Cloning, DNA sequence, and expression of flagellins from high and low virulence strains of Edwardsiella tarda and their macrophage-stimulating activities.

Edwardsiella tarda is a causative pathogen of edwardsiellosis in fish. Our previous studies on high (NUF251) and low (NUF194) virulent strains of E. tarda demonstrated that NUF251 strain induced significantly higher levels of NO and TNF-α from fish and mouse macrophages than NUF194 strain. Subsequent studies suggested that a flagellin-like protein secreted from E. tarda might be a responsible factor for the macrophage-stimulating activities. To evaluate the activities of flagellins of E. tarda, in this study, the flagellin genes of NUF251 and NUF194 strains were isolated by PCR and cloned into pQE-30 and pCold I expression vectors, and then the recombinant flagellins of two strains were overexpressed in E. coli JM109 and pG-Tf/BL21, respectively. The molecular weight of the purified recombinant flagellins of NUF251 and NUF194 strains were estimated to be 45 kDa and 37 kDa, respectively on SDS-PAGE analysis. Referring the three-dimensional structure of Salmonella flagellin, which has been reported to have 4 domains (D0, D1, D2, and D3), high sequence homology between two flagellins of E. tarda was observed at conservative domain (D0 and D1) regions, whereas the sequences equivalent to D2 and D3 domains were different, and even equivalent to 57 amino acids were deleted in NUF194. Both recombinant flagellins induced NO production, mRNA expression level of inducible NO synthase (iNOS), and intercellular ROS production in mouse macrophage cell line RAW264.7 cells. Also, the secretion of TNF-α and its mRNA expression level were increased by treatment of both recombinant flagellins. These results indicate that the recombinant flagellins from different virulent E. tarda strains can stimulate macrophages with nearly equal levels as judged by the parameters tested, even though they are differences in the structure and molecular weight, suggesting that conservative D0 and D1 domains are sufficient structural elements for the recombinant flagellins to induce a certain level of macrophage-stimulation in vitro. Further studies are necessary focusing on the role of D2 and D3 domain regions of the recombinant flagellins as macrophage-stimulating agent as well as their influence on host immune system in vivo.

Wheat germ agglutinin affinity chromatography enrichment and glyco-proteomic characterization of tetrodotoxin-binding proteins from the plasma of cultured tiger pufferfish (Takifugu rubripes).

Efficient enrichment of tetrodotoxin (TTX)-binding proteins from the plasma of cultured tiger pufferfish (Takifugu rubripes) was achieved by ammonium sulfate fractionation and wheat germ agglutinin (WGA) affinity chromatography. The enrichment efficiency was validated by ultrafiltration-LC/MS-based TTX-binding assay and proteomics. Major proteins in the WGA-bound fraction were identified as isoform X1 (125 kDa) and X2 variants (88 and 79 kDa) derived from pufferfish saxitoxin and tetrodotoxin-binding protein (PSTBP) 1-like gene (LOC101075943). The 125-kDa X1 protein was found to be a novel member of the lipocalin family, having three tandemly repeated domains. X2 variants, X2α and X2ß, were estimated to have two domains, and X2ß is structurally related to Takifugu pardalis PSTBP2 in their domain type and arrangement. Among 11 potential N-glycosylation sites in the X2 precursor, 5 N-glycosylated Asn residues (N55, N89, N244, N308, and N449) were empirically determined. Structural relationships among PSTBP homologs and complexity of their proteoforms are discussed.

Identification and Characterization of the Larval Settlement Pheromone Protein Components in Adult Shells of Crassostrea gigas: A Novel Function of Shell Matrix Proteins.

The global decline of natural oyster populations emphasizes the need to improve our understanding of their biology. Understanding the role of chemical cues from conspecifics on how oysters occupy appropriate substrata is crucial to learning about their evolution, population dynamics, and chemical communication. Here, a novel role of a macromolecular assembly of shell matrix proteins which act as Crassostrea gigas Settlement Pheromone Protein Components in adult shells is demonstrated as the biological cue responsible for gregarious settlement on conspecifics. A bioassay-guided fractionation approach aided by biochemical and molecular analyses reveals that Gigasin-6 isoform X1 and/or X2 isolated from adult shells is the major inducing cue for larval settlement and may also play a role in postlarva-larva settlement interactions. Other isolated Stains-all-stainable acidic proteins may function as a co-factor and a scaffold/structural framework for other matrix proteins to anchor within this assembly and provide protection. Notably, conspecific cue-mediated larval settlement induction in C. gigas presents a complex system that requires an interplay of different glycans, disulfide bonds, amino acid groups, and phosphorylation crosstalk for recognition. These results may find application in the development of oyster aquacultures which could help recover declining marine species and as targets of anti-fouling agents.

