[Rupture of Artificial Blood Vessel by a Sternal Wire:Report of a Case].

Formation of a pseudoaneurysm due to blood leakage from the anastomotic site of the vascular graft in large-diameter vessels is often seen, but formation of a pseudoaneurysm from the non-anastomotic site is extremely rare. A 68-year-old woman presented with a history of double valve replacement for combined valvular disease at 37 years old and hemiarch replacement for thoracic aortic dilatation at 65 years old. She visited the emergency room with a 2-week history of chest pain. Contrast-enhanced computed tomography (CT) revealed a 5-cm-diameter pseudoaneurysm and extravasation from the ascending aorta, so emergency surgery was performed. Around the ascending aorta area, we confirmed bleeding from a 5-mm dehiscence in the non-anastomotic part of the graft prosthesis, so hemostasis was performed with a cross-stitch mattress suture over a felt strip. Initially, the cause of the pseudoaneurysm was unknown, but re-examination of CT images from after the previous hemiarch replacement confirmed contact between the sternal wire and graft prosthesis. The wire was thus considered to have caused damage and bleeding. The patient was discharged from the hospital with a good postoperative course and is being followed-up in the outpatient department.

A blurring correction method suitable to analyze quantitative x-ray images derived from energy-resolving photon counting detector.

Objective.The purpose of this study is to propose a novel blurring correction method that enables accurate quantitative analysis of the object edge when using energy-resolving photon counting detectors (ERPCDs). Although the ERPCDs have the ability to generate various quantitative analysis techniques, such as the derivations of effective atomic number (Zeff) and bone mineral density values, at the object edge in these quantitative images, accurate quantitative information cannot be obtained. This is because image blurring prevents the gathering of accurate primary x-ray attenuation information.Approach.We developed the following procedure for blurring correction. A 5 × 5 pixels masking region was set as the processing area, and the pixels affected by blurring were extracted from the analysis of pixel value distribution. The blurred pixel values were then corrected to the proper values estimated by analyzing minimum and/or maximum values in the set mask area. The suitability of our correction method was verified by a simulation study and an experiment using a prototype ERPCD.Main results. WhenZeffimage of aluminum objects (Zeff= 13) were analyzed without applying our correction method, regardless of raw data or correction data applying a conventional edge enhancement method, the properZeffvalues could not be derived for the object edge. In contrast, when applying our correction method, 82% of pixels affected by blurring were corrected and the properZeffvalues were calculated for those pixels. As a result of investigating the applicability limits of our method through simulation, it was proven that it works effectively for objects with 4 × 4 pixels or more.Significance. Our method is effective in correcting image blurring when the quantitative image is calculated based on multiple images. It will become an in-demand technology for putting a quantitative diagnosis into actual medical examinations.

Performance of elastic x-ray shield made by embedding Bi2 O3 particles in porous polyurethane.

BACKGROUND: Many healthcare institutions have guidelines concerning the usage of protective procedures, and various x-ray shields have been used to reduce unwanted radiation exposure to medical staff and patients when using x-rays. Most x-ray shields are in the form of sheets and lack elasticity, which limits their effectiveness in shielding areas with movement, such as the thyroid. To overcome this limitation, we have developed an innovative elastic x-ray shield. PURPOSE: The purpose of this study is to explain the methodology for developing and evaluating a novel elastic x-ray shield with sufficient x-ray shielding ability. Furthermore, valuable knowledge and evaluation indices are derived to assess our shield's performance. METHODS: Our x-ray shield was developed through a process of embedding Bi2 O3 particles into porous polyurethane. Porous polyurethane with a thickness of 10 mm was dipped into a solution of water, metal particles, and chemical agents. Then, it was air-dried to fix the metal particles in the porous polyurethane. Thirteen investigational x-ray shields were fabricated, in which Bi2 O3 particles at various mass thicknesses (ranging from 585 to 2493 g/m2 ) were embedded. To determine the performance of the shielding material, three criteria were evaluated: (1) Dose Reduction Factor ( D R F $DRF$ ), measured using inverse broad beam geometry; (2) uniformity, evaluated from the standard deviation ( S D $SD$ ) of the x-ray image obtained using a clinical x-ray imaging detector; and (3) elasticity, evaluated by a compression test. RESULTS: The elastic shield with small pores, containing 1200 g/m2 of the metal element (Bi), exhibited a well-balanced performance. The D R F $DRF$ was approximately 80% for 70 kV diagnostic x-rays. This shield's elasticity was -0.62 N/mm, a loss of only 30% when compared to porous polyurethane without metal. Although the non-uniformity of the x-ray shield leads to poor shielding ability, it was found that the decrease in the shielding ability can be limited to a maximum of 6% when the shield is manufactured so that the S D $SD$ of the x-ray image of the shield is less than 10%. CONCLUSIONS: It was verified that an elastic x-ray shield that offers an appropriate reduction in radiation exposure can be produced by embedding Bi2 O3 particles into porous polyurethane. Our findings can lead to the development of novel x-ray shielding products that can reduce the physical and mental stress on users.

