Near-infrared photoacoustic spectroscopy of proteins

A major problem encountered with the use of electronic spectroscopy in the analysis of biological materials in the ultraviolet, visible, and infrared region involves the limited range of the physical state of samples that can be examined. In an attempt to expand this range, photoacoustic spectra of both solid- and solution-state proteins have been obtained in the near-infrared region. Solid proteins generate detailed spectra in the region 1.0–2.6 μm, resulting primarily from hydrogenic overtone and combinational modes. Harmonics and combinations of amide group frequencies which display significant spectral complexity are observed between 1.4 and 1.7 μm, although they appear to manifest only limited conformational sensitivity. Solution spectra in D₂O are of much lower resolution. Assignments of peaks for both solution- and solid-state proteins are presented and the advantages and disadvantages of the use of near-infrared photoacoustic spectroscopy with proteins are discussed.     

The D1-173 amino acid is a structural determinant of the critical interaction between D1-Tyr161 (TyrZ) and D1-His190 in Photosystem II

The main cofactors of Photosystem II (PSII) are borne by the D1 and D2 subunits. In the thermophilic cyanobacterium Thermosynechococcus elongatus, three psbA genes encoding D1 are found in the genome. Among the 344 residues constituting the mature form of D1, there are 21 substitutions between PsbA1 and PsbA3, 31 between PsbA1 and PsbA2, and 27 between PsbA2 and PsbA3. In a previous study (Sugiura et al., J. Biol. Chem. 287 (2012), 13336-13347) we found that the oxidation kinetics and spectroscopic properties of Tyr_Z were altered in PsbA2-PSII when compared to PsbA(1/3)-PSII. The comparison of the different amino acid sequences identified the residues Cys144 and Pro173 found in PsbA1 and PsbA3, as being substituted in PsbA2 by Pro144 and Met173, and thus possible candidates accounting for the changes in the geometry and/or the environment of the Tyr_Z/His190 phenol/imidizol motif. Indeed, these amino acids are located upstream of the α-helix bearing Tyr_Z and between the two α-helices bearing Tyr_Z and its hydrogen-bonded partner, D1/His190. Here, site-directed mutants of PSII, PsbA3/Pro173Met and PsbA2/Met173Pro, were analyzed using X- and W-band EPR and UV-visible time-resolved absorption spectroscopy. The Pro173Met substitution in PsbA2-PSII versus PsbA3-PSII is shown to be the main structural determinant of the previously described functional differences between PsbA2-PSII and PsbA3-PSII. In PsbA2-PSII and PsbA3/Pro173Met-PSII, we found that the oxidation of Tyr_Z by P₆₈₀^+● was specifically slowed during the transition between S-states associated with proton release. We thus propose that the increase of the electrostatic charge of the Mn₄CaO₅ cluster in the S₂ and S₃ states could weaken the strength of the H-bond interaction between Tyr_Z^● and D1/His190 in PsbA2 versus PsbA3 and/or induce structural modification(s) of the water molecules network around Tyr_Z.     

Spectroscopic diagnosis of chronic fatigue syndrome by visible and near-infrared spectroscopy in serum samples

To investigate visible and near-infrared (Vis-NIR) spectroscopy enabling chronic fatigue syndrome (CFS) diagnosis, we subjected sera from CFS patients as well as healthy donors to Vis-NIR spectroscopy. Vis-NIR spectra in the 600-1100 nm region for sera from 77 CFS patients and 71 healthy donors were subjected to principal component analysis (PCA) and soft independent modeling of class analogy (SIMCA) to develop multivariate models to discriminate between CFS patients and healthy donors. The model was further assessed by the prediction of 99 masked other determinations (54 in the healthy group and 45 in the CFS patient group). The PCA model predicted successful discrimination of the masked samples. The SIMCA model predicted 54 of 54 (100%) healthy donors and 42 of 45 (93.3%) CFS patients of Vis-NIR spectra from masked serum samples correctly. These results suggest that Vis-NIR spectroscopy for sera combined with chemometrics analysis could provide a promising tool to objectively diagnose CFS.     