Regulatory Role of Sugars on the Settlement Inducing Activity of a Conspecific Cue in Pacific Oyster Crassostrea gigas.

This study evaluated the larval settlement inducing effect of sugars and a conspecific cue from adult shell extract of Crassostrea gigas. To understand how the presence of different chemical cues regulate settlement behavior, oyster larvae were exposed to 12 types of sugars, shell extract-coated and non-coated surfaces, and under varied sugar exposure times. Lectin-glycan interaction effects on settlement and its localization on oyster larval tissues were investigated. The results showed that the conspecific cue elicited a positive concentration dependent settlement inducing trend. Sugars in the absence of a conspecific cue, C. gigas adult shell extract, did not promote settlement. Whereas, in the presence of the cue, showed varied effects, most of which were found inhibitory at different concentrations. Sugar treated larvae exposed for 2 h showed significant settlement inhibition in the presence of a conspecific cue. Neu5Ac, as well as GlcNAc sugars, showed a similar interaction trend with wheat germ agglutinin (WGA) lectin. WGA-FITC conjugate showed positive binding on the foot, velum, and mantle when exposed to GlcNAc sugars. This study suggests that a WGA lectin-like receptor and its endogenous ligand are both found in the larval chemoreceptors and the shell Ethylenediaminetetraacetic acid (EDTA) extract that may complementarily work together to allow the oyster larva greater selectivity during site selection.

A bacterial polysaccharide biosynthesis-related gene inversely regulates larval settlement and metamorphosis of Mytilus coruscus.

Larval settlement and metamorphosis is essential for the development of marine invertebrates. Although polysaccharides are involved in larval settlement and metamorphosis of Mytilus coruscus, the molecular basis of polysaccharides underlying this progression remains largely unknown. Here, the roles of the polysaccharide biosynthesis-related gene 01912 of Pseudoalteromonas marina ECSMB14103 in the regulation of larval settlement and metamorphosis were examined by gene-knockout technique. Compared with biofilms (BFs) of the wild-type P. marina, Δ01912 BFs with a higher colanic acid (CA) content showed a higher inducing activity on larval settlement and metamorphosis. Deletion of the 01912 gene caused an increase in c-di-GMP levels, accompanied by a decrease in the motility, an increase in cell aggregation, and overproduction of CA. Thus, the bacterial polysaccharide biosynthesis-related gene 01912 may regulate mussel settlement by producing CA via the coordination of c-di-GMP. This work provides a deeper insight into the molecular mechanism of polysaccharides in modulating mussel settlement.

Clinical results of transarterial embolization for post-partum hemorrhage in 62 patients.

AIM: The pathology of post-partum hemorrhage (PPH) differs depending on its cause, background and timing of bleeding, and the effectiveness of transarterial embolization (TAE) is thought to vary based on these characteristics. The aim of this study is to evaluate the treatment outcomes of TAE for PPH. METHODS: Technical success, initial clinical success (hemostasis without repeat TAE or surgical treatment after initial TAE) and final clinical success (hemostasis with or without repeat TAE, but without surgical treatment) were assessed in 62 Japanese patients. Factors affecting final clinical success were analyzed using univariate analysis. Values of P < 0.05 were considered statistically significant. Further, the clinical course and factors associated with rebleeding, return of menstruation and fertility, and complications of TAE were assessed. RESULTS: Final clinical success rate was significantly lower in cases with obstetrical disseminated intravascular coagulation (DIC) or the International Society on Thrombosis and Hemostasis (ISTH) DIC (P = 0.01, 0.03). Rebleeding (n = 9, 14.5%) was more common in patients with retained products of conception (RPOC) (P = 0.006). On long-term follow-up in 23 patients, return of menstruation was confirmed in 17 (73.9%) of these patients. Subsequent pregnancy was confirmed in seven patients (30.4%). TAE-related complications were seen in 6 patients (9.0%). There were no maternal deaths. CONCLUSIONS: Obstetrical and ISTH DIC reduced the success rate of TAE for PPH (P = 0.01, 0.03). Rebleeding, which is observed significantly more frequently in PPH caused by RPOC (P = 0.006), can be effectively treated by repeat TAE.

