Biosynthesis and Genome Mining Potentials of Nucleoside Natural Products.

Nucleoside natural products show diverse biological activities and serve as leads for various application purposes, including human and veterinary medicine and agriculture. Studies in the past decade revealed that these nucleosides are biosynthesized through divergent mechanisms, in which early steps of the pathways can be classified into two types (C5' oxidation and C5' radical extension), while the structural diversity is created by downstream tailoring enzymes. Based on this biosynthetic logic, we investigated the genome mining discovery potentials of these nucleosides using the two enzymes representing the two types of C5' modifications: LipL-type α-ketoglutarate (α-KG) and Fe-dependent oxygenases and NikJ-type radical S-adenosyl-L-methionine (SAM) enzymes. The results suggest that this approach allows discovery of putative nucleoside biosynthetic gene clusters (BGCs) and the prediction of the core nucleoside structures. The results also revealed the distribution of these pathways in nature and implied the possibility of future genome mining discovery of novel nucleoside natural products.

Addendum: Cryptic phosphorylation in nucleoside natural product biosynthesis.

A Correction to this paper has been published: https://doi.org/10.1038/s41589-021-00867-7.

Cryptic phosphorylation in nucleoside natural product biosynthesis.

Kinases are annotated in many nucleoside biosynthetic gene clusters but generally are considered responsible only for self-resistance. Here, we report an unexpected 2'-phosphorylation of nucleoside biosynthetic intermediates in the nikkomycin and polyoxin pathways. This phosphorylation is a unique cryptic modification as it is introduced in the third of seven steps during aminohexuronic acid (AHA) nucleoside biosynthesis, retained throughout the pathway's duration, and is removed in the last step of the pathway. Bioinformatic analysis of reported nucleoside biosynthetic gene clusters indicates the presence of cryptic phosphorylation in other pathways and the importance of functional characterization of kinases in nucleoside biosynthetic pathways in general. This study also functionally characterized all of the enzymes responsible for AHA biosynthesis and revealed that AHA is constructed via a unique oxidative C-C bond cleavage reaction. The results indicate a divergent biosynthetic mechanism for three classes of antifungal nucleoside natural products.

Correlation between pretreatment and follow-up infarct volume using CT perfusion imaging: the Bayesian versus singular value decomposition method.

PURPOSE: Pretreatment ischemic core volume is conceptually equal to follow-up infarct volume (FIV) in patients with successful recanalization. However, there is sometimes an absolute volume difference (AD) between pretreatment core volume and FIV. The aim was to compare the AD values between the Bayesian and the singular value decomposition (SVD) methods with time from onset-to-imaging in acute ischemic stroke (AIS) patients undergoing mechanical thrombectomy. METHODS: Consecutive AIS patients were included if they had the following: (1) anterior large vessel occlusion (internal carotid or middle cerebral artery); (2) within 24 h of onset; (3) pretreatment CT perfusion (CTP); (4) successful recanalization (mTICI ≥ 2b); and (5) 24-h diffusion-weighted imaging (DWI). FIV was measured on 24-h DWI. The AD value between FIV and the pretreatment core volume was calculated for Bayesian and SVD methods. Spearman's rank correlation coefficient (rho) was calculated as appropriate. RESULTS: In the 47 patients enrolled (25 men; median age 78 years; median baseline National Institutes of Health Stroke Scale, 22), the median time from onset-to-imaging and onset-to-recanalization was 136 and 220 min, respectively. Shorter onset-to-imaging time was correlated with a larger AD value, and more trend was seen in the SVD method (rho = - 0.28, p = 0.05) compared with the Bayesian method (rho = - 0.08). A larger pretreatment core volume was correlated with a larger AD value, and this tendency was slightly stronger for the SVD (rho = 0.63, p < 0.01) than for the Bayesian (rho = 0.32, p = 0.03) method. CONCLUSIONS: The Bayesian method might be more correlated with FIV than the SVD method in patients with a large ischemic lesion immediately after stroke onset, but not perfect.

Using the pre-freezing technique results in a wider area of antral isolation during cryoballoon pulmonary vein isolation in patients with atrial fibrillation.