Genetic parameters of fat quality in pigs measured by near-infrared spectroscopy

Subcutaneous fat from Norwegian Landrace (n=3230) and Duroc (n=1769) pigs was sampled to investigate the sources of variation and genetic parameters of various fatty acids, fat moisture percentage and fat colour, with the lean meat percentage (LMP) also included as a trait representing the leanness of the pig. The pigs were from half-sib groups of station-tested boars included in the Norwegian pig breeding scheme. They were fed ad libitum to obtain an average of 113 kg live weight. Near-infrared spectroscopy (NIRS) was applied for prediction of the fatty acids and fat moisture percentage, and Minolta was used for the fat colour measurements. Heritabilities and genetic correlations were estimated with a multi-trait animal model using average information-restricted maximum likelihood (AI-REML) methodology. Fat from Landrace pigs had considerably more monounsaturated fatty acids, polyunsaturated fatty acids (PUFAs) and fat moisture, as well as less saturated fatty acids (SFAs) than fat from Duroc pigs. The heritability estimates (s.e. 0.03 to 0.08) for the various fatty acids were as follows: Palmitic, C16:0 (0.39 and 0.51 for Landrace and Duroc pigs, respectively); Palmitoleic, C16:1n-7 (0.41 and 0.50); Steric, C18:0 (0.46 and 0.54); Oleic, C18:1n-9 (0.67 and 0.57); Linoleic, C18:2n-6 (0.44 and 0.46); α-linolenic, C18:3n-3 (0.37 and 0.25) and n-6/n-3 ratio (0.06 and 0.01). The other fat quality traits revealed the following heritabilities: fat moisture (0.28 and 0.33), colour values in subcutaneous fat: L* (whiteness; 0.22 and 0.21), a* (redness; 0.13 and 0.24) and b* (yellowness; 0.07 and 0.17) and LMP (0.46 and 0.47). LMP showed high positive genetic correlations to PUFA (C18:2n-6 and C18:3n-3), which implies that selecting leaner pigs changes the fatty acid composition and deteriorates the quality of fat. Higher concentrations of PUFA are not beneficial as the ratio of n-6 and n-3 fatty acids becomes unfavourably high. Owing to the high genetic correlation between C18:2n-6 and C18:3n-3 and a low heritability for this ratio, the latter is difficult to change through selection. However, a small reduction in the ratio should be expected if selection aims at reducing the level of C18:2n-6. Selection for more C18:1n-9 is possible in view of the genetic parameters, which are favourable for eating quality, technological quality and human nutrition. The NIRS technology and the high heritabilities found in this study make it possible to implement fat quality traits to achieve the breeding goal in the selection of a lean pig with better fat quality.     

Sagebrush Characteristics Influencing Foraging Patterns of Pygmy Rabbits

The pygmy rabbit (Brachylagus idahoensis) is endemic to the sagebrush steppe landscapes in the western United States. Pygmy rabbits have adapted to this region by depending on big sagebrush (Artemisia tridentata) as a source of nutrition and concealment from predators. Being a central place forager, and a dietary specialist makes pygmy rabbits an ideal subject to study foraging tradeoffs. Our objectives were to determine if pygmy rabbit foraging patterns are influenced by the nutrient content of individual sagebrush, or the size and location of individual sagebrush. We first developed a near infrared spectroscopy assay that can be conducted in the field, with whole plant foliage, to determine individual sagebrush crude protein (CP) and acid detergent fiber (ADF) contents. We then measured the size, location, protein, and fiber contents of >1,500 foraged and non-foraged sagebrush plants surrounding pygmy rabbit burrows in 2 field sites in northern Utah, USA. We found significant site differences in sagebrush CP levels and significant season differences in sagebrush ADF levels; sagebrush closer to central burrows were higher in protein and lower in fiber. Pygmy rabbits preferred to forage on taller sagebrush plants that were closer to the central burrow but only marginally higher in CP (0.2%) and lower in ADF (1%). Sagebrush plants with a >50% chance of being foraged, were <5.6 m from the burrow, >0.67 m tall, >10.4% CP, and <34.95% ADF. The selection of closer, larger, and more nutrient-dense sagebrush may be influenced by pygmy rabbits' need for concealment from predators or a way to minimize foraging effort. © 2020 The Wildlife Society.     

Direct quantification of the four individual S states in Photosystem II using EPR spectroscopy

EPR spectroscopy is very useful in studies of the oxygen evolving cycle in Photosystem II and EPR signals from the CaMn₄ cluster are known in all S states except S₄. Many signals are insufficiently understood and the S₀, S₁, and S₃ states have not yet been quantifiable through their EPR signals. Recently, split EPR signals, induced by illumination at liquid helium temperatures, have been reported in the S₀, S₁, and S₃ states. These split signals provide new spectral probes to the S state chemistry. We have studied the flash power dependence of the S state turnover in Photosystem II membranes by monitoring the split S₀, split S₁, split S₃ and S₂ state multiline EPR signals. We demonstrate that quantification of the S₁, S₃ and S₀ states, using the split EPR signals, is indeed possible in samples with mixed S state composition. The amplitudes of all three split EPR signals are linearly correlated to the concentration of the respective S state. We also show that the S₁ → S₂ transition proceeds without misses following a saturating flash at 1 °C, whilst substantial misses occur in the S₂ → S₃ transition following the second flash.     

