Europium-151 and iron-57 nuclear resonant vibrational spectroscopy of naturally abundant KEu(III)Fe(II)(CN)6 and Eu(III)Fe(III)(CN)6 complexes.

We have performed and analyzed the first combined 151Eu and 57Fe nuclear resonant vibrational spectroscopy (NRVS) for naturally abundant KEu(III)[Fe(II)(CN)6] and Eu(III)[Fe(III)(CN)6] complexes. Comparison of the observed 151Eu vs.57Fe NRVS spectroscopic features confirms that Eu(III) in both KEu(III)[Fe(II)(CN)6] and Eu(III)[Fe(III)(CN)6] occupies a position outside the [Fe(CN)6] core and coordinates to the N atoms of the CN- ions, whereas Fe(III) or Fe(II) occupies the site inside the [Fe(CN)6]4- core and coordinates to the C atoms of the CN- ions. In addition to the spectroscopic interest, the results from this study provide invaluable insights for the design and evaluation of the nanoparticles of such complexes as potential cellular contrast agents for their use in magnetic resonance imaging. The combined 151Eu and 57Fe NRVS measurements are also among the first few explorations of bi-isotopic NRVS experiments.

Nitrogenase Chemistry at 10 Kelvin─Phototautomerization and Recombination of CO-Inhibited α-H195Q Enzyme.

CO-bound forms of nitrogenase are N2-reduction inhibited and likely intermediates in Fischer-Tropsch chemistry. Visible-light photolysis at 7 K was used to interrogate all three known CO-related EPR-active forms as exhibited by the α-H195Q variant of Azotobacter vinelandii nitrogenase MoFe protein. The hi(5)-CO EPR signal converted to the hi-CO EPR signal, which reverted at 10 K. FT-IR monitoring revealed an exquisitely light-sensitive "Hi-2" species with bands at 1932 and 1866 cm-1 that yielded "Hi-1" with bands at 1969 and 1692 cm-1, which reverted to "Hi-2". The similarities of photochemical behavior and recombination kinetics showed, for the first time, that hi-CO EPR and "Hi-1" IR signals arise from one chemical species. hi(5)-CO EPR and "Hi-2" IR signals are from a second species, and lo-CO EPR and "Lo-2" IR signals, formed after prolonged illumination, are from a third species. Comparing FT-IR data with CO-inhibited MoFe-protein crystal structures allowed assignment of CO-bonding geometries in these species.

A preoperative radiomics model for the identification of lymph node metastasis in patients with early-stage cervical squamous cell carcinoma.

OBJECTIVES: To develop and validate a radiomics model for preoperative identification of lymph node metastasis (LNM) in patients with early-stage cervical squamous cell carcinoma (CSCC). METHODS: Total of 190 eligible patients were randomly divided into training (n = 100) and validation (n = 90) cohorts. Handcrafted features and deep-learning features were extracted from T2W fat suppression images. The minimum redundancy maximum relevance algorithm and LASSO regression with 10-fold cross-validation were used for key features selection. A radiomics model that incorporated the handcrafted-signature, deep-signature, and squamous cell carcinoma antigen (SCC-Ag) levels was developed by logistic regression. The model performance was assessed and validated with respect to its calibration, discrimination, and clinical usefulness. RESULTS: Three handcrafted features and three deep-learning features were selected and used to build handcrafted- and deep-signature. The model, which incorporated the handcrafted-signature, deep-signature, and SCC-Ag, showed satisfactory calibration and discrimination in the training cohort (AUC: 0.852, 95% CI: 0.761-0.943) and the validation cohort (AUC: 0.815, 95% CI: 0.711-0.919). Decision curve analysis indicated the clinical usefulness of the radiomics model. The radiomics model yielded greater AUCs than either the radiomics signature (AUC = 0.806 and 0.779, respectively) or the SCC-Ag (AUC = 0.735 and 0.688, respectively) alone in both the training and validation cohorts. CONCLUSION: The presented radiomics model can be used for preoperative identification of LNM in patients with early-stage CSCC. Its performance outperforms that of SCC-Ag level analysis alone. ADVANCES IN KNOWLEDGE: A radiomics model incorporated radiomics signature and SCC-Ag levels demonstrated good performance in identifying LNM in patients with early-stage CSCC.

