Development of Inhibitory Compounds for Metallo-beta-lactamase through Computational Design and Crystallographic Analysis.

Metallo-ß-lactamases (MBL) deactivate ß-lactam antibiotics through a catalytic reaction caused by two zinc ions at the active center. Since MBLs deteriorate a wide range of antibiotics, they are dangerous factors for bacterial multidrug resistance. In this work, organic synthesis, computational design, and crystal structure analysis were performed to obtain potent MBL inhibitors based on a previously identified hit compound. The hit compound comprised 3,4-dihydro-2(1H)-quinolinone linked with a phenyl-ether-methyl group via a thiazole ring. In the first step, the thiazole ring was replaced with a tertiary amine to avoid the planar structure. In the second step, we virtually modified the compound by keeping the quinolinone backbone. Every modified compound was bound to a kind of MBL, imipenemase-1 (IMP-1), and the binding pose was optimized by a molecular mechanics calculation. The binding scores were evaluated for the respective optimized binding poses. Given the predicted binding poses and calculated binding scores, candidate compounds were determined for organic syntheses. The inhibitory activities of the synthesized compounds were measured by an in vitro assay for two kinds of MBLs, IMP-1 and New Delhi metallo-ß-lactamase (NDM-1). A quinolinone connected with an amine bound with methyl-phenyl-ether-propyl and cyclohexyl-ethyl showed a 50% inhibitory concentration of 4.8 µM. An X-ray crystal analysis clarified the binding structure of a synthesized compound to IMP-1. The δ-lactam ring of quinolinone was hydrolyzed, and the generated carboxyl group was coordinated with zinc ions. The findings on the chemical structure and binding pose are expected to be a base for developing MBL inhibitors.

Perioperative proximal splenic artery embolization in cirrhotic patients with splenomegaly.

INTRODUCTION: The purpose of this study was to determine the effect of proximal splenic artery embolization (SAE) in cirrhotic patients with splenomegaly who underwent surgical laparotomy. MATERIAL AND METHODS: This retrospective observational study included 8 cirrhotic patients with splenomegaly. They underwent proximal SAE before- (n = 6) or after (n = 2) laparotomy. Vascular plugs or coils were placed in the proximal splenic artery. The diameter of the portal vein and the splenic volume were recorded. Clinical outcome assessments included platelet counts, the model for end-stage liver disease (MELD) score, and complications. RESULTS: After embolization, the portal venous diameter was significantly smaller (pre: 13.6 ± 2.7 mm, post: 12.5 ± 2.3 mm, p = 0.023), the splenic volume was significantly decreased (pre: 463.2 ± 145.7 ml, post: 373.3 ± 108.5 ml, p = 0.008) and the platelet count was significantly higher (pre: 69.6 ± 30.8 × 103/µl, post: 86.8 ± 27.7 × 103/µl, p = 0.035). Before embolization, the median MELD score was 12; after embolization, it was 11 (p = 0.026). No patient developed post-treatment complications after embolization. CONCLUSIONS: The reduction of hypersplenism by perioperative proximal SAE may be safe and reduce the surgical risk in cirrhotic patients with splenomegaly.

Iodine maps derived from sparse-view kV-switching dual-energy CT equipped with a deep learning reconstruction for diagnosis of hepatocellular carcinoma.

Deep learning-based spectral CT imaging (DL-SCTI) is a novel type of fast kilovolt-switching dual-energy CT equipped with a cascaded deep-learning reconstruction which completes the views missing in the sinogram space and improves the image quality in the image space because it uses deep convolutional neural networks trained on fully sampled dual-energy data acquired via dual kV rotations. We investigated the clinical utility of iodine maps generated from DL-SCTI scans for assessing hepatocellular carcinoma (HCC). In the clinical study, dynamic DL-SCTI scans (tube voltage 135 and 80 kV) were acquired in 52 patients with hypervascular HCCs whose vascularity was confirmed by CT during hepatic arteriography. Virtual monochromatic 70 keV images served as the reference images. Iodine maps were reconstructed using three-material decomposition (fat, healthy liver tissue, iodine). A radiologist calculated the contrast-to-noise ratio (CNR) during the hepatic arterial phase (CNRa) and the equilibrium phase (CNRe). In the phantom study, DL-SCTI scans (tube voltage 135 and 80 kV) were acquired to assess the accuracy of iodine maps; the iodine concentration was known. The CNRa was significantly higher on the iodine maps than on 70 keV images (p < 0.01). The CNRe was significantly higher on 70 keV images than on iodine maps (p < 0.01). The estimated iodine concentration derived from DL-SCTI scans in the phantom study was highly correlated with the known iodine concentration. It was underestimated in small-diameter modules and in large-diameter modules with an iodine concentration of less than 2.0 mgI/ml. Iodine maps generated from DL-SCTI scans can improve the CNR for HCCs during hepatic arterial phase but not during equilibrium phase in comparison with virtual monochromatic 70 keV images. Also, when the lesion is small or the iodine concentration is low, iodine quantification may result in underestimation.

