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Effect of different gamma-irradiation doses on cytotoxicity and material properties of porous polyether-urethane polymer.

Biomaterials respond to sterilization methods differently. Steam sterilization might decrease the performance of thermoplastic polyether-urethane (TPU); however, the effect of different gamma-radiation doses on this polymer is contradictory in present literature. The purpose of this study was to investigate the differences between irradiative doses in comparison with steam sterilization on a porous TPU scaffold produced by a new processing method. No significant differences in the surface chemical structure were found with attenuated total reflectance Fourier transform infrared spectroscopy (ATR-FTIR) analysis when comparing with the sterilization methods. The molecular weight (M(w)) had a net increase from 11.5 +/- 0.039 to 13.2 +/- 0.072 kDa by gamma-sterilization from 10 to 60 kGy. The samples that were irradiated (>60 kGy) had also an increase in polydispersity index (PDI; 1.45 +/- 0.007) in comparison with the nonsterile ones (1.31 +/- 0.017), which indicate branching. Liquid chromatography/mass spectroscopy (LC/MS) analysis showed that there was a correlation between the concentration of the breakdown product, methyl dianiline, and cytotoxicity. The concentration of this compound was found to be four times higher in steam-sterilized sample (1.3 +/- 0.01 ppb) compared with that of the polymer sample gamma-sterilized at 10 kGy (0.3 +/- 0.01 ppb). The cytotoxicity of TPU was found to decrease with higher radiation doses, and was significantly higher for the steam-sterilized samples. It is recommended that TPU produced with the described foaming method should be sterilized by gamma-irradiation at 25 kGy or higher doses.

Biocompatible Materials↗

Comparative studies on the effects of water, ethanol and water/ethanol mixtures on chemical partitioning into porcine stratum corneum and silastic membrane.

The effects of water and ethanol vehicles on stratum corneum and silastic membrane partitioning of 11 industrial and agricultural compounds were studied to aid in characterizing and assessing risk from skin exposure. Zero percent, 50% and 100% aqueous ethanol solutions were used as solvents for (14)C labeled phenol, 4-nitrophenol, pentachlorophenol, dimethyl parathion, parathion, chloropyrifos, fenthion, triazine, atrazine, simazine and propazine. Compound partitioning between the solvents and porcine stratum corneum/silastic membrane were estimated. Stratum corneum was exposed to aqueous ethanol ranging from 0% to 100% v/v ethanol in 20% increments and Fourier transform infrared spectroscopy (FT-IR) was used to obtain an index of lipid disorder. Gravimetry and FT-IR were used to demonstrate lipid extraction in aqueous ethanol solutions. Partitioning patterns in silastic membranes resembled those in stratum corneum and were correlated with octanol/water partitioning. Partitioning was highest in water and was higher from 50% ethanol than from 100% ethanol, except for parathion, 4-nitrophenol, atrazine and propazine. Correlation existed between molecular weight and partitioning in water, but not in ethanol and ethanol/water mixtures. Lipid order, as reflected in FT-IR spectra, was not altered. These studies suggest that stratum corneum partitioning of the compounds tested is primarily determined by relative compound solubility between the stratum corneum lipids and the donor solvent. Linear relationships existed between octanol/water partitioning and stratum corneum partitioning. Partitioning was also correlated with molecular weight in water solvent systems, but not in ethanol and ethanol/water mixtures. Ethanol and ethanol/water mixtures altered the stratum corneum through lipid extraction, rather than through disruption of lipid order.

Animals↗

Fourier transform infrared evidence for early deprotonation of Asp(85) at alkaline pH in the photocycle of bacteriorhodopsin mutants containing E194Q.

The role of the extracellular Glu side chains of bacteriorhodopsin in the proton transport mechanism has been studied using the single mutants E9Q, E74Q, E194Q, and E204Q; the triple mutant E9Q/E194Q/E204Q; and the quadruple mutant E9Q/E74Q/E194Q/E204Q. Steady-state difference and deconvoluted Fourier transform infrared spectroscopy has been applied to analyze the M- and N-like intermediates in membrane films maintained at a controlled humidity, at 243 and 277 K at alkaline pH. The mutants E9Q and E74Q gave spectra similar to that of wild type, whereas E194Q, E9Q/E194Q/E204Q, and E9Q/E74Q/E194Q/E204Q showed at 277 K a N-like intermediate with a single negative peak at 1742 cm(-1), indicating that Asp(85) and Asp(96) are deprotonated. Under the same conditions E204Q showed a positive peak at 1762 cm(-1) and a negative peak at 1742 cm(-1), revealing the presence of protonated Asp(85) (in an M intermediate environment) and deprotonated Asp(96). These results indicate that in E194Q-containing mutants, the second increase in the Asp(85) pK(a) is inhibited because of lack of deprotonation of the proton release group. Our data suggest that Glu(194) is the group that controls the pK(a) of Asp(85).