Influence of tube voltage, tube current and newer iterative reconstruction algorithms in CT perfusion imaging in rabbit liver VX2 tumors.

PURPOSE: We aimed to explore the influence of tube voltage, current and iterative reconstruction (IR) in computed tomography perfusion imaging (CTPI) and to compare CTPI parameters with microvessel density (MVD). METHODS: Hepatic CTPI with three CTPI protocols (protocol A, tube voltage/current 80 kV/40 mAs; protocol B, tube voltage/current 80 kV/80 mAs; protocol C: tube voltage/current 100 kV/80 mAs) were performed in 25 rabbit liver VX2 tumor models, and filtered back projection (FBP) and IR were used for reconstruction of raw data. Hepatic arterial perfusion (HAP), hepatic portal perfusion (HPP), total perfusion (TP), hepatic arterial perfusion index (HPI), blood flow (BF) and blood volume (BV) of VX2 tumor and normal hepatic parenchyma were measured. Image noise, signal-to-noise ratio (SNR) and contrast-to-noise ratio (CNR) were quantified and radiation dose was recorded. MVD was counted using CD34 stain and compared with CTPI parameters. RESULTS: The highest radiation dose was found in protocol C, followed by protocols B and A. IR lowered image noise and improved SNR and CNR in all three protocols. There was no statistical difference between HAP, HPP, TP, HPI, BF and BV of VX2 tumor and normal hepatic parenchyma among the three protocols (P > 0.05) with FBP or IR reconstruction, and no statistical difference between IR and FBP reconstruction (P > 0.05) in either protocol. MVD had a positive linear correlation with HAP, TP, BF, with best correlation observed with HAP; MVD of VX2 tumor showed no or poor correlation with HPI and BV. CONCLUSION: CTPI parameters are not affected by tube voltage, current or reconstruction algorithm; HAP can best reflect MVD, but no correlation exists between BV and MVD.

Unraveling the Mechanism of Catalytic Water Oxidation via de Novo Synthesis of Reactive Intermediate.

Artificial photosynthesis could promise abundant, carbon-neutral energy, but implementation is currently limited by a lack of control over the multi-electron catalysis of water oxidation. Discoveries of the most active catalysts still rely heavily on serendipity. [Ru(tpy)(bpy)(H2O)]2+ (1; bpy = 2,2'-bipyridine, tpy = 2,2';6',2″-terpyridine) is representative of the largest known class of water oxidation catalysts. We undertook an extensive spectroscopic analysis of the prototypical 1 water oxidation catalyst and its fastest known analog [Ru(EtO-tpy)(bpy)(H2O)]2+ (2), capable of 10 times faster water oxidation, to investigate the mechanism of action and factors controlling catalytic activity. EPR and resonance Raman spectroscopy did not detect the proposed [RuV═O] intermediate in 1 and 2 but indicated the possible formation of N-oxides. A lag phase was observed prior to O2 evolution, suggesting catalyst modification before the onset of catalysis. The reactive intermediate [Ru(tpy)(bpy-NO)(H2O)]2+ (1-NO; bpy-NO = 2,2'-bipyridine-N-oxide) proposed by combined spectroscopic and DFT analysis was de novo synthesized and demonstrated 100-fold greater catalytic activity than 1. Thus, in situ transient formation of small amounts of the Ru complex with N-oxide ligands can significantly activate single-site Ru-based catalysts. Furthermore, the rate of O2 evolution was found to correlate with the redox potential of the ligand. This observation might assist with rational design of new catalysts.

Radiomics nomogram outperforms size criteria in discriminating lymph node metastasis in resectable esophageal squamous cell carcinoma.