[Pulmonary Artery Transection in Surgery for Coronary Artery Fistula].

Coronary artery fistula is a rare abnormality in the communication between a coronary artery and any of the cardiac chambers or major vessels. At present, there is no standard surgical treatment and the most appropriate method is selected on a case-by-case basis. We report one case of coronary artery fistulae in which pulmonary artery transection was required around the left main trunk (LMT). A 62-year-old man who had coronary artery fistulae with an aneurysm which increased from 12 mm to 16 mm in a two-year span. The fistula was located adjacent to the LMT. A complete aneurysm excision under cardiopulmonary bypass was performed, which required pulmonary artery transection. No postoperative complications occurred. Postoperative coronary computed tomography scan showed intact coronary arteries and complete aneurysm removal.

Overexpression of polyphosphate polymerases and deletion of polyphosphate phosphatases shorten the replicative lifespan in yeast.

We previously found that overexpression of phosphate starvation-responsive genes by disrupting PHO80 led to a shortened replicative lifespan in yeast. To identify lifespan-related genes, we screened upregulated genes in the pho80Δ mutant and focused on the VTC genes, which encode the vacuolar polyphosphate (polyP) polymerase complex. VTC1/VTC2/VTC4 deletion restored the lifespan and intracellular polyP levels in pho80Δ. In the wild type, overexpression of VTC5 or a combination of the other VTCs caused high polyP accumulation and shortened lifespan. Similar phenotypes were caused by the deletion of polyP phosphatase genes-vacuolar PPN1 and cytosolic PPX1. The polyP-accumulating strains exhibited stress sensitivities. Thus, we demonstrated that polyP metabolic enzymes participate in replicative lifespan, and extreme polyP accumulation shortens the lifespan.

Searching for Osmosensing Determinants in Poplar Histidine-Aspartate Kinases.

Previous works have shown the existence of protein partnership, belonging to a MultiStep Phosphorelay (MSP), potentially involved in osmosensing in Populus. The first actor of this signalling pathway belongs to the histidine-aspartate kinase (HK) family, which also includes the yeast osmosensor Sln1, as well as the Arabidopsis putative osmosensor AHK1. In poplar, the homologous AHK1 protein corresponds to a pair of paralogous proteins, HK1a and HK1b, exhibiting an extracellular domain (ECD), as in Sln1 and AHK1. An ECD alignment of AHK1-like proteins, from different plant species, showed a particularly well conserved ECD and revealed the presence of a cache domain. This level of conservation suggested a functional role of this domain in osmosensing. Thus, we tested this possibility by modelling assisted mutational analysis of the cache domain of the Populus HK1 proteins. The mutants were assessed for their ability to respond to different osmotic stress and the results point to an involvement of this domain in HK1 functionality. Furthermore, since HK1b was shown to respond better to stress than HK1a, these two receptors constituted a good system to search for osmosensing determinants responsible for this difference in efficiency. With domain swapping experiments, we finally demonstrated that the cache domain, as well as the second transmembrane domain, are involved in the osmosensing efficiency of these receptors.

The pH-sensing Rim101 pathway regulates cell size in budding yeast.

Although cell size regulation is crucial for cellular functions in a variety of organisms from bacteria to humans, the underlying mechanisms remain elusive. Here, we identify Rim21, a component of the pH-sensing Rim101 pathway, as a positive regulator of cell size through a flow cytometry-based genome-wide screen of Saccharomyces cerevisiae deletion mutants. We found that mutants defective in the Rim101 pathway were consistently smaller than wildtype cells in the log and stationary phases. We show that the expression of the active form of Rim101 increased the size of wildtype cells. Furthermore, the size of wildtype cells increased in response to external alkalization. Microscopic observation revealed that this cell size increase was associated with changes in both vacuolar and cytoplasmic volume. We also found that these volume changes were dependent on Rim21 and Rim101. In addition, a mutant lacking Vph1, a component of V-ATPase that is transcriptionally regulated by Rim101, was also smaller than wildtype cells, with no increase in size in response to alkalization. We demonstrate that the loss of Vph1 suppressed the Rim101-induced increase in cell size under physiological pH conditions. Taken together, our results suggest that the cell size of budding yeast is regulated by the Rim101-V-ATPase axis under physiological conditions as well as in response to alkaline stresses.