The Flagellar Gene Regulates Biofilm Formation and Mussel Larval Settlement and Metamorphosis.

Biofilms are critical components of most marine systems and provide biochemical cues that can significantly impact overall community composition. Although progress has been made in the bacteria-animal interaction, the molecular basis of modulation of settlement and metamorphosis in most marine animals by bacteria is poorly understood. Here, Pseudoalteromonas marina showing inducing activity on mussel settlement and metamorphosis was chosen as a model to clarify the mechanism that regulates the bacteria-mussel interaction. We constructed a flagellin synthetic protein gene fliP deletion mutant of P. marina and checked whether deficiency of fliP gene will impact inducing activity, motility, and extracellular polymeric substances of biofilms. Furthermore, we examined the effect of flagellar proteins extracted from bacteria on larval settlement and metamorphosis. The deletion of the fliP gene caused the loss of the flagella structure and motility of the ∆fliP strain. Deficiency of the fliP gene promoted the biofilm formation and changed biofilm matrix by reducing ß-polysaccharides and increasing extracellular proteins and finally reduced biofilm-inducing activities. Flagellar protein extract promoted mussel metamorphosis, and ∆fliP biofilms combined with additional flagellar proteins induced similar settlement and metamorphosis rate compared to that of the wild-type strain. These findings provide novel insight on the molecular interactions between bacteria and mussels.

Experimental verification of factors influencing calcium salt formation based on a survey of the development of ceftriaxone-induced gallstone-related disorder.

Ceftriaxone (CTRX) forms salts with calcium (Ca) in the gall bladder and bile duct, and induces the formation of gallstones. In this study, factors of CTRX-induced gallstone formation were extracted from the results of a retrospective survey using the Japanese Adverse Drug Event Report (JADER), and the causal relationship between the factors and gallstone formation was investigated. From JADER, 136 patients who developed 'gallstone-related disorder' with CTRX as a suspected drug were extracted. The incidence of gallstone-induced adverse effects was high in patients treated with CTRX at a dose exceeding the normal daily dose and in children younger than 10 years old, suggesting that CTRX at a high level is a factor for gallstone formation. Thus, after mixing CTRX and Ca2+ at different concentrations under different pH condition, the number of particles in the solutions was measured using a Coulter counter. As a result, the number of minute particles significantly increased at all pH values when Ca2+ and CTRX were mixed at a concentration of 10 mEq/L or higher and 1.5 g/L or higher, respectively. At pH 6.5 or 7.0, visible crystals were detected when 25 mEq/L of Ca2+ and 2.0 g/L of CTRX were mixed. Based on these findings, attention should be sufficiently paid to the development of 'gallstone-related disorder' in pediatric patients and in patients treated with CTRX at a dose exceeding the normal dose. Furthermore, gallstone formation and growth may be promoted when CTRX and Ca2+ coexist at high concentrations under low pH conditions.

An ɑ2-adrenergic receptor is involved in larval metamorphosis in the mussel, Mytilus coruscus.

Metamorphosis is crucial in the life-cycle transition between the larval and juvenile stages of marine invertebrates. Although a number of agonists and antagonists of the adrenergic receptor (AR) are known to regulate larval metamorphosis in Mytilus coruscus (Mc), the molecular basis of the modulation of larval metamorphosis by the AR gene in this species remains elusive. Herein, the role of the AR gene in M. coruscus larval metamorphosis using the RNA interference technique was examined. The Mcα2AR transcript was observed to be present during the entire process of larval development and its level in the post-larvae was significantly increased compared to that in the pediveligers. Mcα2AR-knockdown resulted in a substantial reduction in the abundance of the Mcα2AR transcript and significantly inhibited the metamorphosis of M. coruscus larvae. These findings provide new insights into the molecular basis of modulation of larval metamorphosis in M. coruscus by the AR gene.

Purification and Characterization of Cathepsin B from the Muscle of Horse Mackerel Trachurus japonicus.