OBJECTIVES: To determine the value of radiomics in predicting lymph node (LN) metastasis in resectable esophageal squamous cell carcinoma (ESCC) patients. METHODS: Data of 230 consecutive patients were retrospectively analyzed (154 in the training set and 76 in the test set). A total of 1576 radiomics features were extracted from arterial-phase CT images of the whole primary tumor. LASSO logistic regression was performed to choose the key features and construct a radiomics signature. A radiomics nomogram incorporating this signature was developed on the basis of multivariable analysis in the training set. Nomogram performance was determined and validated with respect to its discrimination, calibration and reclassification. Clinical usefulness was estimated by decision curve analysis. RESULTS: The radiomics signature including five features was significantly associated with LN metastasis. The radiomics nomogram, which incorporated the signature and CT-reported LN status (i.e. size criteria), distinguished LN metastasis with an area under curve (AUC) of 0.758 in the training set, and performance was similar in the test set (AUC 0.773). Discrimination of the radiomics nomogram exceeded that of size criteria alone in both the training set (p <0.001) and the test set (p=0.005). Integrated discrimination improvement (IDI) and categorical net reclassification improvement (NRI) showed significant improvement in prognostic value when the radiomics signature was added to size criteria in the test set (IDI 17.3%; p<0.001; categorical NRI 52.3%; p<0.001). Decision curve analysis supported that the radiomics nomogram is superior to size criteria. CONCLUSIONS: The radiomics nomogram provides individualized risk estimation of LN metastasis in ESCC patients and outperforms size criteria. KEY POINTS: • A radiomics nomogram was built and validated to predict LN metastasis in resectable ESCC. • The radiomics nomogram outperformed size criteria. • Radiomics helps to unravel intratumor heterogeneity and can serve as a novel biomarker for determination of LN status in resectable ESCC.

The evaluation of coronary artery-to-pulmonary artery fistula in adulthood on 256-slice CT coronary angiography: Comparison with coronary catheter angiography and transthoracic echocardiography.

OBJECTIVE: To review the imaging features of coronary artery-to-pulmonary artery fistula (CPAF) on CT coronary angiography (CTCA) and evaluate its diagnostic performance compared with coronary catheter angiography (CCA) and transthoracic echocardiography (TTE). MATERIALS AND METHODS: We retrospectively reviewed with a diagnosis of CPAF from among 19855 consecutive CCTA performed with 256-slice MDCT scanner for suspected coronary artery disease. CT images were evaluated for - origin, number, size and course (tubular/worm-like dilation/significant aneurysm formation/wall attachment sign) of fistula vessels, drainage site, drainage site imaging features (pierced sign, isodensity sign, smoke sign, jet sign), and main pulmonary artery (MPA) enlargement. 25 patients of CPAF also underwent CCA and 47 patients underwent TTE. RESULTS: There were 72 patients with CPAF (0.36%) in our study, of which 44 were men and 28 were women, with mean age of 55.8 ±â€¯13.2 years (range 22-85 years). CPAF originated from conus artery, left anterior descending artery (LAD), combined conus artery and LAD in 55, 67, 50 cases, respectively. Tubular dilation, worm-like dilation and aneurysm was seen in 14, 58 and 35 cases, respectively. Wall attachment sign was noted in 69 cases. All the cases demonstrated only a single drainage site, with left lateral wall, left anterolateral, anterior, right lateral and right anterolateral walls of MPA in 44, 21, 5, 1 and 1 cases, respectively. The mean diameter of the fistula drainage site was 2.6 ±â€¯1.3 mm. Pierced sign, jet sign, smoke sign, isodensity sign was seen in 72, 46, 41 and 24 cases, respectively. MPA enlargement was seen in 20 patients. CCA showed CPAF in only 20 cases among 25 patients; while TTE showed CPAF in only 9 patients among 47 patients. CONCLUSION: CTCA is competent in detecting and characterizing CPAF with an excellent diagnostic performance as the first imaging modality of choice, which is valuable for giving a distinct and intuitive explanation to patients and physicians and making an objective and exact assessment for further management.

Radiomics approach for preoperative identification of stages I-II and III-IV of esophageal cancer.

OBJECTIVE: To predict preoperative staging using a radiomics approach based on computed tomography (CT) images of patients with esophageal squamous cell carcinoma (ESCC). METHODS: This retrospective study included 154 patients (primary cohort: n=114; validation cohort: n=40) with pathologically confirmed ESCC. All patients underwent a preoperative CT scan from the neck to abdomen. High throughput and quantitative radiomics features were extracted from the CT images for each patient. A radiomics signature was constructed using the least absolute shrinkage and selection operator (Lasso). Associations between radiomics signature, tumor volume and ESCC staging were explored. Diagnostic performance of radiomics approach and tumor volume for discriminating between stages I-II and III-IV was evaluated and compared using the receiver operating characteristics (ROC) curves and net reclassification improvement (NRI). RESULTS: A total of 9,790 radiomics features were extracted. Ten features were selected to build a radiomics signature after feature dimension reduction. The radiomics signature was significantly associated with ESCC staging (P<0.001), and yielded a better performance for discrimination of early and advanced stage ESCC compared to tumor volume in both the primary [area under the receiver operating characteristic curve (AUC): 0.795vs. 0.694, P=0.003; NRI=0.424)] and validation cohorts (AUC: 0.762 vs. 0.624, P=0.035; NRI=0.834). CONCLUSIONS: The quantitative approach has the potential to identify stage I-II and III-IV ESCC before treatment.