Amino Acid Substitution↗

A vibrational spectroscopic investigation of interactions of agonists with GluR0, a prokaryotic glutamate receptor.

We have used Fourier transform infrared spectroscopy to provide a detailed picture of the interactions between the carboxylate groups of the ligands, glutamate, serine, and glutamine, with the ligand-binding domain of a prokaryotic ionotropic glutamate receptor (GluR0). The vibrational spectra indicate that the noncovalent interactions between the 1C(alpha)-carboxylate moiety of the ligand and the protein are stronger for glutamate than for serine and glutamine. These results correlate well with the higher affinity of glutamate for GluR0-S1S2 relative to the affinities of serine and glutamine. In addition, all three ligands induce similar changes in the vibrational spectra and intrinsic fluorescence of the protein, which indicates that all three ligands induce the same structural changes in the protein. These results are consistent with the recent crystal structures of the glutamate and serine bound forms of GluR0-S1S2 and in addition provide insights into the structure of the glutamine bound form of the protein.

Bacterial Proteins↗

Effects of the sugar headgroup of a glycoglycerolipid on the phase behavior of phospholipid model membranes in the dry state.

Glycolipids are important components of almost all biological membranes. They possess unique properties that have only been incompletely characterized so far. The plant glycolipid digalactosyldiacylglycerol (DGDG) strongly influences the physical behavior of phospholipid model membranes in both the dry and hydrated state. It was, however, unclear whether the strong effect of DGDG on the gel to liquid-crystalline phase transition temperature (Tm) in dry phosphatidylcholine (PC) bilayers is mainly due to the high degree of unsaturation of the DGDG fatty acyl chains or to interactions between the DGDG and PC headgroups. Also, no information on the relative effectiveness of membrane bound and free sugars on membrane phase behavior was available. We have used Fourier-transform infrared spectroscopy (FTIR) to investigate the phase properties and H-bonding patterns in dry membranes made from 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphatidylcholine (POPC) containing one saturated and one monounsaturated (16:0/18:1) fatty acid and different fractions of DGDG or 1,2-dilinolenoyl-sn-glycero-3-phosphatidylcholine (DLPC) (18:3/18:3). This was compared to the effects of galactose (Gal) and digalactose (diGal). All additives depressed Tm of the dry membranes, but DGDG was much more effective than DLPC or Gal. diGal had a similar effect as DGDG, pointing to the sugar headgroup as the component with the strongest influence on membrane phase behavior. A combination of DLPC and diGal, which should theoretically be equivalent to DGDG, was much more effective than the galactolipid. H-bonding interactions with the P = O group of PC were also stronger for free diGal than for DGDG, indicating that the free sugar may be structurally more flexible to adopt an optimal conformation for interactions with the PC headgroup.

Carbohydrates↗

[Synthesis and property of nano-fumed silica derivative with quaternary ammonium group].

Nano-fumed silica derivative with quaternary ammonium group was synthesized and the antimicrobial activity was investigated. The nano-fumed silica derivative was investigated by Fourier-transform infrared spectroscopy (FTIR). The zeta potential and the size of the nano-fumed silica derivative were measured. The antimicrobial properties of the nano-fumed silica derivative against selected microorganisms were tested by the quantitative suspension method. The zeta potential showed that the isoelectric points of nano-fumed silica and modified nano-fumed silica are pH=4. 8 and pH =10.5-10.8, respectively, and the shift of the isoelectric point is due to the quaternary ammonium salt. The obtained nano-fumed silica derivative inhibited the growth of Gram-negative (Escherichia coli), Gram-positive bacteria (Staphylococcus aureus), and fungus (Candida albicans). The inhibiting effect of nano-fumed silica derivative on the microorganisms varied with the time of exposure. The bacteriostatic rates were noted to be 99.99%, 99.99% and 94.12% after 15 minutes' exposure, respectively. Thus the results indicate that nano-fumed silica derivative with quaternary ammonium group has significant inhibitory effect on the growth of bacteria.

