The mechanistic insights exuberating from this study will remain instrumental in designing inhibitors against wild type HapR.

Biophysical study of cisplatin loaded albumin-gold nanoparticle and its interaction with glycans of gp60 receptor.

The biophysical study provides a quantitative understanding of biomolecular interaction. The interaction of protein-nanoparticle has been critically examined using various biophysical and biochemical tools. The present investigation focussed on the biophysical characterization of anticancer drug cisplatin (CPT) with Bovine Serum Albumin (BSA) - Gold nanoparticles (GNP) conjugate; and BSA-CPT-GNP interaction with glycan sugars of glycoprotein receptor. Spectroscopic study (UV visible and fluorescence) showed strong binding of CPT loaded BSA with GNP. The binding between BSA-CPT-GNP and glycan sugars of gp60 receptor was estimated. Circular Dichroism (CD) spectroscopy study revealed weak alteration in the secondary structure of BSA upon CPT and GNP binding. Dynamic Light Scattering (DLS) data indicated the changes in the size of conjugates; zeta potential data showed the stability of conjugates. Biocompatible studies showed no toxicity to RBCs and chorioallantoic membrane (CAM). The mechanisms of interaction have been explored at the molecular and cellular levels. This investigation can be effectively extrapolated for in-vivo and in-vitro targeted drug delivery studies for cancer therapy.

One-Pot Domino Reaction: Direct Access to Polysubstituted 1,4-Benzothiazine 1,1-Dioxide via Water-Gas Shift Reaction Utilizing DMF/H2O.

Benzothiazine 1,1-dioxide (BTDO) is a privileged chemical motif, and its metal-free domino access is in high demand. Current BTDO production methods require costly metal catalysts or harsh reaction conditions. A facile domino approach to BTDO via a water-gas shift reaction (WGSR) employing sodium 2-nitrobenzenesulfinates and α-bromo ketones is presented. This strategy is cost-effective and environmentally beneficial. The optimized reaction conditions demonstrated remarkable chemical tolerance to a wide range of electrically and sterically varied substituents on both coupling partners.

Consciousness, Awareness, and Presence: A Neurobiological Perspective.

It is proposed that consciousness is different from awareness. Consciousness can be thought of as a dualistic, embodied, and embedded cognitive process, whereas awareness is a nondual and nonlocal process. Nonlocal awareness is the ever-present, ever-fresh, and an affective self-awareness that can be aware of itself as well as of the ongoing subject-object duality, and cognitive conscious contents. This nonlocal awareness is our default mode state. Although very few of us are aware of it due to our habitual mental preoccupation and mind-wandering. We need to relax, learn to meditate, let go of all preoccupations, and return to our default mode state of being, which is peaceful, silent, fulfilling, energetic, and ever-fresh. Then, one feels effortlessly alive and free and at home in the world. This is the essence of meditation for living a happy, peaceful, and meaningful life. The rest of the article provides details of meditative presence, yoga meditation, and mindfulness meditation with their current practice and applications. The main focus of the article is on the neurobiology of meditation, which is discussed in detail. It covers the experientially perceived mind-space including personal, peripersonal, and extrapersonal space, the concepts of mind in the Western and Eastern literature, and the neurobiological foundation in the brain stem, reticular-limbic system, forebrain including the five thalamo-cortical-basal ganglia circuits, multiple sensory modalities, integrated perception, speech production, language communication, voluntary movements, and intentional actions. The wholeness of conscious mind is expressed as bio-psycho-social-abstract/spiritual.

Role of Oxidative Stress and the Protective Effect of Fermented Papaya Preparation in Sickle Cell Disease.

Fermented papaya preparation (FPP) is the source of antioxidants that may help in reducing the complications associated with oxidative stress and may improve the quality of life in sickle cell disease patients. In this study, we assessed the in vitro effect of FPP on sickled red blood cells (RBCs) using oxidative stress markers and observed that FPP has the potential to reduce the oxidative stress. Scanning electron microscopy (SEM) and eosin 5' malaemide (E5'M) dye test showed that FPP protects red cell morphology against the oxidative stress. Liquid chromatography mass spectrometry (LCMS) analysis of FPP suggests the presence of essential amino acids, vitamin D3, and its derivatives. Fermented papaya preparation can be of benefit either in reducing oxidative stress parameters or in preventing pathophysiological events in the sickle cell disease patients.