An endogenous protease in fish muscle, cathepsin B, was partially purified and characterized from horse mackerel meat. On SDS-PAGE of the purified enzyme under reducing conditions, main protein bands were detected at 28 and 6 kDa and their respective N-terminal sequences showed high homology to heavy and light chains of cathepsin B from other species. This suggested that horse mackerel cathepsin B formed two-chain forms, similar to mammalian cathepsin Bs. Optimum pH and temperature of the enzyme were 5.0 and 50 °C, respectively. A partial cDNA encoding the amino acid sequence of 215 residues for horse mackerel cathepsin B was obtained by RT-PCR and cloned. The deduced amino acid sequence contains a part of light and heavy chains of cathepsin B. The active sites and an N-glycosylation site were conserved across species. We also confirmed that the modori phenomenon was avoided by CA-074, a specific inhibitor for cathepsin B. Therefore, our results suggest that natural cysteine protease inhibitor(s), such as oryzacystatin derived from rice, can apply to thermal-gel processing of horse mackerel to avoid the modori phenomenon. Meanwhile, this endogenous protease may be used for food processing, such as weaning meal and food for the elderly.

Transcriptional regulation of the Japanese flounder Cu,Zn-SOD (Jfsod1) gene in RAW264.7 cells during oxidative stress caused by causative bacteria of edwardsiellosis.

Edwardsiellosis is one of the most important bacterial diseases in fish, sometimes causing extensive economic losses in the aquaculture industry. Our previous studies demonstrated that the Cu,Zn-SOD (sod1) activity has significantly increased in Japanese flounder, Paralichthys olivaceus, hepatopancreas infected by causative bacteria of edwardsiellosis Edwardsiella tarda NUF251. In this study, NUF251 stimulated intracellular superoxide radical production in mouse macrophage RAW264.7 cells, which was reduced by N-acetylcysteine. This result suggests that NUF251 infection causes oxidative stress. To evaluate the regulatory mechanism of Jfsod1 at transcriptional levels under oxidative stress induced by NUF251 infection, we cloned and determined the nucleotide sequence (1124 bp) of the 5'-flanking region of the Jfsod1 gene. The sequence analysis demonstrated that the binding sites for the transcription factors C/EBPα and NF-IL6 involved in the transcriptional regulation of the mammalian sod1 gene existed. We constructed a luciferase reporter system with the 5'-flanking region (-1124/-1) of the Jfsod1 gene, and a highly increased transcriptional activity of the region was observed in NUF251-infected RAW264.7 cells. Further studies using several mutants indicated that deletion of the recognition region of NF-IL6 (-272/-132) resulted in a significant decrease in the transcriptional activity of the Jfsod1 gene in NUF251-infected RAW264.7 cells. In particular, the binding site (-202/-194) for NF-IL6 might play a major role in upregulating the transcriptional activity of the 5'-flanking region of the Jfsod1 gene in response to oxidative stress induced by NUF251 infection. These results could be provided a new insight to understand the pathogenic mechanism of causative bacteria of edwardsiellosis.

Single-Fraction Palliative High-Dose-Rate Brachytherapy for Symptom Management in a 97-Year-Old Patient With Dementia.

Background: Delivering cancer treatment to elderly patients with dementia is often challenging. We describe performing palliative surface mold brachytherapy (SMBT) in an elderly patient with advanced dementia for pain control using music therapy to assist with agitation. Case Description: The patient was a 97-year-old Japanese woman with advanced dementia. Exudate was observed from her tumor, and she complained of Grade 2 severity pain using Support team assessment schedule (STAS), especially when undergoing would dressings. Given her advanced dementia, she was not considered a candidate for radical surgery or external beam radiotherapy. We instead treated her with high-dose-rate (HDR) SMBT. Due to her advanced dementia associated with agitation, she could not maintain her position. She was able to remain calm while listening to traditional Japanese enka music, which enables our team to complete her radiation without using anesthetics or sedating analgesics. Her localized pain severity decreased ≤21 days and the exudate fluid disappeared ≤63 days after HDR-SMBT. Her tumor was locally controlled until her death from intercurrent disease 1 year after HDR-SMBT. Discussion: Single fraction palliative HDR-SMBT was useful for successful treatment of skin cancer in an elderly patient. Traditional Japanese music helped reduce her agitation to complete HDR-SMBT. For elderly patients with agitation associated with dementia, we should consider using music and music therapy to facilitate radiation therapy.