The "pre-freezing" technique was a method in which a fully inflated balloon after the start of freezing was pressed against the pulmonary vein (PV) during cryoballoon ablation and has been applied especially in large-size PVs. Of 556 patients who underwent cryoballoon ablation for atrial fibrillation (AF), the pre-freezing technique was applied to 48 patients. The resulting 2:1 propensity score-matched data set included 120 patients. Using the pre-freezing technique, all left-superior PVs, all left-inferior PVs, and 95% of right-superior PVs were successfully isolated. In most right-inferior PVs, complete sealing using the pre-freezing technique was challenging, and this technique was not applied. Procedure time was similar between the two groups. In the pre-freezing group, the percentage of the left atrial posterior wall isolated was larger (47.6 ± 10.3 vs. 42.8 ± 15.7%, P = 0.006), and the postoperative reduction of diaphragmatic compound motor action potentials tended to occur less frequently (2.5 vs. 12.5%, P = 0.07), and the reduction of the cross-sectional LSPV area was smaller (17.5 ± 12.2 vs. 27.2 ± 19.8%, P = 0.03) than the conventional group. The AF-free rate of the two groups was similar between the two groups (P = 0.15). The pre-freezing technique was a simple method that can isolate a wider surface area during cryoballoon PV isolation. While the postoperative AF recurrence was comparable, the postoperative reduction in the cross-sectional PV area was less than that of the conventional method, which may reduce the risk of PV stenosis.

Anatomical information of the lenticulostriate arteries on high-resolution 3D-TOF MRA at 3 T: comparison with 3D-DSA.

PURPOSE: As the lenticulostriate arteries (LSAs) perfuse neurologically important areas, it is necessary to accurately assess the origin and number of the LSAs before surgery. Although three-dimensional time-of-flight MR angiography (3D-TOF MRA) is a non-invasive procedure, it requires high-resolution (HR) images to depict the LSAs with a small diameter. Therefore, we performed 3D-TOF MRA with the maximum HR (HR-MRA) using a 3 T scanner to examine whether a good depiction of the LSAs, equivalent to that of digital subtraction angiography (DSA), could be obtained. METHODS: Our study group comprised 16 consecutive patients who underwent HR-MRA and 3D-DSA. In both studies, we evaluated the localization of the origin from M1, M2, or A1 segments, their number of stems, and depiction. RESULTS: There was no significant difference in the visualization of the LSAs between HR-MRA and 3D-DSA (P values; M1, M2, and A1 = 0.39, 0.69, and 0.69, respectively), and both the number of stems and the localization of the origin of the LSAs corresponded between the two examinations. CONCLUSION: HR-MRA at 3 T can depict the LSA well. It reveals the number of the LSA stems and the LSA origin comparatively with DSA.

Characterization of a Radical SAM Oxygenase for the Ether Crosslinking in Darobactin Biosynthesis.

Darobactin A is a ribosomally synthesized, post-translationally modified peptide (RiPP) with potent and broad-spectrum anti-Gram-negative antibiotic activity. The structure of darobactin A is characterized by an ether and C-C crosslinking. However, the specific mechanism of the crosslink formation, especially the ether crosslink, remains elusive. Here, using in vitro enzyme assays, we demonstrate that both crosslinks are formed by the DarE radical S-adenosylmethionine (SAM) enzyme in an O2-dependent manner. The relevance of the observed activity to darobactin A biosynthesis was demonstrated by proteolytic transformation of the DarE product into darobactin A. Furthermore, DarE assays in the presence of 18O2 or [18O]water demonstrated that the oxygen of the ether crosslink originates from O2 and not from water. These results demonstrate that DarE is a radical SAM enzyme that uses oxygen as a co-substrate in its physiologically relevant function. Since radical SAM enzymes are generally considered to function under anaerobic environments, the discovery of a radical SAM oxygenase represents a significant change in the paradigm and suggests that these radical SAM enzymes function in aerobic cells. Also, the study revealed that DarE catalyzes the formation of three distinct modifications on DarA; ether and C-C crosslinks and α,ß-desaturation. Based on these observations, possible mechanisms of the DarE-catalyzed reactions are discussed.

