Time-resolved FTIR difference spectroscopy for the study of quinones in the A1 binding site in photosystem I: Identification of neutral state quinone bands

Infrared absorption bands associated with the neutral state of quinones in the A₁ binding site in photosystem I (PSI) have been difficult to identify in the past. This problem is addressed here, where time-resolved step-scan FTIR difference spectroscopy at 77 K has been used to study PSI with six different quinones incorporated into the A₁ binding site. (P700⁺A₁⁻ – P700A₁) and (A₁⁻ – A₁) FTIR difference spectra (DS) were obtained for PSI with the different quinones incorporated, and several double-difference spectra (DDS) were constructed from the DS. From analysis of the DS and DDS, in combination with density functional theory based vibrational frequency calculations of the quinones, the neutral state bands of the incorporated quinones are identified and assigned. For neutral PhQ in the A₁ binding site, infrared absorption bands were identified near 1665 and 1635 cm⁻¹, that are due to the C₁O and C₄O stretching vibrations of the incorporated PhQ, respectively. These assignments indicate a 30 cm⁻¹ separation between the C₁O and C₄O modes, considerably less than the ~80 cm⁻¹ found for similar modes of PhQ⁻. The C₄O mode downshifts due to hydrogen bonding, so the suggestion is that hydrogen bonding is weaker for the neutral state compared to the anion state, indicating radical-induced proton dynamics associated with the quinone in the A₁ binding site in PSI.     

In vitro cytotoxicity and pro-apoptotic activity of phycocyanin nanoparticles from Ulva lactuca (Chlorophyta) algae

This study investigated the in vitro antioxidant, proapoptotic and anti-proliferative activity of phycocyanin extracted from Ulva lactuca (Chlorophyta) algae extract loaded on albumin nanoparticle (ULANP). The characterization of ULANP profile was done by using FTIR and its cytotoxicity was investigated by using MTT assay against HepG2 and MCF7 cell lines. The proapoptotic markers caspase 8 & 9 were measured. Analysis of ULANP by FTIR showed the characteristic band (2100 cm⁻¹ ~3700 cm⁻¹) that is indicated primarily by COO, CO and conjugated double bond. These bonds showed the spectral band at peaks of 2985 cm⁻¹ and 2860 cm⁻¹, 2986 cm⁻¹ respectively. The antioxidant potential and radical scavenging property of ULANP was also appreciable as compared to the vitamin C and gallic acid. The antiproliferative assay carried out by WST-1 suggests that ULANP was effective against both HepG2 (93.17%) and MCF7 (91.3%). Caspase-8 and −9 were significantly elevated (p < 0.001) in both the cell lines of breast and liver cancer. It was concluded that ULANP induced anti-proliferative and pro-apoptotic activities on liver and breast cancer. It is promising as a novel antitumor activity for further investigation the mechanistic pathways mediated this action.     

New insights into hidradenitis suppurativa diagnosis via salivary infrared biosignatures: A pilot study

Hidradenitis suppurativa (HS) is a chronic inflammatory skin disease which can lead to a prolonged physical disability. HS diagnosis is exclusively clinical with the absence of biomarkers. Our study aims at assessing the HS-diagnostic potential of infrared spectroscopy from saliva, as a biofluid reflecting the body's pathophysiological state. Infrared spectra from 127 patients (57 HS and 70 non-HS) were processed by multivariate methods: principal component analysis coupled with Kruskal–Wallis or Mann–Whitney tests to identify discriminant spectral wavenumbers and linear discriminant analysis to evaluate the performances of HS-diagnostic approach. Infrared features, mainly in the 1300 cm⁻¹-1600 cm⁻¹ region, were identified as discriminant for HS and prediction models revealed diagnostic performances of about 80%. Tobacco and obesity, two main HS risk factors, do not seem to alter the infrared diagnosis. This pilot study shows the potential of salivary “liquid biopsy” associated to vibrational spectroscopy to develop a personalized medical approach for HS patients' management.     