Utility of Wavelet Denoising with Geometry Factor Weighting for Gadoxetic Acid-enhanced Hepatobiliary-phase MR Imaging.

PURPOSE: The wavelet denoising with geometry factor weighting (g-denoising) method can reduce the image noise by adapting to spatially varying noise levels induced by parallel imaging. The aim of this study was to investigate the clinical applicability of g-denoising on hepatobiliary-phase (HBP) images with gadoxetic acid. METHODS: We subjected 53 patients suspected of harboring hepatic neoplastic lesions to gadoxetic acid-enhanced HBP imaging with and without g-denoising (g+HBP and g-HBP). The matrix size was reduced for g+HBP images to avoid prolonging the scanning time. Two radiologists calculated the SNR, the portal vein-, and paraspinal muscle contrast-to-noise ratio (CNR) relative to the hepatic parenchyma (liver-to-portal vein- and liver-to-muscle CNR). Two other radiologists independently graded the sharpness of the liver edge, the visibility of intrahepatic vessels, the image noise, the homogeneity of liver parenchyma, and the overall image quality using a 5-point scale. Differences between g-HBP and g+HBP images were determined with the two-sided Wilcoxon signed-rank test. RESULTS: The liver-to-portal- and liver-to-muscle CNR and the SNR were significantly higher on g+HBP- than g-HBP images (P < 0.01), as was the qualitative score for the image noise, homogeneity of liver parenchyma, and overall image quality (P < 0.01). Although there were no significant differences in the scores for the sharpness of the liver edge or the score assigned for the visibility of intrahepatic vessels (P = 0.05, 0.43), with g+HBP the score was lower in three patients for the sharpness of the liver edge and in six patients for the visibility of intrahepatic vessels. CONCLUSION: At gadoxetic acid-enhanced HBP imaging, g-denoising yielded a better image quality than conventional HBP imaging although the anatomic details may be degraded.

An introduction to photon-counting detector CT (PCD CT) for radiologists.

The basic performance of photon-counting detector computed tomography (PCD CT) is superior to conventional CT (energy-integrating detector CT: EID CT) because its spatial- and contrast resolution of soft tissues is higher, and artifacts are reduced. Because the X-ray photon energy separation is better with PCD CT than conventional EID-based dual-energy CT, it has the potential to improve virtual monochromatic- and virtual non-contrast images, material decomposition including quantification of the iodine distribution, and K-edge imaging. Therefore, its clinical applicability may be increased. Although the image quality of PCD CT scans is superior to that of EID CT currently, further improvement may be possible. The introduction of iterative image reconstruction and reconstruction with deep convolutional neural networks will be useful.

Understanding CT imaging findings based on the underlying pathophysiology in patients with small bowel ischemia.

Because acute small bowel ischemia has a high mortality rate, it requires rapid intervention to avoid unfavorable outcomes. Computed tomography (CT) examination is important for the diagnosis of bowel ischemia. Acute small bowel ischemia can be the result of small bowel obstruction or mesenteric ischemia, including mesenteric arterial occlusion, mesenteric venous thrombosis, and non-occlusive mesenteric ischemia. The clinical significance of each CT finding is unique and depends on the underlying pathophysiology. This review describes the definition and mechanism(s) of bowel ischemia, reviews CT findings suggesting bowel ischemia, details factors involved in the development of small bowel ischemia, and presents CT findings with respect to the different factors based on the underlying pathophysiology. Such knowledge is needed for accurate treatment decisions.

Bioleaching of tennantite concentrate: influence of microbial community and solution redox potential.