Cationic Chitosan/Pectin Polyelectrolyte Nanocapsules of Moxifloxacin as Novel Topical Management System for Bacterial Keratitis.

PURPOSE: Moxifloxacin (MOX) is a fourth-generation fluoroquinolone and a broad spectrum antibiotic used in the management of bacterial keratitis (BK). This investigation aimed to formulate MOX-loaded chitosan/pectin cationic polyelectrolyte nanocapsules (CPNCs) for the effective topical treatment of BK. METHODS: Physicochemical properties like nanocapsule size, charge, drug entrapment efficiency (EE), viscosity, pH, and in-vitro release profile of CPNCs were evaluated. The in-vitro antibacterial activity of CPNCs and marketed formulations (MFs) was studied against Staphylococcus aureus. Ex-vivo corneal permeation studies of CPNCs were evaluated with the help of a modified diffusion apparatus, which was used with goat cornea. The pharmacodynamic study was performed with optimized CPNCs on a BK-induced rabbit eye model and compared with MF. RESULTS: The optimized nanocapsules appeared as positive charge (+19.91 ± 0.66) with a nano size (242.0 ± 0.30 nm) as calculated by the dynamic light scattering method. The in-vitro release profile of CPNCs exhibited sustained release properties. The ex-vivo permeation pattern also supported the improved drug permeation through the cornea from CPNCs as compared with MF. Draize irritation studies confirmed that the prepared formulation is compatible with the corneal tissue. The in-vivo study concluded that the antibacterial activity of CPNCs was improved when evaluated with MF. CONCLUSION: The obtained results showed that CPNCs were the better choice for the management of BK therapy due to its capability to improve the corneal adhesion of CPNCs through direct interaction with the mucous membrane of the corneal tissue.

Severity Prediction over Parkinson's Disease Prediction by Using the Deep Brooke Inception Net Classifier.

Parkinson's disease (PD) is a neurodegenerative illness that progresses and is long-lasting. It becomes more difficult to talk, write, walk, and do other basic functions when the brain's dopamine-generating neurons are injured or killed. There is a gradual rise in the intensity of these symptoms over time. Using Parkinson's Telemonitoring Voice Data Set from UCI and deep neural networks, we provide a strategy for predicting the severity of Parkinson's disease in this research. An unprocessed speech recording contains a slew of unintelligible data that makes correct diagnosis difficult. Therefore, the raw signal data must be preprocessed using the signal error drop standardization while the features can be grouped by using the wavelet cleft fuzzy algorithm. Then the abnormal features can be selected by using the firming bacteria foraging algorithm for feature size decomposition process. Then classification was made using the deep brooke inception net classifier. The performances of the classifier are compared where the simulation results show that the proposed strategy accuracy in detecting severity of the Parkinson's disease is better than other conventional methods. The proposed DBIN model achieved better accuracy compared to other existing techniques. It is also found that the classification based on extracted voice abnormality data achieves better accuracy (99.8%) over PD prediction; hence it can be concluded as a better metric for severity prediction.

Free spermidine evokes superoxide radicals that manifest toxicity.

Spermidine and other polyamines alleviate oxidative stress, yet excess spermidine seems toxic to Escherichia coli unless it is neutralized by SpeG, an enzyme for the spermidine N-acetyl transferase function. Thus, wild-type E. coli can tolerate applied exogenous spermidine stress, but ΔspeG strain of E. coli fails to do that. Here, using different reactive oxygen species (ROS) probes and performing electron paramagnetic resonance spectroscopy, we provide evidence that although spermidine mitigates oxidative stress by lowering overall ROS levels, excess of it simultaneously triggers the production of superoxide radicals, thereby causing toxicity in the ΔspeG strain. Furthermore, performing microarray experiment and other biochemical assays, we show that the spermidine-induced superoxide anions affected redox balance and iron homeostasis. Finally, we demonstrate that while RNA-bound spermidine inhibits iron oxidation, free spermidine interacts and oxidizes the iron to evoke superoxide radicals directly. Therefore, we propose that the spermidine-induced superoxide generation is one of the major causes of spermidine toxicity in E. coli.