Artificial intelligence-aided lytic spinal bone metastasis classification on CT scans.

PURPOSE: Spinal bone metastases directly affect quality of life, and patients with lytic-dominant lesions are at high risk for neurological symptoms and fractures. To detect and classify lytic spinal bone metastasis using routine computed tomography (CT) scans, we developed a deep learning (DL)-based computer-aided detection (CAD) system. METHODS: We retrospectively analyzed 2125 diagnostic and radiotherapeutic CT images of 79 patients. Images annotated as tumor (positive) or not (negative) were randomized into training (1782 images) and test (343 images) datasets. YOLOv5m architecture was used to detect vertebra on whole CT scans. InceptionV3 architecture with the transfer-learning technique was used to classify the presence/absence of lytic lesions on CT images showing the presence of vertebra. The DL models were evaluated via fivefold cross-validation. For vertebra detection, bounding box accuracy was estimated using intersection over union (IoU). We evaluated the area under the curve (AUC) of a receiver operating characteristic curve to classify lesions. Moreover, we determined the accuracy, precision, recall, and F1 score. We used the gradient-weighted class activation mapping (Grad-CAM) technique for visual interpretation. RESULTS: The computation time was 0.44 s per image. The average IoU value of the predicted vertebra was 0.923 ± 0.052 (0.684-1.000) for test datasets. In the binary classification task, the accuracy, precision, recall, F1-score, and AUC value for test datasets were 0.872, 0.948, 0.741, 0.832, and 0.941, respectively. Heat maps constructed using the Grad-CAM technique were consistent with the location of lytic lesions. CONCLUSION: Our artificial intelligence-aided CAD system using two DL models could rapidly identify vertebra bone from whole CT images and detect lytic spinal bone metastasis, although further evaluation of diagnostic accuracy is required with a larger sample size.

The Dosimetric Analysis of Duodenal and Intestinal Toxicity After a Curative Dose Re-irradiation Using the Intensity-Modulated Radiotherapy for Abdominopelvic Lymph Node Lesions.

INTRODUCTION: This study aimed to examine the influence of dosimetric factors on gastrointestinal toxicity after radical re-irradiation for lymph node recurrence in the abdominopelvic region using a composite plan. METHODS: Between January 2008 and March 2017, 33 patients underwent radical re-irradiation for lymph node recurrence in the abdominopelvic region with a complete overlap with previous radiation therapy (RT) with the median prescription dose of the second RT of 71.7 Gy10. Re-irradiation planning protocol for target volume and organs at risk (OARs) (duodenum, small and large intestines) was decided as follows: more than equal to 97% of the prescription dose was administered to the D95 (percentage of the minimum dose that covered 95% of the target volume) of planning target volume (PTV); minimal dose to the maximally irradiated doses delivered to 1cc [D1 cc] and 5cc [D5 cc] of OARs was set below 70 Gy3 and 50 Gy3, respectively; and D1 cc and D5 cc in the cumulative plans to OARs were 120 Gy3 and 100 Gy3. Kaplan-Meier analyses were performed to evaluate overall survival (OS) and univariate log-rank and multivariate Cox proportional hazards model analyses were performed to explore predictive factors. Using dose summation of the first and re-irradiation plans, we conducted a dosimetric analysis for grade ≥ 3 toxicities of the duodenum and intestine. RESULTS: With a median follow-up of 18 months, the two-year OS rate was 45.5%. The number of RT fields (localized or multiple) was a significant predisposing factor for OS rate with a hazard ratio of 0.23 (95% confidence interval 0.07-0.73). The two-year OS of the patients with a localized RT field was 63.6% and 9.1% for multiple RT fields (p= 0.00007). Four patients experienced grade ≥3 gastrointestinal toxicity related to re-irradiation (4/33=12.1%). We could not find any predisposing dosimetric value in the comparisons with and without toxicity. CONCLUSIONS: The dose constraints presented in this study are relatively low rates of toxicity, which may be useful when planning re-irradiation. Especially, for the patients who could be treated with localized RT field, radical re-irradiation with a high curative dose is a good option. No dosimetric predisposing factor was found for radical re-irradiation of abdominopelvic lesions in the composite plan.