Comparison of microvascular perfusion evaluation among IVIM-DWI, CT perfusion imaging and histological microvessel density in rabbit liver VX2 tumors.

OBJECT: To explore microcirculation features with intravoxel incoherent motion (IVIM) and to compare IVIM with CT perfusion imaging (CTPI) and microvessel density (MVD). MATERIALS AND METHODS: Hepatic CTPI and IVIM were performed in 16 rabbit liver VX2 tumor models. Hepatic arterial perfusion (HAP), hepatic arterial perfusion index (HPI), Blood flow (BF), and blood volume (BV) from CTPI were measured. Apparent diffusion coefficient (ADC), true diffusion coefficient (D), perfusion fraction (f), and pseudo-diffusion coefficient (D*) from IVIM were measured. MVD was counted with CD34 stain. The microcirculation features with IVIM were compared with CTPI parameters and MVD. RESULTS: Strong linear correlations were found between D value (0.89±0.21×10-3mm2/s) and HAP (15.83±6.97ml/min/100mg) (r=0.755, P=0.001) and between f value (12.64±6.66%) and BV (9.74±5.04ml/100mg) (r=0.693, P=0.004). Moderate linear correlations were observed between ADC (1.07±0.32×10-3mm2/s) and HAP (r=0.538, P=0.039), respectively; and between D value and MVD (9.31±2.57 vessels at 400×magnification) (r=0.509, P=0.044). No correlations were found between D* (119.90±37.67×10-3mm2/s) and HAP, HPI (68.34±12.91%), BF (4.95±2.16ml/min/100mg), BV. CONCLUSION: IVIM parameters can characterize microcirculation to certain extent and separate it from pure water molecular diffusion. There is fair correlation between D or ADC value and CTPI parameters or MVD, but no correlation between D* or f value and CTPI parameters or MVD except f value and BV, which is still unclear and need further clinical studies to validate.

Rapid evolution of the Photosystem II electronic structure during water splitting.

Photosynthetic water oxidation is a fundamental process that sustains the biosphere. A Mn4Ca cluster embedded in the photosystem II protein environment is responsible for the production of atmospheric oxygen. Here, time-resolved x-ray emission spectroscopy (XES) was used to observe the process of oxygen formation in real time. These experiments reveal that the oxygen evolution step, initiated by three sequential laser flashes, is accompanied by rapid (within 50 µs) changes to the Mn Kß XES spectrum. However, no oxidation of the Mn4Ca core above the all MnIV state was detected to precede O-O bond formation, and the observed changes were therefore assigned to O-O bond formation dynamics. We propose that O-O bond formation occurs prior to the transfer of the final (4th) electron from the Mn4Ca cluster to the oxidized tyrosine YZ residue. This model resolves the kinetic limitations associated with O-O bond formation, and suggests an evolutionary adaptation to avoid releasing of harmful peroxide species.

Angiomyolipoma with minimal fat: differentiation from clear cell renal cell carcinoma and papillary renal cell carcinoma by texture analysis on CT images.