Raman spectroscopy of human skin for kidney failure detection

The object of this paper is in vivo study of skin spectral-characteristics in patients with kidney failure by conventional Raman spectroscopy in near infrared region. The experimental dataset was subjected to discriminant analysis with the projection on latent structures (PLS-DA). Application of Raman spectroscopy to investigate the forearm skin in 85 adult patients with kidney failure (90 spectra) and 40 healthy adult volunteers (80 spectra) has yielded the accuracy of 0.96, sensitivity of 0.94 and specificity of 0.99 in terms of identifying the target subjects with kidney failure. The autofluorescence analysis in the near infrared region identified the patients with kidney failure among healthy volunteers of the same age group with specificity, sensitivity, and accuracy of 0.91, 0.84, and 0.88, respectively. When classifying subjects by the presence of kidney failure using the PLS-DA method, the most informative Raman spectral bands are 1315 to 1330, 1450 to 1460, 1700 to 1800 cm⁻¹. In general, the performed study demonstrates that for in vivo skin analysis, the conventional Raman spectroscopy can provide the basis for cost-effective and accurate detection of kidney failure and associated metabolic changes in the skin.     

FT-IR spectroscopic evidence of sugar ring conformational changes in GpC and CpG on platination and intercalation

An FT-IR spectroscopic study concerning changes in the conformation of sugar in the dinucleotides; GpC and CpG, on platination and intercalation is presented. The results are compared with the FT-IR spectral data of 5′-CMP, 5′-GMP, 3′-GMP and their metal adducts. The spectra of free GpC, free CpG, proflavine-GpC, proflavine-CpG, and cis-[Pt(NH₃)₂(GpC)₂]²⁺ exhibit the diagnostic band at 800 cm⁻¹ which was assigned to a sugar phosphate vibrational mode and diagnostic of C3′-endo sugar pucker. In the case of 9-aminoacridine-GpC and cis-[Pt(NH₃)₂(CpG]⁺ the diagnostic bands of the C2′-endo and C3′-endo conformations are observed at 810–820 cm⁻¹ and near 800 cm⁻¹ respectively. The results are in good agreement with X-ray data. The infrared diagnostic bands are important for distinguishing the sugar pucker conformational changes.     

Spectroscopic studies of pig kidney diamine oxidase-anion complexes

Complexes of pig kidney diamine oxidase with azide, thiocyanate, and cyanide have been characterized by EPR and circular dichroism spectroscopy. Cu(II) d-d bands are observed in the CD spectrum of the resting enzyme at ≈800 nm (12 500 cm⁻¹) and 575 nm (17 400 cm⁻¹). Anion binding produces characteristic changes in the CD spectra. N₃⁻/SCN⁻ → Cu(II) ligand-to-metal charge-transfer transitions are located at 390 nm (25 600 cm⁻¹) and 365 nm (27 400 cm⁻¹), respectively. In addition, an intense new band grew in at ≈500 nm (20 000 cm⁻¹) when azide or thiocyanate were added, which may be assigned as a Cu(II) electronic transition that gains rotational strength in the anion complex. EPR spectra established that the Cu(II)-anion complexes are tetragonal; however, the magnitudes of the anion-induced shifts in the EPR parameters were consistent with substantial perturbations of the Cu(II) electronic ground state in the thiocyanate and cyanide complexes. Prominent superhyperfine splitting was apparent in the EPR spectra of the diamine oxidase complexes with thiocyanate and cyanide. The superhyperfine structure is (at least) partially attributable to endogenous Cu(II) ligands, probably from imidazole.     

Studying the genotoxic effects of high intensity terahertz radiation on fibroblasts and CNS tumor cells

The data is obtained on the effect of high-intensity pulses of terahertz (THz) radiation with a broad spectrum (0.2–3 THz) on cell cultures. We have evaluated the threshold exposure parameters of THz radiation causing genotoxic effects in fibroblasts. Phosphorylation of histone H2AX at Ser 139 (γH2AX) was chosen as a marker for genotoxicity and a quantitative estimation of γH2AX foci number in fibroblasts was performed after cell irradiation with THz pulses for 30 min. No genotoxic effects of THz radiation were observed in fibroblasts unless peak intensity and electric field strength exceeded 21 GW cm⁻² and 2.8 MV cm⁻¹, respectively. In tumor cell lines (neuroblastoma (SK-N-BE (2)) and glioblastoma (U87)), exposure to THz pulses with peak intensity of 21 GW cm⁻² for 30 min caused no morphological changes as well as no statistically significant increase in histone phosphorylation foci number.     

