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Cornea-related injuries are the most common cause of blindness worldwide. Transplantation remains the primary approach for addressing corneal blindness, though the demand for donor corneas outmatches the supply by millions. Tissue adhesives employed to seal corneal wounds have shown inefficient healing and incomplete vision restoration. We have developed a biodegradable hydrogel - Kuragel, with the ability to promote corneal regeneration. Functionalized gelatin and hyaluronic acid form photo-crosslinkable hydrogel with transparency and compressive modulus similar to healthy human cornea. Kuragel composition was tuned to achieve sufficient adhesive strength for sutureless integration to host tissue, with minimal swelling post-administration. Studies in the New Zealand rabbit mechanical injury model affecting corneal epithelium and stroma demonstrate that Kuragel efficiently promotes re-epithelialization within 1 month of administration, while stroma and sub-basal nerve plexus regenerate within 3 months. We propose Kuragel as a regenerative treatment for patients suffering from corneal defects including thinning, by restoration of transparency and thickness., Competing Interests: We declare the patent application, WO2023209738A1, filed by Pandorum Technologies Pvt. Ltd., comprises information related to this work where T.B., P.A., S.K.C., and K.S. are inventors., (© 2024 Pandorum Technologies Pvt. Ltd.)

Purpose: Chemical eye injury is an acute emergency that can result in vision loss. Neurotrophic keratitis (NK) is the most common long-term manifestation of chemical injury. NK due to alkali burn affects ocular surface health and is one of its most common causes. Here, we established a rabbit model of corneal alkali burns to evaluate the severity of NK-associated changes., Material Methods: Alkali burns were induced in NZ rabbits by treating the cornea with (i) a 5 mm circular filter paper soaked in 0.75 N NaOH for 10 s (Mild NK) and (ii) trephination using a guarded trephine (5 mm diameter and 150-micron depth), followed by alkali burn, with a 5 mm circular filter paper soaked in 0.75 N NaOH for 10 s (a severe form of NK). Immediately after, the cornea was rinsed with 10 mL of normal saline to remove traces of NaOH. Clinical features were evaluated on Day 0, Day 1, Day 7, Day 15, and Day 21 post-alkali burn using a slit lamp, Pentacam, and anterior segment optical coherence tomography (AS-OCT). NK-like changes in epithelium, sub-basal nerve plexus, and stroma were observed using in vivo confocal microscopy (IVCM), and corneal sensation were measured using an aesthesiometer post alkali injury. After 21 days, pro-inflammatory cytokines were evaluated for inflammation through ELISA., Results: Trephination followed by alkali burn resulted in the loss of epithelial layers (manifested using fluorescein stain), extensive edema, and increased corneal thickness (550 µm compared to 380 µm thickness of control) evaluated through AS-OCT and increased opacity score in alkali-treated rabbit (80 compared to 16 controls). IVCM images showed complete loss of nerve fibers, which failed to regenerate over 30 days, and loss of corneal sensation-conditions associated with NK. Cytokines evaluation of IL6, VEGF, and MMP9 indicated an increased angiogenic and pro-inflammatory milieu compared to the milder form of NK and the control., Discussion: Using clinical parameters, we demonstrated that the alkali-treated rabbit model depicts features of NK. Using IVCM in the NaOH burn animal model, we demonstrated a complete loss of nerve fibers with poor self-healing capability associated with sub-basal nerve degeneration and compromised corneal sensation. This pre-clinical rabbit model has implications for future pre-clinical research in neurotrophic keratitis.

The three body fragmentation of methane dication has been studied using the technique of cold target recoil ion momentum spectroscopy. The process is initiated by impact of energetic Ar9+ ions on neutral methane and the data is subsequently collected in coincidence with Ar8+ projectile. By analysing the dissociation channels leading to (H + H+ + CH2+) and (H + H2+ + CH+) fragments, it is concluded that these fragments are formed in a sequential manner via formation of molecular intermediates CH3+ and CH2+ respectively. It is shown that these molecular intermediates carry a few eVs as their internal energies, part of which is released when they emit an H-atom with the open possibility that the final detected fragments may still be internally excited. This was accomplished by analysing the two-steps of the sequential process in their own native frames. For a molecular system having three-dimensional structure, our results prove to be an ideal example to highlight the importance of using native frames for correct interpretation of the obtained results. Our results indicate that the dissociation of methane dication can be a major source of production of H-atoms in addition to H+ fragments with the probability of the two being of similar order., (© 2023 Author(s). Published under an exclusive license by AIP Publishing.)