Identification of a Tissue Inhibitor of Metalloproteinase-2: An Endogenous Inhibitor of Modori-Inducing Insoluble Metalloproteinase from Yellowtail Seriola quinqueradiata Muscle.

The existence of an endogenous protease inhibitor (EPI) was expected from the comparison of the gel properties between washed and nonwashed yellowtail surimi gels. A possible candidate, tissue inhibitor of metalloproteinase-2 (TIMP-2), was partially purified from the soluble fraction of yellowtail muscle, and an 18 kDa protein band was detected by sodium dodecyl-sulfate polyacrylamide gel electrophoresis (SDS-PAGE) under nonreducing conditions and western blot analysis. Its N-terminal amino acid sequence was determined as XSXSPAHPQQAF, with high homology to TIMP-2 from other fish species, suggesting that it was identified as yellowtail TIMP-2. Subsequently, full-length cDNA of two isoforms (TIMP-2a and TIMP-2b) was successfully cloned from yellowtail muscle. The N-terminal sequence of purified TIMP-2 completely corresponded to TIMP-2b. When the surimi gel quality decreased after spawning, the mRNA expression of TIMP-2b also decreased. Human TIMP-2 could inhibit autolysis of myofibrillar proteins from yellowtail muscle. Thus, TIMP-2b was considered the major EPI of the modori-inducing insoluble metalloproteinase in yellowtail muscle.

Impact of different enzymes on biofilm formation and mussel settlement.

Enzymes have been known to impact the biofilm forming capacity. However, how the enzymes mediate the biofilm formation and macrofouling remains little known. Here, we investigated the effects of the three kinds of proteases, four kinds of glycosidases and one kind of lipase on the detachment of biofilms of Shewanella marisflavi ECSMB14101, identified biofilm total proteins response to enzyme treatments, and then tested the effects of biofilms treated with enzymes on the settlement of the mussel Mytilus coruscus plantigrades. The results showed that the cell density of bacteria in biofilms formed at different initial bacterial density were noticeably reduced after treating with all tested enzymes, and Neutrase and α-Amylase exhibited best removing efficiency of > 90%. Bacterial total proteins in S. marisflavi biofilm noticeably reduced or disappeared after treated by Alcalase. For the settlements of the mussel M. coruscus plantigrades, inducing capacities of S. marisflavi biofilm were noticeably suppressed and downregulation was > 75% at the initial density of 5 × 106 cells/cm2. Thus, the tested enzymes could effectively remove the adhered bacterial cell, inhibit the biofilm formation and finally suppress the mussel settlement. Our findings extend novel knowledge to developing eco-friendly approach to control micro- and macro-fouling.

Functional dissection of the N-terminal degron of human thymidylate synthase.

Human thymidylate synthase (hTS; EC 2.1.1.45) is one of a small group of proteasomal substrates whose intracellular degradation occurs in a ubiquitin-independent manner. Previous studies have shown that proteolytic breakdown of the hTS polypeptide is directed by an intrinsically disordered 27-residue domain at the N-terminal end of the molecule. This domain, in co-operation with an α-helix spanning amino acids 31-45, functions as a degron, in that it has the ability to destabilize a heterologous polypeptide to which it is attached. In the present study, we provide evidence indicating that it is the 26S isoform of the proteasome that is responsible for intracellular degradation of the hTS polypeptide. In addition, we have used targeted in vitro mutagenesis to show that an Arg-Arg motif at residues 10-11 is required for proteolysis, an observation that was confirmed by functional analysis of the TS N-terminus from other mammalian species. The effects of stabilizing mutations on hTS degradation are maintained when the enzyme is provided with an alternative means of proteasome association; thus such mutations perturb one or more post-docking steps in the degradation pathway. Surprisingly, deletion mutants missing large segments of the disordered domain still function as proteasomal substrates; however, degradation of such mutants occurs by a mechanism that is distinct from that for the wild-type protein. Taken together, our results provide information on the roles of specific subregions within the intrinsically disordered N-terminal domain of hTS in regulation of degradation, leading to a deeper understanding of mechanisms underlying the ubiquitin-independent proteasomal degradation pathway.

[Evaluation of Fat Suppression Effect and SNR Using Pixel Shift Method of Fat Suppression Images with CHESS and IDEAL in Head and Neck MR Imaging].