Anti-Infective Agents↗

Surface characteristics and properties of lumbrokinase-immobilized polyurethane.

Potent and novel fibrinolytic enzymes (lumbrokinase [LK]) were extracted from the earthworm, Lumbricus rubellus. These enzymes were very stable and showed greater antithrombotic activity than other currently used fibrinolytic proteins. An LK fraction showing the most potent fibrinolytic activity was immobilized onto a polyurethane (PU) surface to investigate its enzymatic activity and antithrombotic activity. A methanol-extracted PU surface was coated with 3% (wt/vol) maleic anhydride methylvinyl ether copolymer (MAMEC)/tetrahydrofuran (THF) solution, and the surface was incubated in an LK solution/phosphate-buffered saline (PBS, pH 7.4). The surface properties were characterized by attenuated total reflectance-Fourier transform infrared spectroscopy (ATR-FTIR), electron spectroscopy for chemical analysis (ESCA), and dynamic contact angle. The stability of immobilized LK was determined by caseinolytic activity assay and the specificity of immobilized LK on fibrinogen/fibrin was observed by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE). The antithrombotic activity of immobilized LK was evaluated using an ex vivo rabbit A-A shunt experiment. LK immobilization was confirmed by ATR-FTIR and ESCA. Immobilized LK demonstrated stable proteolytic activity during various incubation periods. Immobilized LK proteolyzed fibrinogen and fibrin almost specifically, while it hardly hydrolyzed other plasma proteins including plasminogen and albumin. In the ex vivo A-A shunt experiment, the LK-immobilized surface significantly prolonged occlusion time over control surfaces. This is primarily due to the high thrombolytic activity of immobilized LK. In this work, a highly efficient surface modification method on the PU surface was developed, and this LK immobilization technique will be very useful in improving the blood compatibility of blood-contacting devices.

Animals↗

Mechanisms of percutaneous absorption of tamoxifen by terpenes: eugenol, D-limonene and menthone.

The effect of chemical penetration enhancers (e.g., eugenol, d-limonene and menthone in combination with 50% ethanol) on the in vitro percutaneous absorption of tamoxifen through porcine epidermis has been investigated. The above enhancers significantly increased (p<0.05) the permeability coefficient of tamoxifen in comparison with the control (50% ethanol). Fourier transform infrared spectroscopy (FT-IR) was employed to investigate the biophysical changes in the stratum corneum (SC) lipids by the enhancer(s). FT-IR results showed that the treatment of the SC with enhancers did not produce a blue shift in the asymmetric and symmetric C-H stretching peak positions. However, all of the above enhancers showed a decrease in peak heights and areas for both asymmetric and symmetric C-H stretching absorbances in comparison with the untreated SC. A decrease in peak heights and areas is a measure of lipid extraction. Partitioning of tamoxifen to powdered SC from control and enhancer solutions was also determined. FT-IR and partitioning studies reveal that the enhancement in the permeability coefficient of tamoxifen by eugenol and d-limonene is due to lipid extraction and improvement in the partitioning of the drug to the SC. However, menthone enhanced the permeability of tamoxifen by increasing extraction of the SC lipids and not by improving the partitioning of the drug to the SC.

Animals↗

Functional reconstitution, gene isolation and topology modelling of porins from Burkholderia pseudomallei and Burkholderia thailandensis.