Roles of phosphatidylserine and phospholipase C in the activation of TOR complex 2 signaling in Saccharomyces cerevisiae.

Target of rapamycin (TOR) forms two distinct complexes, TORC1 and TORC2, to exert its essential functions in cellular growth and homeostasis. TORC1 signaling is regulated in response to nutrients such as amino acids and glucose; however, the mechanisms underlying the activation of TORC2 signaling are still poorly understood compared to those for TORC1 signaling. In the budding yeast Saccharomyces cerevisiae, TORC2 targets the protein kinases Ypk1 and Ypk2 (hereafter Ypk1/2), and Pkc1 for phosphorylation. Plasma membrane stress is known to activate TORC2-Ypk1/2 signaling. We have previously reported that methylglyoxal (MG), a metabolite derived from glycolysis, activates TORC2-Pkc1 signaling. In this study, we found that MG activates the TORC2-Ypk1/2 and TORC2-Pkc1 signaling, and that phosphatidylserine is involved in the activation of both signaling pathways. We also demonstrated that the Rho family GTPase Cdc42 contributes to the plasma membrane stress-induced activation of TORC2-Ypk1/2 signaling. Furthermore, we revealed that phosphatidylinositol-specific phospholipase C, Plc1, contributes to the activation of both TORC2-Ypk1/2 and TORC2-Pkc1 signaling.

Impact of the COVID-19 Pandemic on Prehospital Intervention and Survival of Patients With Out-of-Hospital Cardiac Arrest in Osaka City, Japan.

BACKGROUND: The coronavirus disease (COVID-19) pandemic may have influenced the prehospital emergency care and deaths of individuals experiencing an out-of-hospital cardiac arrest (OHCA).Methods and Results: We analyzed the registry data of 2,420 and 2,371 OHCA patients in Osaka City, Japan in 2019 and 2020, respectively, according to the 3 waves of the COVID-19 pandemic. Patient outcomes were compared using multivariable logistic regression analyses with the 2019 data as the reference. Bystander cardiopulmonary resuscitation (CPR) was initiated significantly less frequently in 2020 than in 2019 (2019: 48.0%, 2020: 42.7%, P<0.001), particularly during the first wave (2019: 47.2%, 2020: 42.9%, P=0.046) and second wave (2019: 48.1%, 2020: 41.2%, P=0.010), but not during the third wave (2019: 49.2%, 2020: 44.1%, P=0.066). The public-access automated external defibrillator was less frequently applied during the first wave (2019: 12.6%, 2020: 9.9%, P=0.043), with no significant difference during the second wave (2019: 12.5%, 2020: 12.8%, P=0.863) and third wave (2019: 13.7%, 2020: 13.0%, P=0.722). There was a significant difference in 1-month survival with favorable neurological outcomes (2019: 4.6%, 2020: 3.3%, P=0.018), with a 28% reduction in the adjusted odds ratio in 2020 (0.72; 95% confidence interval: 0.52-0.99, P=0.044). CONCLUSIONS: Bystander CPR and neurologically favorable outcomes after OHCA decreased significantly during the COVID-19 pandemic in Japan.

The oncogene-dependent resistance to reprogramming unveils cancer therapeutic targets.

The resistance to transcription factor-mediated reprogramming into pluripotent stem cells is one of the distinctive features of cancer cells. Here we dissect the profiles of reprogramming factor binding and the subsequent transcriptional response in cancer cells to reveal its underlying mechanisms. Using clear cell sarcomas (CCSs), we show that the driver oncogene EWS/ATF1 misdirects the reprogramming factors to cancer-specific enhancers and thereby impairs the transcriptional response toward pluripotency that is otherwise provoked. Sensitization to the reprogramming cue is observed in other cancer types when the corresponding oncogenic signals are pharmacologically inhibited. Exploiting this oncogene dependence of the transcriptional "stiffness," we identify mTOR signaling pathways downstream of EWS/ATF1 and discover that inhibiting mTOR activity substantially attenuates the propagation of CCS cells in vitro and in vivo. Our results demonstrate that the early transcriptional response to cell fate perturbations can be a faithful readout to identify effective therapeutics targets in cancer cells.

Coordinated regulation of TORC2 signaling by MCC/eisosome-associated proteins, Pil1 and tetraspan membrane proteins during the stress response.