Raman spectroscopic analysis of the sodium salt of kappa-carrageenan and related compounds in solution

Laser-Raman spectra of Na⁺ kappa-carrageenan, Na⁺ neocarrabiose 4-sulphate, and neocarrabiose in the region 700–1500 cm⁻¹ are reported for solutions in H₂O and D₂O. The C-1-H-1α vibration, coupled with COH related modes, is assigned to a band at 840 cm⁻¹, close to the maximum of the symmetrical COS stretching (∼850 cm⁻¹). The symmetrical SO stretch is proposed to occur near 1040 cm⁻¹ and is probably coupled with COH vibrations which give rise to strong bands in the region 1000–1100 cm⁻¹. The intense band in the region 730–740 cm⁻¹ is ascribed to a complex ring vibration.     

EPR,ligand field spectra and antimicrobial activity of dimeric aryloxyacetatocopper(II) complexes with antipyrine

Copper(lI) complexes with aryloxyacetato ligands and antipyrine (Apy) of the general formula [Cu₂- (RCOO)₄(Apy)₂] were prepared with RCOO⁻ as phenoxyacetato, 3-methylphenoxyacetato, 4-methylphenoxyacetato, 4-chlorophenoxyacetato, 2-methyl- 4-chlorophenoxyacetato, 3-methyl-4-chlorophenoxyacetato and 2-naphthoxyacetato anions. The parameters of the triplet spin state EPR spectra at X and Q band frequencies (D|hc = 0.37 cm⁻¹, g⊥ = 2.07, g6 = 2.38, A6|c = 0.007 cm⁻¹) provide evidence of a dimeric carboxylato-bridged structure with apically bound antipyrine. The almost constant EPR data as well as the energies of the ligand field transitions at 77 K [d_x²−y² ← d_xz, d_yz (14000 cm⁻¹), d_x²−y² ← d_xy (11200 cm⁻¹) and d_x²−y² ← d_z² (9500 cm⁻¹)] indicate that the square-pyramidal geometry of the CuO₅ polyhedra is nearly unaffected by the substituents on the aryloxyacetato group. The complexes show antimicrobial activity, being most efficient against Candida albicans and Bacillus subtilis.     

Weak Electron Phonon Coupling and Deep Level Impurity for High Thermoelectric Performance Pb1−xGaxTe

High ZT of 1.34 at 766 K and a record high average ZT above 1 in the temperature range of 300‐864 K are attained in n‐type PbTe by engineering the temperature‐dependent carrier concentration and weakening electron–phonon coupling upon Ga doping. The experimental studies and first principles band structure calculations show that doping with Ga introduces a shallow level impurity contributing extrinsic carriers and imparts a deeper impurity level that ionizes at higher temperatures. This adjusts the carrier concentration closer to the temperature‐dependent optimum and thus maximizes the power factor in a wide temperature range. The maximum power factor of 35 µW cm⁻¹ K⁻² is achieved for the Pb_0.98Ga_0.02Te compound, and is maintained over 20 µWcm⁻¹ K⁻² from 300 to 767 K. Band structure calculations and X‐ray photoelectron spectroscopy corroborate the amphoteric role of Ga in PbTe as the origin of shallow and deep levels. Additionally, Ga doping weakens the electron–phonon coupling, leading to high carrier mobilities in excess of 1200 cm² V⁻¹ s⁻¹. Enhanced point defect phonon scattering yields a reduced lattice thermal conductivity. This work provides a new avenue, beyond the conventional shallow level doping, for further improving the average ZT in thermoelectric materials.     

Toxicity mechanism of Cu2+ ion individually and in combination with Zn2+ ion in characterizing the molecular changes of Staphylococcus aureus studied using FTIR coupled with chemometric analysis