The sequences for Omp38 from Burkholderia pseudomallei and Burkholderia thailandensis have been deposited in the DDBJ, EMBL, GenBank(R) and GSDB Nucleotide Sequence Databases under the accession numbers AY312416 and AY312417 respectively. The intracellular pathogen Burkholderia pseudomallei is the causative agent of tropical melioidosis, and Burkholderia thailandensis is a closely-related Gram-negative bacterium that does not cause serious disease. Like other bacteria, the major outer membrane (OM) porins of Burkholderia strains, Bps Omp38 and Bth Omp38 may have roles in antibiotic resistance and immunity. We purified both proteins and found them to be immunologically related, SDS-resistant, heat-sensitive trimers with M (r) of approx. 110000. In functional liposome-swelling assays, both proteins showed similar permeabilities for small sugar molecules, compatible with a pore diameter of between 1.2 and 1.6 nm. Secondary structure analysis by FTIR (Fourier-transform infrared) spectroscopy revealed almost identical spectra with predominantly beta-sheet structures, typical of bacterial porins. MALDI-TOF (matrix-assisted laser-desorption ionization-time of flight) MS and ESI/MS (electrospray ionization MS) analysis of each protein showed extensive sequence similarities to the OpcP1 porin from Burkholderia cepacia (later found to be 76.5% identical). Based on information from the incomplete B. pseudomallei genome-sequencing project, the genes encoding Omp38 were identified and amplified by PCR from B. pseudomallei and B. thailandensis genomic DNA. The nucleotide sequences are 99.7% identical, and the predicted processed proteins are 100% identical. Topology prediction and molecular modelling suggest that this newly-isolated and cloned porin is a 16-stranded beta-barrel and the external loops of the protein could be important determinants of the immune response to infection.

Amino Acid Sequence↗

A new haemocompatible phospholipid polyurethane based on hydrogenated poly(isoprene) soft segment.

A new haemocompatible phospholipid polyurethane based on hydrogenated poly(isoprene) glycol (HPIP) and 4,4'-methylendiphenyl diisocyanate (MDI) was synthesized using 2-[bis(2-hydroxyethyl)methyl-ammonio]ethylstearylphosphate (BESP) and 1,4-butanediol (BD) as chain extender. The bulk and surface characteristics of this material was investigated by differential scanning calorimetry (DSC), dynamic viscoelasticity and tensile property measurements, attenuated total reflectance-Fourier transform infrared spectroscopy (ATR-FTIR), and contact angle measurement. This polymer possessed a hydrophobic surface revealed by contact angle measurement. The haemocompatibility of this polyurethane was evaluated by platelet rich plasma (PRP) contacting studies and scanning electron microscopy (SEM) observation using medical grade poly(vinyl chloride) (PVC) as the reference. The results show that this new polyurethane had relatively lower platelet adhesion and limited shape change for the attached platelets compared to PVC. The clotting time of the materials in contact with platelet poor plasma (PPP) was 99, 75, and 62 s and in contact with PRP was more than 240, 100, and 86 s for new polyurethane, PVC, and glass, respectively. This new phospholipid polyurethane is expected to have wide applications as coating or structural material for blood-contacting medical equipment due to its outstanding haemocompatibility and excellent mechanical strength.

Biocompatible Materials↗

Self-assembly of fatty acid-alkylboladiamine salts.

Long-chain fatty acids are insoluble in aqueous solution and form crystal precipitates. It is then of particular importance to determine the physicochemical parameters allowing their dispersion in water to improve their bioavailability and their utilization as surfactants. Herein, we report a study on salt-free catanionic systems in aqueous solution made of mixtures between palmitic or stearic fatty acids and alkylboladiamines (Abd's) differing by their alkyl chain length. Phase contrast microscopy, solid-state NMR, Fourier transform infrared spectroscopy, and small-angle neutron scattering were used to characterize the phase behavior of these systems at molar ratio of fatty acid to Abd of 1 and 2. Whatever the Abd and the molar ratio, fatty acids were embedded at low temperature in a bilayer gel phase which crystallizes after a period of rest. At an equimolar ratio, the gel phases transited upon raising the temperature to an isotropic phase made of worm-like micelles except in the case of the ethylenediamine chain for which a lamellar fluid phase was observed. At a molar ratio of 2 and high temperature, fatty acids were embedded in a lamellar fluid phase which self-orients with its stacking axis perpendicular to the magnetic field. However, for a long alkylboladiamine such as spermine, worm-like micelles formed. The phase behavior at high temperature is discussed in terms of molecular volume.

Alkanes↗

Long-term changes of hydroxyapatite-coated dental implants.