RATIONALE AND OBJECTIVES: To retrospectively evaluate the diagnostic performance of texture analysis (TA) for the discrimination of angiomyolipoma (AML) with minimal fat, clear cell renal cell cancer (ccRCC), and papillary renal cell cancer (pRCC) on computed tomography (CT) images and to determine the scanning phase, which contains the strongest discriminative power. MATERIALS AND METHODS: Patients with pathologically proved AMLs (n = 18) lacking visible macroscopic fat at CT and patients with pathologically proved ccRCCs (n = 18) and pRCCs (n = 14) were included. All patients underwent CT scan with three phases (precontrast phase [PCP], corticomedullary phase [CMP], and nephrographic phase [NP]). The selected images were analyzed and classified with TA software (MaZda). Texture classification was performed for 1) minimal fat AML versus ccRCC, 2) minimal fat AML versus pRCC, and 3) ccRCC versus pRCC. The classification results were arbitrarily divided into several levels according to the misclassification rates: excellent (misclassification rates ≤10%), good (10%< misclassification rates ≤20%), moderate (20%< misclassification rates ≤30%), fair (30%< misclassification rates ≤40%), and poor (misclassification rates ≥40%). RESULTS: Excellent classification results (error of 0.00%-9.30%) were obtained with nonlinear discriminant analysis for all the three groups, no matter which phase was used. On comparison of the three scanning phases, we observed a trend toward better lesion classification with PCP for minimal fat AML versus ccRCC, CMP, and NP images for ccRCC versus pRCC and found similar discriminative power for minimal fat AML versus pRCC. CONCLUSIONS: TA might be a reliable quantitative method for the discrimination of minimal fat AML, ccRCC, and pRCC.

Oxygen-assisted water partial dissociation on copper: a model study.

It is essential to understand and control the O-H bond cleavage on metal surfaces with pre-adsorbed oxygen atoms in heterogeneous catalytic processes. The adsorption and dissociation of water on clean and oxygen-pre-adsorbed copper surfaces, including Cu(111), Cu(110), Cu(100), Cu(210), Cu(211), Cu(310) and Cu(110)-(1 × 2), as well as Cu-ad-row and Cu-ad-atom, have been investigated by the density functional theory-generalized gradient approximation (DFT-GGA) method. The calculation results indicate that the presence of oxygen species significantly promotes the water dissociation. It is found that the promotion effect depends both on the adsorption energy of the pre-adsorbed oxygen and the distance between the pre-adsorbed oxygen and the stripped hydrogen in water: the more strongly the oxygen atom binds to the metal surface, the less the promotion effect it has on the water O-H bond cleavage; the shorter the distance between pre-adsorbed oxygen and hydrogen in water, the greater is the promotion effect. Based on electronic analysis, physical origin of the promotion effect can be attributed to the strong interaction of acid-base pair sites on oxygen-metal systems.

Diagnostic performance of CT and MRI in distinguishing intraductal papillary neoplasm of the bile duct from cholangiocarcinoma with intraductal papillary growth.

OBJECTIVES: We aimed to evaluate the diagnostic performance of CT and MRI for distinguishing intraductal papillary neoplasm of the bile duct (IPNB) from cholangiocarcinoma (CC) with intraductal papillary growth (IPG). METHODS: Forty-two patients with either IPNB or CC with IPG proven by histopathology were independently reviewed in retrospect. Strict criteria for diagnosis of IPNB included presence of the designated imaging features as follows: local dilatation of the bile duct, nodule within the dilated bile duct, growing along the interior wall of bile duct. Any lesion that was not consistent with the criteria was classified as CC with IPG. Sensitivity, specificity, positive and negative predictive values for characterization of IPNB were calculated, and k test was used to assess the level of agreement. RESULTS: Two imaging reviewers correctly identified 21 of 26 (80.8%) and 22 of 26 (84.6%) IPNB cases, respectively. Alternatively, they correctly identified 14 of 16 (87.5%) and 15 of 16 (93.8%) CC with IPG, respectively. Agreement between the two reviewers was perfect (k = 0.81) for the diagnosis of IPNB and differentiation from CC with IPG. CONCLUSIONS: By using our designated diagnostic criteria of CT and MRI, IPNB can be accurately identified and possible to be distinguished from CC with IPG. KEY POINTS: • IPNB can accurately be identified by using defined diagnostic criteria at CT/MRI. • IPNB has some characteristic CT and MR imaging features. • IPNB is a rare entity; up until now it might have been misdiagnosed.

MRI quantification of non-Gaussian water diffusion in normal human kidney: a diffusional kurtosis imaging study.