A 15-year follow-up study of radiofrequency catheter ablation for atrial fibrillation in patients with tachycardia-bradycardia syndrome.

INTRODUCTION: Catheter ablation for atrial fibrillation (AF) in patients with tachycardia-bradycardia syndrome (TBS) can be a major therapeutic option to replace permanent pacemaker implantation (PMI). However, the very long-term outcome of more than 15 years in these patients has not been elucidated. METHODS: From 2002 to 2008, 25 consecutive TBS patients (62 ± 7.9 years old, 68% male) with both AF and symptomatic sinus pauses (>3.0 s) were performed radiofrequency AF ablation. These patients were followed for 15 ± 2.7 years. RESULTS: The median longest sinus pause before the ablation procedure was 6.0 s (4.4-8.0). Following 1.6± 0.8 ablation procedures, 18 (72%) patients remained free from AF. Three (12%) patients died due to noncardiovascular causes, and seven (28%) patients underwent PMI due to symptomatic sinus pause after recurrent AF in five patients and progression of sinus node dysfunction in two patients. The median duration from the first AF ablation to PMI was 6.3 years (range: 9 days to 11.0 years). Five and two patients required PMI more than 5 and 10 years after the first ablation procedure, respectively. CONCLUSION: AF ablation prevented PMI in 72% of TBS patients for a 15-year follow-up. However, in consideration of the long duration of PMI, a continuous careful long-term follow-up was warranted.

Clinical validity of non-contrast-enhanced VI-RADS: prospective study using 3-T MRI with high-gradient magnetic field.

OBJECTIVES: To develop a modified Vesical Imaging Reporting and Data System (VI-RADS) without dynamic contrast-enhanced imaging (DCEI), termed "non-contrast-enhanced VI-RADS (NCE-VI-RADS)", and to assess the additive impact of denoising deep learning reconstruction (dDLR) on NCE-VI-RADS. METHODS: From January 2019 through December 2020, 163 participants who underwent high-gradient 3-T MRI of the bladder were prospectively enrolled. In total, 108 participants with pathologically confirmed bladder cancer by transurethral resection were analyzed. Tumors were evaluated based on VI-RADS (scores 1-5) by two readers independently: an experienced radiologist (reader 1) and a senior radiology resident (reader 2). Conventional VI-RADS assessment included all three imaging types (T2-weighted imaging [T2WI], diffusion-weighted imaging [DWI], and dynamic contrast-enhanced imaging [DCEI]). Also evaluated were NCE-VI-RADS comprising only non-contrast-enhanced imaging types (T2WI and DWI), and "NCE-VI-RADS with dDLR" comprising T2WI processed with dDLR and DWI. All systems were assessed using receiver-operating characteristic curve analysis and simple and/or weighted κ statistics. RESULTS: Muscle invasion was identified in 23/108 participants (21%). Area under the curve (AUC) values for diagnosing muscle invasion were as follows: conventional VI-RADS, 0.94 and 0.91; NCE-VI-RADS, 0.93 and 0.91; and "NCE-VI-RADS with dDLR", 0.96 and 0.93, for readers 1 and 2, respectively. Simple κ statistics indicated substantial agreement for NCE-VI-RADS and almost perfect agreement for conventional VI-RADS and "NCE-VI-RADS with dDLR" between the two readers. CONCLUSION: NCE-VI-RADS achieved predictive accuracy for muscle invasion comparable to that of conventional VI-RADS. Additional use of dDLR improved the diagnostic accuracy of NCE-VI-RADS. KEY POINTS: • Non-contrast-enhanced Vesical Imaging Reporting and Data System (NCE-VI-RADS) was developed to avoid risk related to gadolinium-based contrast agent administration. • NCE-VI-RADS had predictive accuracy for muscle invasion comparable to that of conventional VI-RADS. • The additional use of denoising deep learning reconstruction (dDLR) might further improve the diagnostic accuracy of NCE-VI-RADS.