PURPOSE: This study aims to evaluate the fat suppression effect on images of the head and neck region using chemical shift selective (CHESS), and iterative decomposition of water and fat with echo asymmetry and least-squares estimation (IDEAL). METHOD: A self-made phantom containing oil around the simulated bone marrow and muscle was scanned. The signal-to-noise ratio (SNR) was calculated using the National Electrical Manufacturers Association (NEMA) subtraction and pixel shift methods. Thereafter, the fat suppression effect and SNR were calculated in clinical images using the pixel shift method. RESULT: In both phantom and clinical images, the fat suppression effect was higher using IDEAL. In addition, the SNR of the NEMA subtraction method and the pixel shift method in phantom images was higher in the simulated bone marrow than in the simulated muscle. The SNR of the vertebral body was higher than that of the tongue in the clinical images using IDEAL, and the same tendency was observed in the phantom image evaluation. However, there was a difference in SNR between the phantom and clinical images. CONCLUSION: In the head and neck region, fat-suppressed images using IDEAL showed the same higher fat-suppressing effect as that in a previous study. The SNR for the phantom and the clinical images was different. The SNR calculated using the pixel shift method for the phantom images with IDEAL and the clinical images showed the same tendency. Although there is a difference between the SNRs of phantom and clinical images calculated by the pixel shift method, it is suggested that the method can be used to compare the SNR between tissues such as the vertebral body and the tongue.

The Combination of Panoramic Imaging and Waters' Projection Contributes to the Diagnosis of Odontogenic Maxillary Sinusitis.

The purpose of this study was to evaluate the usefulness of adding Waters' projection to panoramic imaging compared with panoramic imaging or Waters' projection alone. Maxillary sinusitis in 106 patients (206 sinuses) was retrospectively assessed with panoramic imaging, Waters' projection, and computed tomography imaging by two oral radiologists. The diagnostic performance was assessed with computed tomography imaging as the gold standard. Receiver operating characteristic curves and area under the curve values were obtained. Inter- and intra-observer agreement was quantified using weighted kappa coefficients. Observer A performed the same procedure twice (A1 and A2 for the first and second observations, respectively). The accuracies of observers A1, B, and A2 with combination imaging were 0.699, 0.636, and 0.718, respectively. Their area under the curve values with combination imaging were 0.746, 0.640, and 0.771, respectively. Inter-observer agreement was good for Waters' projection (κ, 0.650), and poor for panoramic imaging (κ, 0281). Intra-observer agreement was good for Waters' projection (κ, 0.752), and moderate for panoramic imaging (κ, 0.597). Panoramic imaging was equivalent to Waters' projection for diagnosing maxillary sinusitis. Combination imaging comprising panoramic imaging and Waters' projection can contribute to the diagnosis of odontogenic maxillary sinusitis because of its high sensitivity.

A Conserved Histone H3-H4 Interface Regulates DNA Damage Tolerance and Homologous Recombination during the Recovery from Replication Stress.

In eukaryotes, genomic DNA is packaged into nucleosomes, which are the basal components coordinating both the structures and functions of chromatin. In this study, we screened a collection of mutations for histone H3/H4 mutants in Saccharomyces cerevisiae that affect the DNA damage sensitivity of DNA damage tolerance (DDT)-deficient cells. We identified a class of histone H3/H4 mutations that suppress methyl methanesulfonate (MMS) sensitivity of DDT-deficient cells (referred to here as the histone SDD mutations), which likely cluster on a specific H3-H4 interface of the nucleosomes. The histone SDD mutations did not suppress the MMS sensitivity of DDT-deficient cells in the absence of Rad51, indicating that homologous recombination (HR) is responsible for DNA damage resistance. Furthermore, the histone SDD mutants showed reduced levels of PCNA ubiquitination after exposure to MMS or UV irradiation, consistent with decreased MMS-induced mutagenesis relative to that of wild-type cells. We also found that histone SDD mutants lacking the INO80 chromatin remodeler impair HR-dependent recovery from MMS-induced replication arrest, resulting in defective S-phase progression and increased Rad52 foci. Taken together, our data provide novel insights into nucleosome functions, which link INO80-dependent chromatin remodeling to the regulation of DDT and HR during the recovery from replication blockage.