Our aim was to prospectively evaluate the feasibility of diffusional kurtosis imaging (DKI) in normal human kidney and to report preliminary DKI measurements. Institutional review board approval and informed consent were obtained. Forty-two healthy volunteers underwent diffusion-weighted imaging (DWI) scans with a 3-T MR scanner. b values of 0, 500 and 1000 s/mm(2) were adopted. Maps of fractional anisotropy (FA), mean diffusivity (MD), radial diffusivity (D⊥), axial diffusivity (D||), mean kurtosis (MK), radial kurtosis (K⊥) and axial kurtosis (K||) were produced. Three representative axial slices in the upper pole, mid-zone and lower pole were selected in the left and right kidney. On each selected slice, three regions of interest were drawn on the renal cortex and another three on the medulla. Statistical comparison was performed with t-test and analysis of variance. Thirty-seven volunteers successfully completed the scans. No statistically significant differences were observed between the left and right kidney for all metrics (p values in the cortex: FA, 0.114; MD, 0.531; D⊥, 0.576; D||, 0.691; MK, 0.934; K⊥, 0.722; K||, 0.891; p values in the medulla: FA, 0.348; MD, 0.732; D⊥, 0.470; D||, 0.289; MK, 0.959; K⊥, 0.780; K||, 0.287). Kurtosis metrics (MK, K||, K⊥) obtained in the renal medulla were significantly (p <0.001) higher than those in the cortex (0.552 ± 0.04, 0.637 ± 0.07 and 0.530 ± 0.08 in the medulla and 0.373 ± 0.04, 0.492 ± 0.06 and 0.295 ± 0.06 in the cortex, respectively). For the diffusivity measures, FA of the medulla (0.356 ± 0.03) was higher than that of the cortex (0.179 ± 0.03), whereas MD, D⊥ and D|| (mm(2) /ms) were lower in the medulla than in the cortex (3.88 ± 0.09, 3.50 ± 0.23 and 4.65 ± 0.29 in the cortex and 2.88 ± 0.11, 2.32 ± 0.20 and 3.47 ± 0.31 in the medulla, respectively). Our results indicate that DKI is feasible in the human kidney. We have reported the preliminary DKI measurements of normal human kidney that demonstrate well the non-Gaussian behavior of water diffusion, especially in the renal medulla.

Structural characterization of CO-inhibited Mo-nitrogenase by combined application of nuclear resonance vibrational spectroscopy, extended X-ray absorption fine structure, and density functional theory: new insights into the effects of CO binding and the role of the interstitial atom.

The properties of CO-inhibited Azotobacter vinelandii (Av) Mo-nitrogenase (N2ase) have been examined by the combined application of nuclear resonance vibrational spectroscopy (NRVS), extended X-ray absorption fine structure (EXAFS), and density functional theory (DFT). Dramatic changes in the NRVS are seen under high-CO conditions, especially in a 188 cm(-1) mode associated with symmetric breathing of the central cage of the FeMo-cofactor. Similar changes are reproduced with the α-H195Q N2ase variant. In the frequency region above 450 cm(-1), additional features are seen that are assigned to Fe-CO bending and stretching modes (confirmed by (13)CO isotope shifts). The EXAFS for wild-type N2ase shows evidence for a significant cluster distortion under high-CO conditions, most dramatically in the splitting of the interaction between Mo and the shell of Fe atoms originally at 5.08 Å in the resting enzyme. A DFT model with both a terminal -CO and a partially reduced -CHO ligand bound to adjacent Fe sites is consistent with both earlier FT-IR experiments, and the present EXAFS and NRVS observations for the wild-type enzyme. Another DFT model with two terminal CO ligands on the adjacent Fe atoms yields Fe-CO bands consistent with the α-H195Q variant NRVS. The calculations also shed light on the vibrational "shake" modes of the interstitial atom inside the central cage, and their interaction with the Fe-CO modes. Implications for the CO and N2 reactivity of N2ase are discussed.

Spectroscopic analysis of catalytic water oxidation by [Ru(II)(bpy)(tpy)H2O]2+ suggests that Ru(V)═O is not a rate-limiting intermediate.