Despite its growing importance as a Cu resource, studies on tennantite bioleaching are highly limited. One of the key challenges in processing such Cu-As sulfides is their refractoriness and the solubilisation of toxic As. The ultimate goal is to achieve selective bioleaching of Cu with simultaneous immobilisation of As in the leach residues. This study investigated the effectiveness of activated carbon (AC)-assisted bioleaching of tennantite concentrate using a mixed culture containing various "strong" and "weak" Fe-oxidising bacteria/archaea plus a S-oxidising bacterium, with particular emphasis on controlling the solution redox potential (Eh). In the initial flask bioleaching tests, a steady increase in Eh (up to 840 mV) was observed, reflecting the activity of "strong" Fe-oxidisers. In this situation, AC dosing effectively suppressed the Eh value and the highest Cu dissolution (70%) was obtained in the AC-0.01% system, while simultaneously immobilising As. In order to maximise Cu dissolution and As immobilisation, it was found preferable to target the Eh range of 650-700 mV during bioleaching. The next bioreactor tests used the mixed culture of the same origin, but had been subcultured a few generations further on tennantite concentrate. The Eh level remained unexpectedly low (~630 mV) for most of the leaching period, regardless of the AC dosage. It was later found that the bioreactor systems were almost exclusively dominated by Sb. thermosulfidooxidans, a "weak" Fe oxidiser with high Cu/As tolerance. In this case, there was no need to artificially suppress the Eh level by AC dosing and Cu leached readily to a final Cu dissolution of ~60% while As dissolution was suppressed to ~15%. Thus, depending on the microbial community that develops at the processing site, Eh control can be achieved either naturally by the activity of "weak" Fe-oxidisers as the predominant survivors under high Cu/As stress, or artificially by the addition of an Eh regulator such as a carbon catalyst.

Global illumination rendering versus volume rendering for the forensic evaluation of stab wounds using computed tomography.

We compared three-dimensional (3D) CT images of stabbing victims subjected to volume-rendering (VR) or global illumination-rendering (GIR), a new technique now available for the reconstruction of 3D CT images. It simulates the complete interactions of photons with the scanned object, thereby providing photorealistic images. The diagnostic value of the images was also compared with that of macroscopic photographs. We used postmortem 3D CT images of 14 stabbing victims who had undergone autopsy and CT studies. The 3D CT images were subjected to GIR or VR and the 3D effect and the smoothness of the skin surface were graded on a 5-point scale. We also compared the 3D CT images of 37 stab wounds with macroscopic photographs. The maximum diameter of the wounds was measured on VR and GIR images and compared with the diameter recorded at autopsy. The overall image-quality scores and the ability to assess the stab wounds were significantly better on GIR than VR images (median scores: VR = 3 vs GIR = 4, p < 0.01). The mean difference between the wound diameter measured on VR and GIR images and at autopsy were both 0.2 cm, respectively. For the assessment of stab wounds, 3D CT images subjected to GIR were superior to VR images. The diagnostic value of 3D CT GIR image was comparable to that of macroscopic photographs.

Identification of the Inhibitory Compounds for Metallo-ß-lactamases and Structural Analysis of the Binding Modes.

Metallo-ß-lactamases (MBLs) are significant threats to humans because they deteriorate many kinds of ß-lactam antibiotics and are key enzymes responsible for multi-drug resistance of bacterial pathogens. As a result of in vitro screening, two compounds were identified as potent inhibitors of two kinds of MBLs: imipenemase (IMP-1) and New Delhi metallo-ß-lactamase (NDM-1). The binding structure of one of the identified compounds was clarified by an X-ray crystal analysis in complex with IMP-1, in which two possible binding poses were observed. Molecular dynamics (MD) simulations were performed by building two calculation models from the respective binding poses. The compound was stably bound to the catalytic site during the simulation in one pose. The binding model between NDM-1 and the compound was constructed for MD simulation. Calculation results for NDM-1 were similar to those of IMP-1. The simulation suggested that the binding of the identified inhibitory compound was also durable in the catalytic site of NDM-1. The compound will be a sound basis for the development of the inhibitors for MBLs.

Advanced CT techniques for assessing hepatocellular carcinoma.