MCCs are linear invaginations of the yeast plasma membrane that form stable membrane microdomains. Although over 20 proteins are localized in the MCCs, it is not well understood how these proteins coordinately maintain normal MCC function. Pil1 is a core eisosome protein and is responsible for MCC-invaginated structures. In addition, six-tetraspan membrane proteins (6-Tsp) are localized in the MCCs and classified into two families, the Sur7 family and Nce102 family. To understand the coordinated function of these MCC proteins, single and multiple deletion mutants of Pil1 and 6-Tsp were generated and their MCC structure and growth under various stresses were investigated. Genetic interaction analysis revealed that the Sur7 family and Nce102 function in stress tolerance and normal eisosome assembly, respectively, by cooperating with Pil1. To further understand the role of MCCs/eisosomes in stress tolerance, we screened for suppressor mutants using the SDS-sensitive phenotype of pil1Δ 6-tspΔ cells. This revealed that SDS sensitivity is caused by hyperactivation of Tor kinase complex 2 (TORC2)-Ypk1 signaling. Interestingly, inhibition of sphingolipid metabolism, a well-known downstream pathway of TORC2-Ypk1 signaling, did not rescue the SDS-sensitivity of pil1Δ 6-tspΔ cells. These results suggest that Pil1 and 6-Tsp cooperatively regulate TORC2 signaling during the stress response.

Regulation of sphingolipid biosynthesis in the endoplasmic reticulum via signals from the plasma membrane in budding yeast.

Saccharomyces cerevisiae LIP1 encodes a regulatory subunit that forms a complex with the ceramide synthase catalytic subunits, Lag1/Lac1, which is localized on the membrane of endoplasmic reticulum. To understand the underlying regulatory mechanism of sphingolipid biosynthesis, we generated strains upon replacing the chromosomal LIP1 promoter with a Tet-off promoter, which enables the expression in Dox-dependent manner. The lip1-1 strain, obtained through the promoter substitution, exhibits severe growth inhibition and remarkable decrease in sphingolipid synthesis in the presence of Dox. Using this strain, we investigated the effect of a decrease in ceramide synthesis on TOR complex 2 (TORC2)-Ypk1 signaling, which senses the complex sphingolipid level at the plasma membrane and promotes sphingolipid biosynthesis. In lip1-1 cells, Ypk1 was activated via both upstream kinases, TORC2 and yeast PDK1 homologues, Pkh1/2, thereby inducing hyperphosphorylation of Lag1, but not of another Ypk1-substrate, Orm1, which is a known negative regulator of the first step of sphingolipid metabolism, in the presence of Dox. Therefore, our data suggest that the metabolic enzyme activities at each step of the sphingolipid biosynthetic pathway are controlled through a fine regulatory mechanism.

High prevalence of Mycoplasma equirhinis in Thoroughbred horses with respiratory symptoms in autumn 2018.

Mycoplasma species are often isolated from horses with respiratory symptoms; however, the pathogenicity of Mycoplasma is still unclear. In autumn of 2018, we encountered an increase in cases with respiratory symptoms, mainly coughing, in a group of Thoroughbred racehorses in Japan. We examined tracheal wash samples obtained from 40 of those cases. Bacteria and viruses that commonly cause respiratory symptoms were investigated, and anaerobes were detected in only 5 cases and Streptococcus equi subsp. zooepidemicus (S. zooepidemicus) was detected in only 1 case of 40 cases with loop-mediated isothermal amplification assay. S. zooepidemicus and Streptococcus pneumoniae were isolated at a bacterial count of higher than 1.0 × 104 CFU/ml from 5 and 2 cases of 28 cases cultured, respectively. None of the viruses investigated was detected in 40 cases. Mycoplasma equirhinis (M. equirhinis) was isolated from 40.0% (16/40) of the cases, which was higher than previously reported isolation rates. The rate of M. equirhinis isolation in the cases from 2018 was significantly higher than the isolation rates in the other horses: clinical cases with respiratory symptoms in 2019-2020 (13.6%, 3/22) and healthy horses (13.5%, 5/37) in Japan. In this study, the isolation rate of M. equirhinis from horse group with cough symptoms in 2018 was high and no other common etiological agents were detected. The pathogenesis of M. equirhinis is still unclear, however, M. equirhinis might have been associated with respiratory symptoms in the Thoroughbred horse cases in 2018.

A glutamine sensor that directly activates TORC1.