Role of toll-like receptor 7/8 pathways in regulation of interferon response and inflammatory mediators during SARS-CoV2 infection and potential therapeutic options.

Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV2) is the causative agent of Corona Virus Disease 2019 (COVID-19). Lower production of type I and III interferons and higher levels of inflammatory mediators upon SARS-CoV2 infection contribute to COVID-19 pathogenesis. Optimal interferon production and controlled inflammation are essential to limit COVID-19 pathogenesis. However, the aggravated inflammatory response observed in COVID-19 patients causes severe damage to the host and frequently advances to acute respiratory distress syndrome (ARDS). Toll-like receptor 7 and 8 (TLR7/8) signaling pathways play a central role in regulating induction of interferons (IFNs) and inflammatory mediators in dendritic cells. Controlled inflammation is possible through regulation of TLR mediated response without influencing interferon production to reduce COVID-19 pathogenesis. This review focuses on inflammatory mediators that contribute to pathogenic effects and the role of TLR pathways in the induction of interferon and inflammatory mediators and their contribution to COVID-19 pathogenesis. We conclude that potential TLR7/8 agonists inducing antiviral interferon response and controlling inflammation are important therapeutic options to effectively eliminate SARS-CoV2 induced pathogenesis. Ongoing and future studies may provide additional evidence on their safety and efficacy to treat COVID-19 pathogenesis.

Peroxynitrite-induced conformational changes in DNA that lead to cell death: UV, CD spectral, molecular dynamics simulation and FACS analysis.

Peroxynitrite is known to react with biomolecules leading to their structural and function alteration. Structural alteration in DNA induced by peroxynitrite is not clearly known. The current study was carried out to decipher the changes induced by peroxynitrite using UV-Vis spectra, circular dichrometry, molecular dynamics simulation followed by restriction digestion. Apoptotic markers Bax, Bcl-2 and caspase genes were also studied by FACS in peroxynitrite induced PC12 cells. The results obtained showed that PXN binds to DNA leading to hyperchromicity of DNA in the presence of PXN over a period of time and the same was established by In silico studies where PXN modifies the DNA to accommodate itself into the stacking and brings about the significant structural alterations. Further, FACS studies reveal that Bcl-2 gene expression was down regulated whereas BAXand caspase genes were up regulated compared to control concluding that PXN induces apoptotic cell death in PC12 cells.

Terminal Respiratory Oxidases: A Targetables Vulnerability of Mycobacterial Bioenergetics?

Recently, ATP synthase inhibitor Bedaquiline was approved for the treatment of multi-drug resistant tuberculosis emphasizing the importance of oxidative phosphorylation for the survival of mycobacteria. ATP synthesis is primarily dependent on the generation of proton motive force through the electron transport chain in mycobacteria. The mycobacterial electron transport chain utilizes two terminal oxidases for the reduction of oxygen, namely the bc1-aa3 supercomplex and the cytochrome bd oxidase. The bc1-aa3 supercomplex is an energy-efficient terminal oxidase that pumps out four vectoral protons, besides consuming four scalar protons during the transfer of electrons from menaquinone to molecular oxygen. In the past few years, several inhibitors of bc1-aa3 supercomplex have been developed, out of which, Q203 belonging to the class of imidazopyridine, has moved to clinical trials. Recently, the crystal structure of the mycobacterial cytochrome bc1-aa3 supercomplex was solved, providing details of the route of transfer of electrons from menaquinone to molecular oxygen. Besides providing insights into the molecular functioning, crystal structure is aiding in the targeted drug development. On the other hand, the second respiratory terminal oxidase of the mycobacterial respiratory chain, cytochrome bd oxidase, does not pump out the vectoral protons and is energetically less efficient. However, it can detoxify the reactive oxygen species and facilitate mycobacterial survival during a multitude of stresses. Quinolone derivatives (CK-2-63) and quinone derivative (Aurachin D) inhibit cytochrome bd oxidase. Notably, ablation of both the two terminal oxidases simultaneously through genetic methods or pharmacological inhibition leads to the rapid death of the mycobacterial cells. Thus, terminal oxidases have emerged as important drug targets. In this review, we have described the current understanding of the functioning of these two oxidases, their physiological relevance to mycobacteria, and their inhibitors. Besides these, we also describe the alternative terminal complexes that are used by mycobacteria to maintain energized membrane during hypoxia and anaerobic conditions.