Transition of the heart rate and atrial premature complex after cryoballoon vs. radiofrequency ablation for paroxysmal atrial fibrillation.

The temporal changes in ambulatory monitoring findings after cryoballoon (CB) ablation of atrial fibrillation (AF) have not been well elucidated. This study aims to compare the details of ambulatory monitoring after CB and radiofrequency catheter (RFC) ablation for AF. Of 724 consecutive AF patients who underwent initial ablation using a CB or RFC, 508 (254 pairs) were selected using propensity score matching. Ambulatory monitoring was performed at 1, 3, 6, 12, 24 and 36 months after the procedure. After 1, 3 and 6 months, the number of total heart beats (THBs) was larger in the CB group than in the RFC group. It gradually decreased and became significantly similar by 12 months after ablation. THBs significantly increased 1, 3, 6 and 12 months after ablation in both the RFC and CB groups and became statistically similar by 24 months after ablation. The atrial premature contraction burden was higher in the RFC group than in the CB group at 3 months after ablation. THB and APC burden after AF ablation were significantly different between the RF and CB groups. THBs returned to statistically similarity by 2 years after ablation in both groups.

Development and performance evaluation of a deep learning lung nodule detection system.

BACKGROUND: Lung cancer is the leading cause of cancer-related deaths throughout the world. Chest computed tomography (CT) is now widely used in the screening and diagnosis of lung cancer due to its effectiveness. Radiologists must identify each small nodule shadow from 3D volume images, which is very burdensome and often results in missed nodules. To address these challenges, we developed a computer-aided detection (CAD) system that automatically detects lung nodules in CT images. METHODS: A total of 1997 chest CT scans were collected for algorithm development. The algorithm was designed using deep learning technology. In addition to evaluating detection performance on various public datasets, its robustness to changes in radiation dose was assessed by a phantom study. To investigate the clinical usefulness of the CAD system, a reader study was conducted with 10 doctors, including inexperienced and expert readers. This study investigated whether the use of the CAD as a second reader could prevent nodular lesions in lungs that require follow-up examinations from being overlooked. Analysis was performed using the Jackknife Free-Response Receiver-Operating Characteristic (JAFROC). RESULTS: The CAD system achieved sensitivity of 0.98/0.96 at 3.1/7.25 false positives per case on two public datasets. Sensitivity did not change within the range of practical doses for a study using a phantom. A second reader study showed that the use of this system significantly improved the detection ability of nodules that could be picked up clinically (p = 0.026). CONCLUSIONS: We developed a deep learning-based CAD system that is robust to imaging conditions. Using this system as a second reader increased detection performance.

Conserved Mechanism of 2'-Phosphorylation-Aided Amide Ligation in Peptidyl Nucleoside Biosynthesis.

Peptidyl nucleoside antifungals, represented by nikkomycins and polyoxins, consist of an unusual six-carbon nucleoside [aminohexuronic acid (AHA)] ligated to a nonproteinogenic amino acid via an amide bond. A recent study suggested that AHA is biosynthesized through cryptic phosphorylation, where a 2'-phosphate is introduced early in the pathway and required to form AHA. However, whether 2'-phosphorylation is necessary for the last step of biosynthesis, the formation of the amide bond between AHA and nonproteinogenic amino acids, remains ambiguous. Here, we address this question with comprehensive in vitro and in vivo characterizations of PolG and NikS, which together provide strong evidence that amide ligation proceeds with 2'-phosphorylated substrates in both pathways. Our results suggest that 2'-phosphorylation is retained for the entirety of both nikkomycin and polyoxin biosynthesis, providing important insights into how cryptic phosphorylation assists with nucleoside natural product biosynthesis.

Mechanism of Reduction of an Aminyl Radical Intermediate in the Radical SAM GTP 3',8-Cyclase MoaA.