Modern chemistry's grand challenge is to significantly improve catalysts for water splitting. Further progress requires detailed spectroscopic and computational characterization of catalytic mechanisms. We analyzed one of the most studied homogeneous single-site Ru catalysts, [Ru(II)(bpy)(tpy)H2O](2+) (where bpy = 2,2'-bipyridine, tpy = 2,2';6',2″-terpyridine). Our results reveal that the [Ru(V)(bpy)(tpy)═O](3+) intermediate, reportedly detected in catalytic mixtures as a rate-limiting intermediate in water activation, is not present as such. Using a combination of electron paramagnetic resonance (EPR) and X-ray absorption spectroscopy, we demonstrate that 95% of the Ru complex in the catalytic steady state is of the form [Ru(IV)(bpy)(tpy)═O](2+). [Ru(V)(bpy)(tpy)═O](3+) was not observed, and according to density functional theory (DFT) analysis, it might be thermodynamically inaccessible at our experimental conditions. A reaction product with unique EPR spectrum was detected in reaction mixtures at about 5% and assigned to Ru(III)-peroxo species with (-OOH or -OO- ligands). We also analyzed the [Ru(II)(bpy)(tpy)Cl](+) catalyst precursor and confirmed that this molecule is not a catalyst and its oxidation past Ru(III) state is impeded by a lack of proton-coupled electron transfer. Ru-Cl exchange with water is required to form active catalysts with the Ru-H2O fragment. [Ru(II)(bpy)(tpy)H2O](2+) is the simplest representative of a larger class of water oxidation catalysts with neutral, nitrogen containing heterocycles. We expect this class of catalysts to work mechanistically in a similar fashion via [Ru(IV)(bpy)(tpy)═O](2+) intermediate unless more electronegative (oxygen containing) ligands are introduced in the Ru coordination sphere, allowing the formation of more oxidized Ru(V) intermediate.

Serial time-resolved crystallography of photosystem II using a femtosecond X-ray laser.

Photosynthesis, a process catalysed by plants, algae and cyanobacteria converts sunlight to energy thus sustaining all higher life on Earth. Two large membrane protein complexes, photosystem I and II (PSI and PSII), act in series to catalyse the light-driven reactions in photosynthesis. PSII catalyses the light-driven water splitting process, which maintains the Earth's oxygenic atmosphere. In this process, the oxygen-evolving complex (OEC) of PSII cycles through five states, S0 to S4, in which four electrons are sequentially extracted from the OEC in four light-driven charge-separation events. Here we describe time resolved experiments on PSII nano/microcrystals from Thermosynechococcus elongatus performed with the recently developed technique of serial femtosecond crystallography. Structures have been determined from PSII in the dark S1 state and after double laser excitation (putative S3 state) at 5 and 5.5 Å resolution, respectively. The results provide evidence that PSII undergoes significant conformational changes at the electron acceptor side and at the Mn4CaO5 core of the OEC. These include an elongation of the metal cluster, accompanied by changes in the protein environment, which could allow for binding of the second substrate water molecule between the more distant protruding Mn (referred to as the 'dangler' Mn) and the Mn3CaOx cubane in the S2 to S3 transition, as predicted by spectroscopic and computational studies. This work shows the great potential for time-resolved serial femtosecond crystallography for investigation of catalytic processes in biomolecules.

Liver diffusion-weighted MR imaging: reproducibility comparison of ADC measurements obtained with multiple breath-hold, free-breathing, respiratory-triggered, and navigator-triggered techniques.

PURPOSE: To prospectively compare the reproducibility of normal liver apparent diffusion coefficient (ADC) measurements by using different respiratory motion compensation techniques with multiple breath-hold (MBH), free-breathing (FB), respiratory-triggered (RT), and navigator-triggered (NT) diffusion-weighted (DW) imaging and to compare the ADCs at different liver anatomic locations. MATERIALS AND METHODS: The study protocol was approved by the institutional review board, and written informed consent was obtained from each participant. Thirty-nine volunteers underwent liver DW imaging twice. Imaging was performed with a 1.5-T MR imager with MBH, FB, RT, and NT techniques (b = 0, 100, and 500 sec/mm(2)). Three representative sections--superior, central, and inferior--were selected on left and right liver lobes, respectively. On each selected section, three regions of interest were drawn, and ADCs were measured. Analysis of variance was used to assess ADCs among the four techniques and various anatomic locations. Reproducibility of ADCs was assessed with the Bland-Altman method. RESULTS: ADCs obtained with MBH (range: right lobe, [1.641-1.662] × 10(-3)mm(2)/sec; left lobe, [2.034-2.054] ×10(-3)mm(2)/sec) were higher than those obtained with FB (right, [1.349-1.391] ×10(-3)mm(2)/sec; left, [1.630-1.700] ×10(-3)mm(2)/sec), RT (right, [1.439-1.455] ×10(-3)mm(2)/sec; left, [1.720-1.755] ×10(-3)mm(2)/sec), or NT (right, [1.387-1.400] ×10(-3)mm(2)/sec; left, [1.661-1.736] ×10(-3)mm(2)/sec) techniques (P < .001); however, no significant difference was observed between ADCs obtained with FB, RT, and NT techniques (P = .130 to P >.99). ADCs showed a trend to decrease moving from left to right. Reproducibility in the left liver lobe was inferior to that in the right, and the central middle segment in the right lobe had the most reproducible ADC. Statistical differences in ADCs were observed in the left-right direction in the right lobe (P < .001), but they were not observed in the superior-inferior direction (P = .144-.450). However, in the left liver lobe, statistical differences existed in both directions (P = .001 to P = .016 in the left-right direction, P < .001 in the superior-inferior direction). CONCLUSION: Both anatomic location and DW imaging technique influence liver ADC measurements and their reproducibility. FB DW imaging is recommended for liver DW imaging because of its good reproducibility and shorter acquisition time compared with that of MBH, RT, and NT techniques.