The effect of dim light at night on cerebral hemodynamic oscillations during sleep: A near-infrared spectroscopy study

Recent studies have reported that dim light at night (dLAN) is associated with risks of cardiovascular complications, such as hypertension and carotid atherosclerosis; however, little is known about the underlying mechanism. Here, we evaluated the effect of dLAN on the cerebrovascular system by analyzing cerebral hemodynamic oscillations using near-infrared spectroscopy (NIRS). Fourteen healthy male subjects underwent polysomnography coupled with cerebral NIRS. The data collected during sleep with dim light (10 lux) were compared with those collected during sleep under the control dark conditions for the sleep structure, cerebral hemodynamic oscillations, heart rate variability (HRV), and their electroencephalographic (EEG) power spectrum. Power spectral analysis was applied to oxy-hemoglobin concentrations calculated from the NIRS signal. Spectral densities over endothelial very-low-frequency oscillations (VLFOs) (0.003–0.02 Hz), neurogenic VLFOs (0.02–0.04 Hz), myogenic low-frequency oscillations (LFOs) (0.04–0.15 Hz), and total LFOs (0.003–0.15 Hz) were obtained for each sleep stage. The polysomnographic data revealed an increase in the N2 stage under the dLAN conditions. The spectral analysis of cerebral hemodynamics showed that the total LFOs increased significantly during slow-wave sleep (SWS) and decreased during rapid eye movement (REM) sleep. Specifically, endothelial (median of normalized value, 0.46 vs. 0.72, p = 0.019) and neurogenic (median, 0.58 vs. 0.84, p = 0.019) VLFOs were enhanced during SWS, whereas endothelial VLFOs (median, 1.93 vs. 1.47, p = 0.030) were attenuated during REM sleep. HRV analysis exhibited altered spectral densities during SWS induced by dLAN, including an increase in very-low-frequency and decreases in low-frequency and high-frequency ranges. In the EEG power spectral analysis, no significant difference was detected between the control and dLAN conditions. In conclusion, dLAN can disturb cerebral hemodynamics via the endothelial and autonomic systems without cortical involvement, predominantly during SWS, which might represent an underlying mechanism of the increased cerebrovascular risk associated with light exposure during sleep.     

Near-infrared mechanoluminescence sensor: A new method for on-site infrastructure detection

Near-infrared mechanoluminescence is a phenomenon that produces high penetrating near-infrared light under external stimulation. Near-infrared light coincides with the biological window, lower optical loss, and the fact that the mechanoluminescence material is a medium that converts mechanical energy into light energy. The near-infrared mechanoluminescence material has potential application prospects in the fields of biological imaging, medical diagnosis, and monitoring of building materials. In this article, we report on a perovskite-type Sr₃Sn₂O₇:Nd³⁺ near-infrared mechanoluminescence material, and its peaks locate in the first near-infrared window (800–1000 nm) and the second near-infrared window (1080, 1350 nm), respectively. Under the condition of pre-sintering with Li₂CO₃ as flux, the best sintering conditions are obtained, and the luminescence of material is in perfect agreement with the applied mechanical stress. In addition, a near-infrared mechanoluminescence sensor is proposed to solve the problem of building damage and timely maintenance.     

Benzothiazolium-based NIR AIE photosensitizers with type I and II ROS generation for efficient mitochondria-targeted photodynamic therapy

In this work, a near-infrared emissive photosensitizer of 3,3-dimethyl-N,N-diphenyl-2-(thiophen-2-yl)-3H-indol-6-amine functionlized benzothiazolium (DPITT) was developed. DPITT exhibited aggregation-induced emission effect and potent type I and II reactive oxygen species generation capacities after white light irradiation. Taking advantage of the cationic feature, DPITT penetrated the cell membrane and selectively accumulated in the mitochondria in living cells. Upon white light irradiation, the photosensitized DPITT was able to induce mitochondrial dysfunction, leading to cell death. Photosensitized DPITT was further applied to disrupt the multicellular tumour spheroids, demonstrating its potential application in inhibiting hypoxic solid tumours.