The diversity of the reactions catalyzed by radical S-adenosyl-l-methionine (SAM) enzymes is achieved at least in part through the variety of mechanisms to quench their radical intermediates. In the SPASM-twitch family, the largest family of radical SAM enzymes, the radical quenching step is thought to involve an electron transfer to or from an auxiliary 4Fe-4S cluster in or adjacent to the active site. However, experimental demonstration of such functions remains limited. As a representative member of this family, MoaA has one radical SAM cluster ([4Fe-4S]RS) and one auxiliary cluster ([4Fe-4S]AUX), and catalyzes a unique 3',8-cyclization of GTP into 3',8-cyclo-7,8-dihydro-GTP (3',8-cH2GTP) in the molybdenum cofactor (Moco) biosynthesis. Here, we report a mechanistic investigation of the radical quenching step in MoaA, a chemically challenging reduction of 3',8-cyclo-GTP-N7 aminyl radical. We first determined the reduction potentials of [4Fe-4S]RS and [4Fe-4S]AUX as -510 mV and -455 mV, respectively, using a combination of protein film voltammogram (PFV) and electron paramagnetic resonance (EPR) spectroscopy. Subsequent Q-band EPR characterization of 5'-deoxyadenosine C4' radical (5'-dA-C4'•) trapped in the active site revealed isotropic exchange interaction (∼260 MHz) between 5'-dA-C4'• and [4Fe-4S]AUX1+, suggesting that [4Fe-4S]AUX is in the reduced (1+) state during the catalysis. Together with density functional theory (DFT) calculation, we propose that the aminyl radical reduction proceeds through a proton-coupled electron transfer (PCET), where [4Fe-4S]AUX serves as an electron donor and R17 residue acts as a proton donor. These results provide detailed mechanistic insights into the radical quenching step of radical SAM enzyme catalysis.

Prospective Validation of Vesical Imaging-Reporting and Data System Using a Next-Generation Magnetic Resonance Imaging Scanner-Is Denoising Deep Learning Reconstruction Useful?

PURPOSE: The Vesical Imaging Reporting and Data System (VI-RADS) was launched in 2018 to standardize reporting of magnetic resonance imaging for bladder cancer. This study aimed to prospectively validate VI-RADS using a next-generation magnetic resonance imaging scanner and to investigate the usefulness of denoising deep learning reconstruction. MATERIALS AND METHODS: We prospectively enrolled 98 patients who underwent bladder multiparametric magnetic resonance imaging using a next-generation magnetic resonance imaging scanner before transurethral resection of bladder tumor. Tumors were categorized according to VI-RADS, and we ultimately analyzed 68 patients with pathologically confirmed urothelial bladder cancer. We used receiving operating characteristic curve analyses to assess the predictive accuracy of VI-RADS for muscle invasion. Sensitivity, specificity, positive/negative predictive value, accuracy and area under the curve were calculated for different VI-RADS score cutoffs. RESULTS: Muscle invasion was detected in the transurethral resection of bladder tumor specimens of 18 patients (26%). The optimal cutoff value of the VI-RADS score was determined as ≥4 based on the receiver operating curve analyses. The accuracy of diagnosing muscle invasion using a cutoff of VI-RADS ≥4 was 94% (AUC 0.92). Additionally, we assessed the utility of denoising deep learning reconstruction. Combination with denoising deep learning reconstruction significantly improved the AUC of category by T2-weighted imaging, and of the 4 patients who were misdiagnosed by the final VI-RADS score 3 were correctly diagnosed by T2-weighted imaging+denoising deep learning reconstruction. CONCLUSIONS: In this prospective validation study with a next-generation magnetic resonance imaging scanner, VI-RADS showed high predictive accuracy for muscle invasion in patients with bladder cancer before transurethral resection of bladder tumor. Combining T2-weighted imaging with denoising deep learning reconstruction might further improve the diagnostic accuracy of VI-RADS.

PKD1-Associated Arachnoid Cysts in Autosomal Dominant Polycystic Kidney Disease.