The three-body breakup of [C 2 H 2 ] 3+ formed in collision with Xe 9+ moving at 0.5 atomic units of velocity is studied by using recoil ion momentum spectroscopy. Three-body breakup channels leading to (H + , C + , CH + ) and (H + , H + , C 2 + ) fragments are observed in the experiment and their kinetic energy release is measured. The breakup into (H + , C + , CH + ) occurs via concerted and sequential modes, whereas the breakup into (H + , H + , C 2 + ) shows only the concerted mode. By collecting events coming exclusively from the sequential breakup leading to (H + , C + , CH + ), we have determined the kinetic energy release for the unimolecular fragmentation of the molecular intermediate, [C 2 H] 2+ . By using ab initio calculations, the potential energy surface for the lowest electronic state of [C 2 H] 2+ is generated, which shows the existence of a metastable state with two possible dissociation pathways. A discussion on the agreement between our experimental results and these ab initio calculations is presented.

We report on the direct experimental evidence of hydrogen migration in triply charged acetylene. The roaming hydrogen atom in a triply charged molecular ion is counter intuitive. The three body breakup channel C 2 H 2 3+ →H + +C + +CH + is studied using the technique of recoil ion momentum spectroscopy. The triply charged ion was generated in collisions of the neutral parent with a slow highly charged Xe 9+ ion. Three different dissociation pathways have been identified and separated, namely, concerted breakup in an acetylene configuration, concerted breakup in a vinylidene configuration, and sequential breakup via a [C 2 H] 2+ intermediate, and the branching ratio for all three pathways are determined.

The ion-induced fragmentation of CH 4 2+ into H + and CH 3 + is studied using a cold target recoil ion momentum spectroscopy in coincidence with the charge state of the post-collision projectile. Using constant velocity Ar 9+ and N 3+ , results from four different datasets are presented, with a selection on the final charge state of the projectile (Ar 8+ or Ar 7+ and N 2+ or N + ). Three distinct dissociation pathways (I, II, and III) are observed for each dataset, with the mean kinetic energy release values of around 4.7, 5.8, and 7.9 eV, respectively. The electronic states that are populated correspond to electronic configurations (1t 2 ) -2 and (2a 1 ) -1 (1t 2 ) -1 of the methane dication, CH 4 2+ . The relative branching ratios between the three pathways are discussed as a function of the charge state of the post-collision projectile, and a strong correlation with the specific nature of the ion-molecule interaction is found. The existing ab initio calculations have provided an explanation only for pathway II. In this article, we propose an explanation for pathway III, but pathway I still remains unexplained and requires further theoretical efforts. A discussion of the dependence of dissociation on the mode of excitation is presented.

Epigenetic mechanisms modulate gene expression and function without altering the base sequence of DNA. These reversible, heritable, and environment-influenced mechanisms generate various cell types during development and orchestrate the cellular responses to external stimuli by regulating the expression of genome. Also, the epigenetic modifications influence common pathological and physiological responses including inflammation, ischemia, neoplasia, aging and neurodegeneration etc. In recent past, the field of epigenetics has gained momentum and become an increasingly important area of biomedical research As far as eye is concerned, epigenetic mechanisms may play an important role in many complex diseases such as corneal dystrophy, cataract, glaucoma, diabetic retinopathy, ocular neoplasia, uveitis, and age-related macular degeneration. Focusing on the epigenetic mechanisms in ocular diseases may provide new understanding and insights into the pathogenesis of complex eye diseases and thus can aid in the development of novel treatments for these diseases. In the present review, we summarize the clinical perspective of infectious keratitis, role of epigenetics in infectious keratitis, therapeutic potential of epigenetic modifiers and the future perspective., Competing Interests: Author TB, AC, and JR are employed by the company Pandorum Technologies Ltd. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Verma, Singh, Varshney, Chandru, Acharya, Rajput, Sangwan, Tiwari, Bhowmick and Tiwari.)