Copper and zinc have a high binding affinity with a Staphylococcus aureus bacterial community. This causes a change in the biomolecular composition of S. aureus. Our study aims at understanding the resistance mechanism of Cu and Zn either or in various combinations using FTIR and chemometric techniques. Zn toxicity resulted in a significant change in lipid content (3100-2800 cm⁻¹) compared to Cu. A significant decrease in protein content is observed for Cu treatment in the amide region. The bio-concentration factor shows a higher value for Cu compared to Zn. The increase in band area of carbohydrates moieties 1059 cm⁻¹ shows the secretion of EPS due to Cu toxicity. A significant change in nucleic acid compositions was noted in the region1200-900 cm⁻¹ due to Zn treatment. Secondary structural change in protein shows β sheet formation. The result of the finding shows Cu has greater toxicity than Zn. Further toxicity effects were greatly enhanced for metal mixtures ratio (Cu:2Zn). This shows Zn exhibits synergism effect with Cu. The obtained ROC (receiver operating characteristic) curve area gives good reliability of the experiments. The study attempts to understand the mechanism of toxicity removal of Cu and Zn metal mixtures by bacterial population using FTIR coupled with chemometric techniques. Open in a separate windowGraphical abstract     

On transverse ordering in biomembranes by Raman spectroscopy

Computer simulation analysis of the structure sensitive features in the Raman spectrum of aqueous dipalmitoyl phosphatidylcholine (DPPC) multilayers was made for the 1000–1200 cm₋₁ region. The composite triplet was resolved into 4 Lorentzian bands with the possibility to follow their parameters (amplitude, width and position) in the temperature interval 25–60°C. The analysis reveals the apparent inward shift of the 1130 and 1065 cm⁻¹ bands towards both sides of the broad intense 1087 cm⁻¹ feature which is due to the relative changes in amplitudes of these bands in the course of the lipid phase transition. The more accurate method for the evaluation of the trans order parameter (S_trans) confirms the underestimation of the trans segment content above the phase transition temperature when using both the relative amplitude and area of the 1130 cm⁻¹ band for the quantitative characterization of chain conformation. Calculated changes in the bilayer thickness following the frequency shift of the 1100 cm⁻¹ band are in good agreement with the previously reported deuterium NMR data and X-ray diffraction studies.     

Raman intensities of carbon-carbon stretching modes in a model membrane

Laser-Raman spectra of L-α-dimyristoylphosphatidylcholine (DMPC) liposomes in the spectral range 1000–1200 cm^?1 were obtained as a function of temperature from ?80 to +50°C. The triplet found in this spectral region was resolved into Lorentzian components by means of an iterative computer program. The peak intensities, band widths, and band areas of the resolved 1062 cm^?1 and 1130 cm^?1 bands, assigned to CC stretching vibrations of trans segments, were evaluated as a function of temperature. While the peak intensities of the bands decrease substantially with temperature, the band widths show a considerable increase. The change in band areas is therefore smaller than the change in peak heights. Experiments with all trans carboxylic acids showed that in these compounds the area of the Raman bands at 1062 cm^?1 and 1130 cm^?1 is proportional to the number of trans bonds. The variation with temperature of the number of trans and gauche bonds in the studied phospholipid is reflected by the change of the area of the 1130 cm^?1 Raman band.     

Sequence dependent DNA conformations: Raman spectroscopic studies and a model of action of restriction enzymes

Raman spectra have been examined to clarify the polymorphic forms of DNA, A, B, and Z forms. From an analysis we found that the guanine ring breathing vibration is sensitive to its local conformation. Examination of nine crystals of guanosine residues in which the local conformations are well established revealed that a guanosine residue with a C3′endo-anti gives a strong line at 666±2cm⁻¹, O4′endo-anti at 682 cm⁻¹, Cl′exo-anti at 673 cm⁻¹, C2′endo-anti at 677 cm⁻¹ and syn-forms around 625 cm⁻¹. Using this characteristic line, we were able to obtain the local conformations of guanosine moieties in poly(dG-dC).Such a sequence derived variation is suggested to be recognized by sequence specific proteins such as restriction enzymes. We found a correlation between sequence dependent DNA conformation and a mode of action of restriction enzymes. The cutting mode of restriction enzymes is classified into three groups. The classification of whether the products have blunt ends, two-base-long cohesive ends, or four-base-long cohesive ends depends primarily on the substrate, not on the enzyme. It is suggested that sequence dependent DNA conformation causes such a classification by the use of the Calladine-Dickerson analysis. In the recognition of restriction enzymes, the methyl group in a certain sequence is considered to play an important role by changing the local conformation of DNA.     