[Preparation of new solid-phase microextraction fiber based on cucurbit[7] uril coordination polymers and its application in determination of polycyclic aromatic hydrocarbons from environmental water samples].

A novel solid-phase microextraction fiber was prepared via simple adhesion process. Fused-silica fiber was used as supporting substrate and the cucurbit[7] uril/Eu coordination polymers induced by [CdCl4]2- anion was affixed onto the surface of fused-silica fiber with high-temperature epoxy resin as a sorbent coating. The SPME coating was characterized by scanning electron microscopy and thermal analysis. Coupled to gas chromatography, the fiber was used to extract polycyclic aromatic hydrocarbons (PAHs) in aqueous samples. Analytical performances of the proposed method were investigated under the optimum extraction conditions (extraction temperature, 75 °C; mass concentration of NaCl, 200 g/L; extraction time, 40 min; desorption time, 2 min; desorption temperature, 250 °C) compared with commercial fibers for the same analytes. For the seven PAHs (naphthalene, acenaphthene, fluorene, phenanthrene, anthracene, fluoranthene and pyrene), good linearity was obtained ranging from 1 µg/L to 1000 µg/L. Limits of detection were 0.29-2.09 µg/L with the RSDs no more than 8.6%. The established SPME-GC method was applied to determine the spiked PAHs in real river water samples and satisfactory results were obtained with recoveries between 97. 2% and 109. 0%. Under the optimal conditions, the extraction effect of home-made coating was similar to the commercial PDMS (polydimethylsiloxane) coating, which further confirmed its potential in the solid-phase microextraction.

IR-monitored photolysis of CO-inhibited nitrogenase: a major EPR-silent species with coupled terminal CO ligands.

Fourier transform infrared spectroscopy (FTIR) was used to observe the photolysis and recombination of a new EPR-silent CO-inhibited form of α-H195Q nitrogenase from Azotobacter vinelandii. Photolysis at 4 K reveals a strong negative IR difference band at nu = 1938 cm(-1), along with a weaker negative feature at 1911 cm(-1). These bands and the associated chemical species have both been assigned the label "Hi-3". A positive band at nu = 1921 cm(-1) was assigned to the "Lo-3" photoproduct. By using an isotopic mixture of (12)C (16)O and (13)C (18)O, we show that the Hi-3 bands arise from coupling of two similar CO oscillators with one uncoupled frequency at approximately nu = 1917 cm(-1). Although in previous studies Lo-3 was not observed to recombine, by extending the observation range to 200-240 K, we found that recombination to Hi-3 does indeed occur, with an activation energy of approximately 6.5 kJ mol(-1). The frequencies of the Hi-3 bands suggest terminal CO ligation. This hypothesis was tested with DFT calculations on models with terminal CO ligands on Fe2 and Fe6 of the FeMo-cofactor. An S = 0 model with both CO ligands in exo positions predicts symmetric and asymmetric stretches at nu = 1938 and 1909 cm(-1), respectively, with relative band intensities of about 3.5:1, which is in good agreement with experiment. From the observed IR intensities, Hi-3 was found to be present at a concentration about equal to that of the EPR-active Hi-1 species. The relevance of Hi-3 to the nitrogenase catalytic mechanism and its recently discovered Fischer-Tropsch chemistry is discussed.