Hepatocellular carcinoma (HCC) is the sixth-most common cancer in the world, and hepatic dynamic CT studies are routinely performed for its evaluation. Ongoing studies are examining advanced imaging techniques that may yield better findings than are obtained with conventional hepatic dynamic CT scanning. Dual-energy CT-, perfusion CT-, and artificial intelligence-based methods can be used for the precise characterization of liver tumors, the quantification of treatment responses, and for predicting the overall survival rate of patients. In this review, the advantages and disadvantages of conventional hepatic dynamic CT imaging are reviewed and the general principles of dual-energy- and perfusion CT, and the clinical applications and limitations of these technologies are discussed with respect to HCC. Finally, we address the utility of artificial intelligence-based methods for diagnosing HCC.

Does chemical shift imaging offer a biomarker for the diagnosis and assessment of disease severity in multiple myeloma?: A cross-sectional study.

ABSTRACT: To investigate whether chemical shift imaging (CSI) is useful for differentiating myelomatous infiltration from hematopoietic bone marrow (BM) and for quantitatively assessing disease severity.In this retrospective study, spinal MRI, including a sagittal iterative decomposition of water and fat with echo asymmetry and least-squares estimation T2 fast spin-echo sequence, was performed on 76 myeloma patients (45 men, 67.0 ±â€Š11.4 years; 31 women, 66.5 ±â€Š11.0 years) and 30 control subjects (20 men, 67.0 ±â€Š8.4 years; 10 women, 67.0 ±â€Š9.2 years). The fat-signal fraction (FF) and mean signal dropout ratio (DR) were calculated from lumbar BM that contained no focal lesions. The BM plasma cell percentage (BMPC%) and serological data were obtained. As DR is highest when FF = 50%, the patients were divided into 2 groups: a water-dominant group (FF < 50%) and a fat-dominant group (FF > 50%).Serum monoclonal protein (M protein), ß2-microglobulin, and BMPC% were significantly higher in the water-dominant group than in the fat-dominant group. In the water-dominant group, DR correlated significantly with BMPC% and M protein, whereas in the control group, DR showed a weak correlation with age but no correlation with other clinical factors. No significant differences in any clinical data were seen between high and low DR.CSI proved ineffective for differentiating myelomatous infiltration from hematopoietic BM. For myeloma patients with relatively high BM cellularity, a small signal drop on opposed-phase images indicated a higher tumor burden. For BM with relatively low cellularity, disease severity was not reflected by CSI.

Assessment of early treatment response on MRI in multiple myeloma: Comparative study of whole-body diffusion-weighted and lumbar spinal MRI.

OBJECTIVES: To compare remission status at completion of chemotherapy for multiple myeloma (MM) with changes in total diffusion volume (tDV) calculated from whole-body diffusion-weighted imaging (WB-DWI) and fat fraction (FF) of lumbar bone marrow (BM) by modified Dixon Quant (mDixon Quant) soon after induction of chemotherapy, and to assess the predictive value of MRI. METHODS: Fifty patients (mean age, 66.9 ± 10.5 years) with symptomatic myeloma were examined before and after two cycles of chemotherapy. From WB-DWI data, tDV was obtained with the threshold for positive BM involvement. Mean FF was calculated from lumbar BM using the mDixon Quant sequence. At the completion of chemotherapy, patients were categorized into a CR/very good PR (VGPR) group (n = 15; mean age, 67.6 ± 10.3 years) and a PR, SD or PD group (n = 35; mean age, 69.1 ± 8.6 years). ROC curves were plotted to assess performance in predicting achievement of CR/VGPR. RESULTS: At second examination, serum M protein, ß2-microglobulin, and tDV were significantly decreased and hemoglobin, mean ADC, and FF were significantly increased in the CR/VGPR group and serum M protein was significantly increased in the PR/SD/PD group. The general linear model demonstrated that percentage changes in FF and M protein contributed significantly to achieving CR/VGPR (P = 0.02, P = 0.04, respectively). AUCs of ROC curves were 0.964 for FF and 0.847 for M protein. CONCLUSIONS: Early change in FF of lumbar BM and serum M protein soon after induction of chemotherapy contributed significantly to prediction of CR/VGPR.

Tris-(4,4'-di-tert-butyl-2,2'-bi-pyridine)(trans-4-tert-butyl-cyclo-hexa-nolato)-deca-µ-oxido-hepta-oxido-hepta-vanadium aceto-nitrile monosolvate including another unknown solvent mol-ecule.