Singly and multiply charged molecular ions are found in diverse environments and hold relevance for a wide range of research areas like combustion chemistry, accelerator physics, atmospheric sciences, plasma physics, astrophysics etc. Molecular dications are of special significance as they can be generated and studied comparatively easily in laboratory experiments. And they have enabled exploration of new and exciting phenomenon such as hydrogen migration, inter-atomic Coulombic decay, plasmonic excitations, orbital tomography etc. The lifetime of a molecular dication is one of its fundamental characteristics, whose measurement contributes to strengthening ab initio calculations and in predicting the concentration of its dissociation products. Most of the already reported lifetimes of molecular dications are in the range of nanoseconds to seconds and metastable states with lifetimes of the order of picoseconds have only been theoretical predicted and an experimental verification is pending. We present a method of measuring subrotational lifetimes of molecular dications formed in three-body sequential breakup of polyatomic molecular precursors. Specifically, we have measured the subrotational lifetime of [Formula: see text] , which is formed as an intermediate in the three-body sequential fragmentation of [Formula: see text]. The lifetime against dissociation is determined to be a fraction of the rotational period of [Formula: see text] and is of the order of few picoseconds. The method proposed is general and is not restricted to triatomic precursors.

A key question concerning the three-body fragmentation of polyatomic molecules is the distinction of sequential and concerted mechanisms, i.e., the stepwise or simultaneous cleavage of bonds. Using laser-driven fragmentation of OCS into O^{+}+C^{+}+S^{+} and employing coincidence momentum imaging, we demonstrate a novel method that enables the clear separation of sequential and concerted breakup. The separation is accomplished by analyzing the three-body fragmentation in the native frame associated with each step and taking advantage of the rotation of the intermediate molecular fragment, CO^{2+} or CS^{2+}, before its unimolecular dissociation. This native-frame method works for any projectile (electrons, ions, or photons), provides details on each step of the sequential breakup, and enables the retrieval of the relevant spectra for sequential and concerted breakup separately. Specifically, this allows the determination of the branching ratio of all these processes in OCS^{3+} breakup. Moreover, we find that the first step of sequential breakup is tightly aligned along the laser polarization and identify the likely electronic states of the intermediate dication that undergo unimolecular dissociation in the second step. Finally, the separated concerted breakup spectra show clearly that the central carbon atom is preferentially ejected perpendicular to the laser field.

Events from the sequential and concerted modes of the fragmentation of OCS 3+ that result in coincident detection of fragments C + , O + , and S + have been separated using a newly proposed representation. An ion beam of 1.8 MeV Xe 9+ is used to make the triply charged molecular ion, with the fragments being detected by a recoil ion momentum spectrometer. By separating events belonging exclusively to the sequential mode of breakup, the electronic states of the intermediate molecular ion (CO 2+ or CS 2+ ) involved are determined, and from the kinetic energy release spectra, it is shown that the low lying excited states of the parent OCS 3+ are responsible for this mechanism. An estimate of branching ratios of events coming from sequential versus concerted mode is presented.

The technique of recoil ion momentum spectroscopy is employed to determine the complete momentum vectors for three fragment dissociation channels, [D2O]((q+2)) → (D(+) + D(+) + O(q+)) with q = 1, 2, or 3 formed in collisions of isolated water molecules with 450 keV Xe(9 +) ions. The kinetic energy released in each of these dissociation channels is measured and angular correlations between the fragment momenta are determined. From the angular correlations of the three fragment ions with the direction of the incoming beam, a strong anisotropy in the emission of recoil fragments is reported. It is inferred that the molecular plane prefers to lie orthogonal to the incoming beam direction with certain orientations being more preferred than others and a clear signature of non-coplanar dissociation is also observed.

Members of the green fluorescent protein (GFP) family may undergo irreversible phototransformation upon irradiation with UV light. This provides clear evidence for the importance of the higher-energy photophysics of the chromophore, which remains essentially unexplored. By using time-resolved action and photoelectron spectroscopy together with high-level electronic structure theory, we directly probe and identify higher electronically excited singlet states of the isolated para- and meta-chromophore anions of GFP. These molecular resonances are found to serve as a doorway for very efficient electron detachment in the gas phase. Inside the protein, this band is found to be resonant with the quasicontinuum of a solvated electron, thus enhancing electron transfer from the GFP to the solvent. This suggests a photophysical pathway for photoconversion of the protein, where GFP resonant photooxidation in solution triggers radical redox reactions inside these proteins., (© 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

A new stability-indicating reversed-phase high-performance thin-layer chromatographic (RP-HPTLC) method for densitometric analysis of thiocolchicoside was developed and validated. The chromatograms were developed using aluminum plates pre-coated with silica gel 60 RP-18 F254S as a stationary phase and methanol : water (70 : 30 v/v) as a mobile phase. The compact band for thiocolchicoside was observed at R f value of 0.60 ± 0.02 at an absorption wavelength of 377 nm. The linear regression data for the calibration plots (r (2) = 0.9984) was found with respect to peak area in the concentration range of 100-600 ng per band. The limit of detection (LOD) and limit of quantification (LOQ) were 9.77 ng and 29.63 ng, respectively. The drug was exposed to acidic and alkaline hydrolysis, oxidation, photo degradation, and dry heat conditions. The peaks of degradation products were well-resolved from the peak of the standard drug with significantly different R f values. Statistical analysis proved that the established RP-HPTLC method is reproducible, selective, and accurate for the determination of thiocolchicoside in its formulations. The method can effectively separate the drug from its degradation products, and it can be considered as stability-indicating assay.