There are many controversies about the long-term prognosis of hydroxyapatite (HA)-coated implants. Failure may be related to compositional and structural changes of the coating occurring during implantation. Two retrieved and two unused HA-coated blade-type implants were examined by stereomicroscopy, secondary electron microscopy, Fourier transform infrared spectroscopy, X-ray diffraction, and electron probe microanalysis. The objective was to investigate the HA morphology, composition, and structure, and to characterize the changes that occurred in the retrieved implant coatings. Retrieved implants presented partial loss of the coating, especially at the apical and mesiodistal edges. Remaining HA was thick and flattened in the cervical and central areas and gradually thinner and rougher towards the apical and mesiodistal edges. Increase of Cl and Mg, decrease of OH, and X-ray diffraction peak broadening were found in the retrieved implant coatings, in comparison with the unused implants. Morphological changes of the retrieved implants seem to depend on stress values in the surrounding bone and on implant mobility. Compositional changes and increased amount of lattice imperfections appeared in the retrieved implant coatings, as a result of ion substitutions in the apatite lattice. However, the present study could not confirm the influence of these changes on implant failure.

Coated Materials, Biocompatible↗

Adsorption behavior of methylene blue and its congeners on a stainless steel surface.

Methylene blue and its congeners as model dyes were adsorbed onto stainless steel particles at different ionic strengths, pH values, and ethanol contents, and the adsorption mechanism was investigated. A Fourier transform infrared spectroscopy (FTIR) analysis of the dyes adsorbed on the stainless steel plate was carried out to determine the orientations of the adsorbed dyes on stainless steel surface. The adsorption isotherms for all the dyes tested were approximated by a Langmuir equation (Q=Kq(m)C/(1+KC)) in most cases except under strongly basic conditions. From the ionic strength and ethanol content dependencies of the K value in the Langmuir equations, both the electrostatic and hydrophobic interactions were indicated to contribute to the adsorption of the dyes at neutral pH. By comparing the K and q(m) values for the methylene blue congeners and with the aid of the FTIR analyses, it was found that the kind of substituent groups at most positions of the polyheterocycles of methylene blue strongly affects the adsorption behavior, particularly the area occupied by an adsorbed dye molecule, the affinity for the stainless steel surface, and the orientation of the adsorbed dye molecule on the stainless steel surface.

Adsorption↗

Bioactive evaluation of 45S5 bioactive glass fibres and preliminary study of human osteoblast attachment.

Bioactive glass fibres can be used as tissue engineering scaffolds. In this investigation, the bioactive response of 45S5 glass fibres was assessed in simulated body fluid (SBF). Preliminary attachment of osteoblasts to the fibre surface was assessed, as were the fibre tensile strength and fracture toughness. Fourier transform infrared spectroscopy (FTIR) analysis revealed that hydroxyapatite (HA) was formed on the fibres' surface after 2-4 days in SBF. Raman micro-spectroscopic analysis was used to monitor development of the HA layer during immersion. A correlation was found between increase in intensity of the PO4(3-) peak near 964cm(-1) and appearance of crystalline HA (P-O bending peaks) using FTIR. Such results are encouraging for in situ bioactivity monitoring, as Raman spectra are insensitive to the presence of water, unlike FTIR. Average tensile strength of 45S5 fibres (79 microm diameter) was 340+/-140 MPa. Fracture toughness, determined using fracture surface analysis, was 0.7+/-0.1 MPa m1/2. Confocal microscopy revealed osteoblasts attached and spread along the fibres after 15-90 min culture. Scanning electron microscopy analysis showed that cells with filopodia and dorsal ruffles remained attached after 14 days in culture. These results are encouraging, as cell adhesion is an important first step prior to proliferation and differentiation.

Biocompatible Materials↗

Temperature jump-induced secondary structural change of the membrane protein bacteriorhodopsin in the premelting temperature region: a nanosecond time-resolved Fourier transform infrared study.

The secondary structural changes of the membrane protein, bacteriorhodopsin, are studied during the premelting reversible transition by using laser-induced temperature jump technique and nanosecond time-resolved Fourier transform infrared spectroscopy. The helical structural changes are triggered by using a 15 degrees C temperature jump induced from a preheated bacteriorhodopsin in D2O solution at a temperature of 72 degrees C. The structural transition from alphaII- to alphaI-helices is observed by following the change in the frequency of the amide I band from 1667 to 1651 cm-1 and the shift in the frequency of the amide II vibration from 1542 cm-1 to 1436 cm-1 upon H/D exchange. It is found that although the amide I band changes its frequency on a time scale of <100 ns, the H/D exchange shifts the frequency of the amide II band and causes a complex changes in the 1651-1600 cm-1 and 1530-1430 cm-1 frequency region on a longer time scale (>300 ns). Our result suggests that in this "premelting transition" temperature region of bacteriorhodopsin, an intrahelical conformation conversion of the alphaII to alphaI leads to the exposure of the hydrophobic region of the protein to the aqueous medium.