The title hepta-nuclear alkoxido(oxido)vanadium(V) oxide cluster complex, [V7(C10H19O)O17(C18H24N2)3]·CH3CN, was obtained by the reaction of [V8O20(C18H24N2)4] with 4-tert-butyl-cyclo-hexa-nol (mixture of cis and trans) in a mixed CHCl3/CH3CN solvent. The complex has a V7O18N6 core with approximately C s symmetry, which is composed of two VO4 tetra-hedra, two VO6 octa-hedra and three VO4N2 octa-hedra. In the crystal, these complexes are linked together by weak inter-molecular C-H⋯O hydrogen bonds between the 4,4'-di-tert-butyl-2,2'-bi-pyridine ligand and the V7O18N6 core, forming a one-dimensional network along the c-axis direction. Besides the complex, the asymmetric unit contains one CH3CN solvent mol-ecule. The contribution of other disordered solvent mol-ecules to the scattering was removed using the SQUEEZE option in PLATON [Spek (2015 ▸). Acta Cryst. C71, 9-18]. The unknown solvent mol-ecules are not considered in the chemical formula and other crystal data.

Biochemical analysis of GTPase FlhF which controls the number and position of flagellar formation in marine Vibrio.

FlhF controls the number and position of the polar flagellar formation of Vibrio species. FlhF, is a paralog of FtsY, a GTPase acting in the Sec membrane transport system of bacteria, and localizes at the cell pole. Mutations in the conserved GTPase motif of FlhF lost polar localization capability and flagellar formation. Vibrio FlhF has not, until now, been purified as soluble protein. Here, we report that addition of MgCl2 and GTP or GDP at the step of cell lysis greatly improved the solubility of FlhF, allowing us to purify it in homogeneity. Purified FlhF showed GTPase activity only in the presence of FlhG. Of twelve FlhF GTPase motif mutants showing reduced function, eleven were recovered as precipitate after the cell disruption. The E440K substitution could be purified and showed no GTPase activity even in the presence of FlhG. Interestingly an FlhF substitution in the putative catalytic residue for GTP hydrolysis, R334A, allowed normal flagellar formation although GTPase activity of FlhF was completely abolished. Furthermore, size exclusion chromatography of purified FlhF revealed that it forms dimers in the presence of GTP but exists as monomer in the presence of GDP. We speculate that the GTP binding allows FlhF to dimerize and localize at the pole where it initiates flagellar formation, and the GDP-bound form diffuses as monomer.

Feasibility and Safety of CT-guided Intrathoracic and Bone Re-biopsy for Non-small Cell Lung Cancer.

AIM: This study aimed to retrospectively determine the feasibility and safety of computed tomography (CT)-guided intrathoracic and bone re-biopsy for patients with non-small cell lung cancer (NSCLC). MATERIALS AND METHODS: Seventeen patients underwent CT-guided intrathoracic or bone re-biopsy for the determination of epidermal growth factor receptor (EGFR) T790M mutation and/or programmed cell death-ligand 1 (PD-L1) expression. The characteristics of each lesion, success rate of analyses, and complications were investigated. RESULTS: Specimens from 16 out of the 17 patients were adequate for evaluation of EGFR T790M mutation and/or PD-L1 expression. The mean diameter of the lesions was 40 mm, the mean procedural time was 24 minutes, and the median number of punctures was 2. There were no significant differences in lesion characteristics and success rates between CT-guided intrathoracic and bone re-biopsies. No serious complications occurred. CONCLUSION: Both CT-guided intrathoracic and bone re-biopsies for patients with NSCLC were feasible and safe.

Analysis of the GTPase motif of FlhF in the control of the number and location of polar flagella in Vibrio alginolyticus.

Vibrio alginolyticus normally has a single polar flagellum whose number and placement are regulated positively by FlhF. FlhF is a GTPase and homolog of a signal recognition particle (SRP) protein called Ffh and SRP receptor FtsY. FlhF is located at the cell pole and directs formation of the flagellum. To study the mechanism of FlhF localization, we introduced random mutations into flhF by means of hydroxylamine and isolated mutants that could not generate the flagellum at the cell pole. The novel mutations were only mapped to the GTPase motif of FlhF. The mutant FlhF proteins showed reduced polar localization as compared to the wild type and still could associate with the membrane. These results support the assumption that the GTPase motif of FlhF plays a critical role in the polar localization of this protein during formation of the flagellum.