Discovery of a Potent and Selective PI3Kδ Inhibitor (S)-2,4-Diamino-6-((1-(7-fluoro-1-(4-fluorophenyl)-4-oxo-3-phenyl-4H-quinolizin-2-yl)ethyl)amino)pyrimidine-5-carbonitrile with Improved Pharmacokinetic Profile and Superior Efficacy in Hematological Cancer Models.

PI3Kδ inhibitors have been approved for B-cell malignancies like CLL, small lymphocytic lymphoma, and so forth. However, currently available PI3Kδ inhibitors are nonoptimal, showing weakness against at least one of the several important properties: potency, isoform selectivity, and/or pharmacokinetic profile. To come up with a PI3Kδ inhibitor that overcomes all these deficiencies, a pharmacophoric expansion strategy was employed. Herein, we describe a systematic transformation of a "three-blade propeller" shaped lead, 2,3-disubstituted quinolizinone 11, through a 1,2-disubstituted quinolizinone 20 to a novel "four-blade propeller" shaped 1,2,3-trisubstituted quinolizinone 34. Compound 34 has excellent potency, isoform selectivity, metabolic stability across species, and exhibited a favorable pharmacokinetic profile. Compound 34 also demonstrated a differentiated efficacy profile in human germinal center B and activated B cell-DLBCL cell lines and xenograft models. Compound 34 qualifies for further evaluation as a candidate for monotherapy or in combination with other targeted agents in DLBCLs and other forms of iNHL.

Role of MMP-2 and its inhibitor TIMP-2 as biomarkers for susceptibility to systemic lupus erythematosus.

Aim: To investigate the possible association between MMP-2 (-1575 G/A, -1306 C/T) and its inhibitor TIMP-2 (-418 G/C) functional polymorphisms with development of severity in systemic lupus erythematosus (SLE) patients. Materials & methods: 150 SLE patients and matched healthy controls were recruited. Polymorphisms were detected by PCR-RFLP and serum levels by ELISA. Results: Mean MMP-2 and TIMP-2 serum level and mRNA expression were significantly increased in SLE cases as compared with controls (p < 0.0001). The concomitant presence of both MMP-2 1575A and its inhibitor TIMP-2 418C alleles synergistically increased the risk of SLE by 3.25-fold (CI: 1.44-7.34, p = 0.003). Conclusion: MMP-2, TIMP-2 and MMP-2/TIMP-2 ratios may act as biomarkers for susceptibility to SLE.

Biophysical interactions between silver nanoparticle-albumin interface and curcumin.