Responses to light by a nonphototactic mutant ofDictyostelium discoideum

The responses of the nonphototactic mutant P73 ofDictyostelium discoideum to light stimuli have been investigated and compared with those of the axenin strain Ax2. (1) Pseudoplasmodia of the mutant P73 show no phototactic response to unilateral white light in the range from 2 μW · cm⁻² to 2 × 10⁴ μW · cm⁻². Mixing experiments demonstrated that pseudoplasmodia composed of as little as 10% wild-type amoebas and 90% P73 amoebas showed almost normal phototaxis. (2) Amoebas of the mutant P73 accumulated in light traps of intensities between 10⁴ and 8 × 10⁴ μW · cm⁻² and dispersed from a trap of 10⁵ μW · cm⁻². The sensitivity of both responses was about a factor of 10 lower than that in Ax2. (3) White light inhibited aggregation of P73 amoebas in the range from 10³ to 10⁵ μW · cm⁻², about a factor of 10 higher than that in Ax2.     

Quantification of perfusion and metabolism in an autism mouse model assessed by diffuse correlation spectroscopy and near-infrared spectroscopy

There is a need for quantitative biomarkers for early diagnosis of autism. Cerebral blood flow and oxidative metabolism parameters may show superior contrasts for improved characterization. Diffuse correlation spectroscopy (DCS) has been shown to be reliable method to obtain cerebral blood flow contrast in animals and humans. Thus, in this study, we evaluated the combination of DCS and fNIRS in an established autism mouse model. Our results indicate that autistic group had significantly (P = .001) lower (~40%) blood flow (1.16 ± 0.26) × 10⁻⁸ cm²/s), and significantly (P = .015) lower (~70%) oxidative metabolism (52.4 ± 16.6 μmol/100 g/min) compared to control group ([1.93 ± 0.74] × 10⁻⁸ cm²/s, 177.2 ± 45.8 μmol/100 g/min, respectively). These results suggest that the combination of DCS and fNIRS can provide hemodynamic and metabolic contrasts for in vivo assessment of autism pathological conditions noninvasively.     

1H- and 31P-NMR studies on smooth muscle of bullfrog stomach

³¹P-NMR spectra of bullfrog stomach smooth muscle showed peaks for creatine phosphate (4.8 μmol·g⁻¹ wet wt.), ATP (3.6), inorganic phosphate (P_i, 2.4), phosphomonoesters (3.0) and phosphodiesters (3.3). The intracellular pH was 7.3, and calculated from the chemical shift of P_i. ¹H-NMR spectra of smooth muscle yielded peaks of 2.9 for lactate, 6.6 for total creatine (creatine phosphate + creatine) and methyl protons of choline tentatively assigned to glycerolphosphorylcholine or to membrane phospholipids. Creatine phosphate and ATP decreased under anaerobic conditions, and intracellular acidification was observed with the concomitant increase in lactate. ³¹P saturation transfer studies showed that saturation of the γ-ATP resonance reduced the intensity of creatine phosphate to 60% of its control value, and the measured T₁ value of creatine phosphate was 2.4 s with saturation. The calculated forward flux of the creatine kinase reaction (decomposition direction of creatine phosphate) was 0.77 μmol·g⁻¹ wet wt.·s⁻¹. The creatine kinase flux was approx. 100-times larger than the ATP turnover rate, calculated from the oxygen consumption rate with the assumption, P/O = 3. In conclusion, the creatine kinase reaction is at equilibrium in resting smooth muscle of bullfrog stomach.     

High within-stream replication is needed to predict litter fluxes in wet–dry tropical streams