Bacteriorhodopsins↗

Development of a new photocurable composite resin with reduced curing shrinkage.

OBJECTIVE: This study was conducted to evaluate the potential of a novel trifunctional methacrylate as a component of a photocurable composite resin with reduced curing shrinkage. METHODS: Tris[4-(2'-hydroxy-3'-methacryloyloxypropoxy)phenyl]methane (TTEMA) was synthesized by reacting triphenylolmethane triglycidyl ether (TTE) with methacrylic acid in the presence of 4-(dimethylamino)pyridine. Photopolymerization reactivity and volumetric shrinkage of unfilled resins based on TTEMA were investigated by Fourier transform infrared spectroscopy and density measurements, respectively, and the results were compared with those for conventional dental monomers. A three-point bending test of the TTEMA-containing composite resin was carried out. RESULTS: TTEMA was easily prepared in a good yield of 89%. Unfilled resins of TTEMA and bis-GMA, each containing 40% TEGDMA, showed similar photopolymerization reactivity. TTEMA exhibited a very low photopolymerization shrinkage of 2.09%, and 3:2 TTEMA-TEGDMA unfilled resin revealed 10% lower shrinkage than a conventional bis-GMA system containing the same amount of TEGDMA. The flexural strength of a light-activated composite resin formulated with TTEMA is comparable to that of a bis-GMA composite resin under the same conditions. SIGNIFICANCE: TTEMA is promising for application as a photocurable dental monomer due to ease of synthesis, good polymerization reactivity, and relatively low curing shrinkage.

4-Aminopyridine↗

Conformational changes in aerolysin during the transition from the water-soluble protoxin to the membrane channel.

Proteolytic activation, oligomerization, and membrane insertion are three steps that precede channel formation by the bacterial toxin aerolysin. Using attenuated total reflection Fourier transform infrared spectroscopy (ATR-FTIR) and hydrogen-deuterium exchange, the structural changes associated with each step were analyzed. Our results show that activation induces a significant change in secondary structure, characterized by a decrease in random structure and an increase in beta-sheet content. We show that release of the propeptide is essential for this conformational change to occur and that changes are not restricted to the vicinity of the cleavage site but appear to propagate along the molecule. In contrast, subsequent oligomerization of the mature toxin does not involve any change in overall secondary structure but does involve a modification of the tertiary interactions. Finally, insertion of the heptameric complex into dimyristoylphosphatidylcholine vesicles also occurs without major modification of the secondary structure. Studies on the orientations of the secondary structures of the heptamer in the lipid bilayer have also been performed.

Bacterial Toxins↗

Photoinduced phenomena in layer-by-layer films of poly(allylamine hydrochloride) and Brilliant Yellow azodye.

The nanoscale interactions between adjacent layers of layer-by-layer (LBL) films from poly(allylamine hydrochloride) (PAH) and azodye Brilliant Yellow (BY) have been investigated, with the films employed for optical storage and the formation of surface-relief gratings. Using Fourier transform infrared spectroscopy, we identified interactions involving SO3- groups from BY and NH+ groups from PAH. These electrostatic interactions were responsible for the slow kinetics of writing in the optical storage experiments, due to a tendency to hinder photoisomerization and the subsequent reorientation of the azochromophores. The photoinduced birefringence did not saturate after one hour of exposure to the writing laser, whereas in azopolymer films, saturation is normally reached within a few minutes. On the other hand, the presence of such interactions prevented thermal relaxation of the chromophores after the writing laser was switched off, leading to a very stable written pattern. Moreover, the nanoscale interactions promoted mass transport for photoinscription of surface-relief gratings on PAH/BY LBL films, with the azochromophores being able to drag the inert PAH chains when undergoing the trans-cis-trans photoisomerization cycles. A low level of chromophore degradation was involved in the SRG photoinscription, which was confirmed with micro-Raman and fluorescence spectroscopies.

Allylamine↗