Active targeted drug delivery methods facilitate effective uptake of functionalized nanoparticles through receptor-mediated transcytosis. In recent years, albumin-nanoparticle interaction has been critically examined so that this functionalized nanoparticle can be efficiently loaded with drugs. The present investigation aims at understanding the adsorption of Bovine Serum Albumin (BSA) on Silver Nanoparticle (SNP) surface, preparation of soft conjugates (SC) and hard conjugates (HC) of BSA-functionalized SNP (SNP-BSA), and their interaction with curcumin (CUR). HC contains tightly bound BSA whereas SC involves tightly and loosely bound BSA. Increase in the hydrodynamic radii of conjugates was observed upon SNP incubation with increased concentration of BSA. Three different SNP-BSA conjugate ratios were selected to study their interaction with CUR. Fluorescence spectroscopy showed a strong association between CUR and SNP:BSA conjugates. However, binding varied with a change in the conjugate ratio. Circular Dichroism (CD)/Fourier Transform Infrared (FTIR) spectroscopy revealed the alterations in the secondary structure of BSA upon CUR binding to the conjugates. Zeta potential data indicated stable conjugate formation. CUR in SNP:BSA conjugate was found to have a higher half-life as compared to the control. We believe that this is the first biophysical characterization report of conjugates that can be effectively extrapolated for targeted drug delivery.

Phosphoproteomic analysis identifies CLK1 as a novel therapeutic target in gastric cancer.

BACKGROUND: Phosphorylation is an important regulatory mechanism of protein activity in cells. Studies in various cancers have reported perturbations in kinases resulting in aberrant phosphorylation of oncoproteins and tumor suppressor proteins. METHODS: In this study, we carried out quantitative phosphoproteomic analysis of gastric cancer tissues and corresponding xenograft samples. Using these data, we employed bioinformatics analysis to identify aberrant signaling pathways. We further performed molecular inhibition and silencing of the upstream regulatory kinase in gastric cancer cell lines and validated its effect on cellular phenotype. Through an ex vivo technology utilizing patient tumor and blood sample, we sought to understand the therapeutic potential of the kinase by recreating the tumor microenvironment. RESULTS: Using mass spectrometry-based high-throughput analysis, we identified 1,344 phosphosites and 848 phosphoproteins, including differential phosphorylation of 177 proteins (fold change cut-off ≥ 1.5). Our data showed that a subset of differentially phosphorylated proteins belonged to splicing machinery. Pathway analysis highlighted Cdc2-like kinase (CLK1) as upstream kinase. Inhibition of CLK1 using TG003 and CLK1 siRNA resulted in a decreased cell viability, proliferation, invasion and migration as well as modulation in the phosphorylation of SRSF2. Ex vivo experiments which utilizes patient's own tumor and blood to recreate the tumor microenvironment validated the use of CLK1 as a potential target for gastric cancer treatment. CONCLUSIONS: Our data indicates that CLK1 plays a crucial role in the regulation of splicing process in gastric cancer and that CLK1 can act as a novel therapeutic target in gastric cancer.

Effect of Smartphone Light Fluxes on Cornea: A Biophysical Study.

OBJECTIVE: Biophysical study to investigate (a) the effects of smartphone light fluxes (SPLF) on isolated mammalian cornea and model protein (insulin), (b) to predict the possible visual interference of SPLF. MATERIALS AND METHODS: Fresh goat cornea and insulin protein were used as an experimental model system. The energy of absorbed SPLF was measured using chemical dosimeter. The effect of SPLF on the aggregation of model protein was studied using fluorescence spectroscopy and dynamic light scattering (DLS). Fluorescence microscopy, scanning electron microscopy (SEM), DLS, were used for cornea imaging. RESULTS: The spectral emission peak of SPLF was observed at 380 nm and 420 nm. Absorbed radiation of SPLF was found to be 2.82 mWm-2 and 1.92 mWm-2 for collimated (focussed) and noncollimated (nonfocussed) condition, respectively. Secondary structural changes of insulin were observed by fluorescence and zeta potential after SPLF exposure. SEM study revealed the disorganization of the epithelial cell surface, increase in intercellular space, disorganization of primary epithelium layer, and exposure of the second layer is seen in depth. Differential Interference Microscopy showed an optical gradient in images that appears to be changed in specimen structure. Fluorescence microscopy showed disorganization in epithelial cell pattern. A significant difference in bio-molecular permeation was observed in the exposed cornea. Ultraviolet UV-visible spectroscopy study indicated a reduction in light transmission through the cornea. CONCLUSIONS: The obtained results indicate changes in physicochemical and morphological modifications in the cornea and insulin modifications after exposed to SPLF.