OBJECTIVES: the prevalence of intracranial aneurysms and arachnoid cysts is higher in patients with autosomal dominant polycystic kidney disease (ADPKD) than in the general population. A genotype correlation was reported for intracranial aneurysms, but it is unclear for arachnoid cysts. Therefore, the genotype correlation with intracranial aneurysms and arachnoid cysts was investigated in ADPKD. MATERIALS AND METHODS: intracranial aneurysms and arachnoid cysts were screened by magnetic resonance imaging (MRI), and PKD genotypes were examined using next-generation sequencing for 169 patients with ADPKD. RESULTS: PKD1-, PKD2- and no-mutation were identified in 137, 24 and 8 patients, respectively. Intracranial aneurysms and arachnoid cysts were found in 34 and 25 patients, respectively, with no significant difference in frequency. Genotype, sex, estimated glomerular filtration rate and age at ADPKD diagnosis significantly affected the age at brain MRI. The proportional hazard risk analyzed using the age at brain MRI adjusted by these four variables was 5.0-times higher in the PKD1 group than in the PKD2 group for arachnoid cysts (P = 0.0357), but it was not different for intracranial aneurysms (P = 0.1605). Arachnoid cysts were diagnosed earlier in the PKD1 group than in the PKD2 group (54.8 vs 67.7 years, P = 0.0231), but no difference was found for intracranial aneurysms (P = 0.4738) by Kaplan-Meier analysis. CONCLUSIONS: this study demonstrated the correlation between arachnoid cysts and PKD1 mutation. The reported association of arachnoid cysts with advanced renal disease may be due to the common correlation of these factors with PKD1 mutation.

Length Specificity and Polymerization Mechanism of (1,3)-ß-d-Glucan Synthase in Fungal Cell Wall Biosynthesis.

(1,3)-ß-d-Glucan synthase (GS) catalyzes formation of the linear (1,3)-ß-d-glucan in the fungal cell wall and is a target of clinically approved antifungal antibiotics. The catalytic subunit of GS, FKS protein, does not exhibit significant sequence homology to other glycosyltransferases, and thus, significant ambiguity about its catalytic mechanism remains. One of the major technical barriers in studying GS is the absence of activity assay methods that allow characterization of the lengths and amounts of (1,3)-ß-d-glucan due to its poor solubility in water and organic solvents. Here, we report a successful development of a novel GS activity assay based on size-exclusion chromatography coupled with pulsed amperometric detection and radiation counting (SEC-PAD-RC), which allows for the simultaneous characterization of the amount and length of the polymer product. The assay revealed that the purified yeast GS produces glucan with a length of 6550 ± 760 mer, consistent with the reported degree of polymerization of (1,3)-ß-d-glucan isolated from intact cells. Pre-steady state kinetic analysis revealed a highly efficient but rate-determining chain elongation rate of 51.5 ± 9.8 s-1, which represents the first observation of chain elongation by a nucleotide-sugar-dependent polysaccharide synthase. Coupling the SEC-PAD-RC method with substrate analogue mechanistic probes provided the first unambiguous evidence that GS catalyzes non-reducing end polymerization. On the basis of these observations, we propose a detailed model for the catalytic mechanism of GS. The approaches described here can be used to determine the mechanism of catalysis of other polysaccharide synthases.

Mechanism of Rate Acceleration of Radical C-C Bond Formation Reaction by a Radical SAM GTP 3',8-Cyclase.

While the number of characterized radical S-adenosyl-l-methionine (SAM) enzymes is increasing, the roles of these enzymes in radical catalysis remain largely ambiguous. In radical SAM enzymes, the slow radical initiation step kinetically masks the subsequent steps, making it impossible to study the kinetics of radical chemistry. Due to this kinetic masking, it is unknown whether the subsequent radical reactions require rate acceleration by the enzyme active site. Here, we report the first evidence that a radical SAM enzyme MoaA accelerates the radical-mediated C-C bond formation. MoaA catalyzes an unprecedented 3',8-cyclization of GTP into 3',8-cyclo-7,8-dihydro-GTP (3',8-cH2GTP) during the molybdenum cofactor (Moco) biosynthesis. Through a series of EPR and biochemical characterizations, we found that MoaA catalyzes a shunt pathway in which an on-pathway intermediate, GTP C-3' radical, abstracts H-4' atom from (4'R)-5'-deoxyadenosine (5'-dA) to transiently generate 5'-deoxyadenos-4'-yl radical (5'-dA-C4'•) that is subsequently reduced stereospecifically to yield (4'S)-5'-dA. Detailed kinetic characterization of the shunt and the main pathways provided the comprehensive view of MoaA kinetics and determined the rate of the on-pathway 3',8-cyclization step as 2.7 ± 0.7 s-1. Together with DFT calculations, this observation suggested that the 3',8-cyclization by MoaA is accelerated by 6-9 orders of magnitude. Further experimental and theoretical characterizations suggested that the rate acceleration is achieved mainly by constraining the triphosphate and guanine base positions while leaving the ribose flexible, and a transition state stabilization through H-bond and electrostatic interactions with the positively charged R17 residue. This is the first evidence for rate acceleration of radical reactions by a radical SAM enzyme and provides insights into the mechanism by which radical SAM enzymes accelerate radical chemistry.