We report the formation of energetic neutral Rydberg hydrogen atoms and transient Rydberg molecular ions, [(H(2)O)(q+)](⋆) in ion-impact dissociation of isolated water molecules. The kinetic energy spectra of the neutral Rydberg H atoms are determined from the complete study of (H(⋆), H(+), O(+)) dissociation channel. This channel of water dissociation is suggested as a possible additional source of the energetic neutrals detected in upper atmospheres of extra solar planets, and of slow electrons which are known to play a major role in radiation induced damage to living cells., (© 2011 American Institute of Physics)

Model compounds of the green fluorescent protein (GFP) phenolate chromophore are synthesized and investigated for their intrinsic optical properties by state-of-the-art gas-phase action spectroscopy.

Photoabsorption spectra of deprotonated trans p-coumaric acid and two of its methyl substituted derivatives have been studied in gas phase both experimentally and theoretically. We have focused on the spectroscopic effect of the location of the two possible deprotonation sites on the trans p-coumaric acid which originate to either a phenoxide or a carboxylate. Surprisingly, the three chromophores were found to have the same absorption maximum at 430 nm, in spite of having different deprotonation positions. However, the absorption of the chromophore in polar solution is substantially different for the distinct deprotonation locations. We also report on the time scales and pathways of relaxation after photoexcitation for the three photoactive yellow protein chromophore derivatives. As a result of these experiments, we could detect the phenoxide isomer within the deprotonated trans p-coumaric acid in gas phase; however, the occurrence of the carboxylate is uncertain. Several computational methods were used simultaneously to provide insights and assistance in the interpretation of our experimental results. The calculated excitation energies S(0)-S(1) are in good agreement with experiment for those systems having a negative charge on a phenoxide moiety. Although our augmented multiconfigurational quasidegenerate perturbation theory calculations agree with experiment in the description of the absorption spectrum of anions with a carboxylate functional group, there are some puzzling disagreements between experiment and some calculational methods in the description of these systems.

The absorption dynamics of two model chromophores of the photoactive yellow protein were studied in gas-phase experiments. Using different time-resolving techniques with an overall sensitivity ranging from seconds down to a few nanoseconds, complex dynamics were revealed for the p -coumaric acid anion, involving both fragmentation and electron detachment as possible photoresponse channels. For the trans-thiophenyl-p-coumarate model, despite its more complex molecular structure, simpler decay dynamics showing only fragmentation were observed.

The lifetime of the SF6 anion was measured at the electrostatic ion storage ring ELISA, where decays in the time span from 100 mus to a few seconds were recorded. We find a nonexponential decay with an approximate t(-1.5) power-law dependence. The observed decay rate is accounted for by a model for thermionic emission that takes into account the initial energy spread of the SF6 molecule prior to electron capture as well as some kinetic energy of the captured electron in the applied plasma-ion source. The energy dependent decay rate is described by an Arrhenius decay constant with a pre-exponential factor and the electron affinity.

We report experimental evidence for molecular deformation due to a vibrationally active transition state of multiply charged acetylene molecules under the impact of low energy Ar8+ projectiles. "Butterflylike" structures are observed in the experimental coincidence spectra between hydrogen and carbon ionic fragments. Such structures can be generated by numerical simulations and are found to originate from the bending motion of the dissociating molecule. Angular distributions for dissociation products from triply charged C2H2 ion are reported.

We report the formation of H3+ by proton coagulation in methanol under the impact of low energy Ar(8+) projectiles. Our time-of-flight coincidence measurements with CH3OD establish that the H3+ formation arises from intramolecular bond rearrangement of the methyl group. We have performed ab initio quantum chemical calculations that show the preferred pathway for C-H3 bond cleavage. Fragmentation of organic molecules like methanol under impact of highly charged ions is suggested as an alternative mechanism of H3+ formation in outer space.