1. Streams draining forested landscapes are fuelled by terrestrial plant litter, which can be transported downstream or retained and broken down locally. However, fluxes of plant litter in streams can vary at multiple spatio-temporal scales, affecting the availability of this key resource in heterotrophic stream food webs.
2. To explore this question we quantified several processes related to litter dynamics (i.e. litter inputs, storage, losses by transport and losses by breakdown) by sampling litter at multiple sites in three streams of the Brazilian Cerrado biome (which has a tropical wet–dry climate) for 2 years. We assessed the relative contribution of different spatial (among and within streams) and temporal scales (annual, seasonal and monthly) to total variability of these processes (hereafter fluxes).
3. Spatial and temporal variability of fluxes were both high, but spatial variation was 1.67-fold greater than temporal variation (61 versus 37%, respectively), especially at the within-stream scale (50% overall); an exception was litterfall, which varied less spatially than temporally (24 versus 76%). Temporal variation of litter storage (and hence availability to consumers) was mostly seasonal and due to differences in net transport.
4. Inputs and transport were higher in the wet than the dry season (wet versus dry season, 1.45 versus 0.92 and 1.43 versus 0.06 g litter m⁻² day⁻¹), while breakdown was similar between both seasons (0.88 versus 0.94 g litter m⁻² day⁻¹). Storage (i.e. accumulation) rate was positive and negative in the dry and wet season, respectively, indicating that litter was stored in the dry season and exported in the wet season. The transitional dry–wet season showed the highest inputs, breakdown and storage (3.21, 1.63 g litter m⁻² day⁻¹ and 145 g litter m⁻²), while the wet–dry season showed lower inputs (as in the dry season), higher transport (as in the wet season) and lower breakdown and storage than the other seasons (0.93, 0.65, 0.31 g litter m⁻² day⁻¹ and 24 g litter m⁻²).
5. Our results underscore the role of variation in biophysical drivers of litter fluxes within streams (e.g. pool–riffle configuration, substrate features, biological communities), and suggest that high within-stream replication is necessary to study litter fluxes at larger scales and over time. The seasonal patterns suggested potential changes in litter dynamics under future climate scenarios in the tropics, including increased storage due to reduced transport in a drier climate.

     

The effect of tube focal spot size and acquisition mode on task-based image quality performance of a GE revolution HD dual energy CT scanner

PurposeTo assess the task-based performance of images obtained under different focal spot size and acquisition mode on a dual-energy CT scanner.MethodsAxial CT image series of the Catphan phantom were obtained using a tube focus at different sizes. Acquisitions were performed in standard single-energy, high resolution (HR) and dual-energy modes. Images were reconstructed using conventional and high definition (HD) kernels. Task-based transfer function at the 50% level (TTF_50%) for teflon, delrin, low density polyethylene (LDPE) and acrylic, as well as image noise and noise texture, were assessed across all focal spots and acquisition modes using Noise Power Spectrum (NPS) analysis. A non-prewhitening mathematical observer model was used to calculate detectability index ($d_{\mathit{NPW}}^{\prime }$).ResultsTTF_50% degraded with increasing focal spot size. TTF_50% ranged from 0.67 mm⁻¹ for teflon to 0.25 mm⁻¹ for acrylic. For standard kernel, image noise and NPS-determined average spatial frequency were 8.3 HU and 0.29 mm⁻¹, respectively in single-energy, 12.0 HU and 0.37 mm⁻¹ in HR, and 7.9 HU and 0.26 mm⁻¹ in dual-energy mode. For standard kernel, $d_{\mathit{NPW}}^{\prime }$ was 61 in single-energy and HR mode and reduced to 56 in dual-energy mode.ConclusionsThe task-based image quality assessment metrics have shown that spatial resolution is higher for higher image contrast materials and detectability is higher in the standard single-energy mode compared to HR and dual-energy mode. The results of the current study provide CT operators the required knowledge to characterize their CT system towards the optimization of its clinical performance.     

A computationally efficient Monte-Carlo model for biomedical Raman spectroscopy

Monte Carlo (MC) modeling is a valuable tool to gain fundamental understanding of light-tissue interactions, provide guidance and assessment to optical instrument designs, and help analyze experimental data. It has been a major challenge to efficiently extend MC towards modeling of bulk-tissue Raman spectroscopy (RS) due to the wide spectral range, relatively sharp spectral features, and presence of background autofluorescence. Here, we report a computationally efficient MC approach for RS by adapting the massively-parallel Monte Carlo eXtreme (MCX) simulator. Simulation efficiency is achieved through “isoweight,” a novel approach that combines the statistical generation of Raman scattered and Fluorescence emission with a lookup-table-based technique well-suited for parallelization. The MC model uses a graphics processor to produce dense Raman and fluorescence spectra over a range of 800 − 2000 cm⁻¹ with an approximately 100× increase in speed over prior RS Monte Carlo methods. The simulated RS signals are compared against experimentally collected spectra from gelatin phantoms, showing a strong correlation.