Effect of Ultra High-Resolution Computed Tomography and Model-Based Iterative Reconstruction on Detectability of Simulated Submillimeter Artery.

OBJECTIVE: To evaluate the effect of ultra high-resolution computed tomography (UHRCT) and model-based iterative reconstruction (MBIR) on the detectability of simulated submillimeter artery. METHODS: A small vessel phantom ranging from 0.4 to 2.0 mm in diameter and edge phantoms of low to high attenuation values were scanned by UHRCT (super-high-resolution mode and normal-resolution-mode) and conventional CT, and data were reconstructed by MBIR and filtered back projection (FBP). Vessel detectability was assessed subjectively and the effective size at which 50% of response was achieved (ES50 [mm]) was calculated. Modulation transfer function (MTF) was calculated by an edge spread function method. RESULTS: ES50 of super high-resolution mode (0.36 mm for MBIR and 0.50 mm for FBP) was significantly smaller than those of normal-resolution mode (P < 0.01). In the MTF analysis, the MTF of MBIR improved as the edge phantom attenuation increased, whereas that of FBP was stable. CONCLUSIONS: Both UHRCT and MBIR are effective for the detectability of simulated submillimeter artery.

Compositional dependence of the apatite formation ability of Ti-Zr alloys designed for hard tissue reconstruction.

Ti-Zr alloys are expected to be novel biomaterials with low stress shielding owing to their lower Young's moduli than pure Ti. The drawback of metallic biomaterials is that their bone-bonding abilities are relatively low. NaOH and heat treatments have been performed to provide Ti-50Zr with apatite-forming ability in the body environment, which is essential for bone bonding. However, the systematic compositional dependence of apatite formation has not been revealed. In the present study, NaOH treatment of Ti-Zr alloys with various compositions and bone-bonding abilities was assessed in vitro by apatite formation in simulated body fluid (SBF). The corrosion current density in NaOH aqueous solution and the amount of Na incorporated into the surface tended to decrease with increasing Zr content. The apatite-forming ability of the treated alloy significantly decreased when the Zr content was ≥60 atom%. This phenomenon is attributed to the (1) low OH content on the surface, (2) low Na incorporation into the treated alloy surface, which enhances apatite formation, and (3) low ability of P adsorption to the Ti-Zr alloy in SBF following Ca adsorption to trigger apatite nucleation. Although the adhesion of the titanate/zirconate layer formed on the surfaces to the substrates increased as Zr content increased, the adhesion between the apatite and the substrate was still low.

Characterization of Acyl Carrier Protein-Dependent Glycosyltransferase in Mitomycin C Biosynthesis.

Mitomycins make up a group of antitumor natural products that are biosynthesized from aminohydroxybenzoic acid (AHBA) and N-acetylglucosamine (GlcNAc). While the biosynthetic gene cluster was reported two decades ago, the mechanism by which the two building blocks, AHBA and GlcNAc, are coupled during biosynthesis remained uncharacterized. Here we report evidence that AHBA is first loaded onto an MmcB acyl carrier protein (ACP) by a MitE acyl ACP synthetase, followed by a transfer of GlcNAc from UDP-GlcNAc by MitB. The results suggest that the early steps of mitomycin biosynthesis proceed via intermediates linked to MmcB.