TOR complex 1 (TORC1) is an evolutionarily-conserved protein kinase that controls cell growth and metabolism in response to nutrients, particularly amino acids. In mammals, several amino acid sensors have been identified that converge on the multi-layered machinery regulating Rag GTPases to trigger TORC1 activation; however, these sensors are not conserved in many other organisms including yeast. Previously, we reported that glutamine activates yeast TORC1 via a Gtr (Rag ortholog)-independent mechanism involving the vacuolar protein Pib2, although the identity of the supposed glutamine sensor and the exact TORC1 activation mechanism remain unclear. In this study, we successfully reconstituted glutamine-responsive TORC1 activation in vitro using only purified Pib2 and TORC1. In addition, we found that glutamine specifically induced a change in the folding state of Pib2. These findings indicate that Pib2 is a glutamine sensor that directly activates TORC1, providing a new model for the metabolic control of cells.

A novel algorithm for extracting soft-tissue and bone images measured using a photon-counting type X-ray imaging detector with the help of effective atomic number analysis.

Most of the objects targeted for X-ray examination are composed of soft-tissue and bone. We aimed to develop an algorithm for generating X-ray images which can give quantitative information of soft-tissue and bone using an energy-resolving photon-counting type imaging detector. We used polychromatic X-rays for analysis in which both the beam hardening effect and detector response were properly corrected and then succeeded in virtually treating the amount of measured X-ray attenuation as if it were measured using monochromatic X-rays.

A disposable OSL dosimeter for in vivo measurement of rectum dose during brachytherapy.

PURPOSE: We aimed to develop a disposable rectum dosimeter and to demonstrate its ability to measure exposure dose to the rectum during brachytherapy for cervical cancer treatment using high-dose rate 192 Ir. Our rectum dosimeter measures the dose with an optically stimulated luminescence (OSL) sheet which was furled to a catheter. The catheter we used is 6 mm in diameter; therefore, it is much less invasive than other rectum dosimeters. The rectum dosimeter developed in this study has the characteristics of being inexpensive and disposable. It is also an easy-to-use detector that can be individually sterilized, making it suitable for clinical use. METHODS: To obtain a dose calibration curve, phantom experiments were performed. Irradiation was performed using a cubical acrylic phantom, and the response of the OSL dosimeter was calibrated with the calculation value predicted by the treatment planning system (TPS). Additionally, the dependence of catheter angle on the dosimeter position and repeatability were evaluated. We also measured the absorbed dose to the rectum of patients who were undergoing brachytherapy for cervical cancer (n = 64). The doses measured with our dosimeters were compared with the doses calculated by the TPS. In order to examine the causes of large differences between measured and planned doses, we classified the data into common and specific cases when performing this clinical study. For specific cases, the following three categories were considered: (a) patient movement, (b) gas in the vagina and/or rectum, and (c) artifacts in the X-ray image caused by applicators. RESULTS: A dose calibration curve was obtained in the range of 0.1 Gy-10.0 Gy. From the evaluation of the dependence of catheter angle on the dosimeter position and repeatability, we determined that our dosimeter can measure rectum dose with an accuracy of 3.1% (k = 1). In this clinical study, we succeeded in measuring actual doses using our rectum dosimeter. We found that the deviation of the measured dose from the planned dose was derived to be 12.7% (k = 1); this result shows that the clinical study included large elements of uncertainty. The discrepancies were found to be due to patient motion during treatment, applicator movement after planning images were taken, and artifacts in the planning images. CONCLUSIONS: We present the idea that a minimally invasive rectum dosimeter can be fabricated using an OSL sheet. Our clinical study demonstrates that a rectum dosimeter made from an OSL sheet has sufficient ability to evaluate rectum dose. Using this dosimeter, valuable information concerning organs at risk can be obtained during brachytherapy.

Effective atomic number image determination with an energy-resolving photon-counting detector using polychromatic X-ray attenuation by correcting for the beam hardening effect and detector response.

In this study, we propose an effective atomic number (Zeff) determination method based on a photon-counting technique. The proposed method can correct for the beam hardening effect and detector response based on polychromatic X-rays to allow high accuracy material identification. To demonstrate the effectiveness of our method, the procedure was applied to X-ray images acquired by a prototype energy-resolving photon-counting detector and we obtained an Zeff image with accuracy of Zeff ± 0.5 regardless of the mass thickness.

Yeast-based reporter assay system for identifying the requirements of intramembrane proteolysis by signal peptide peptidase of Arabidopsis thaliana.

Signal peptide peptidase (SPP) is an aspartic protease with two active sites, YD and GXGD, in the transmembrane domain. SPP cleaves signal peptides, and the released fragments play key roles in the immune system, embryo development and protein turnover in cells. Despite SPP having an important function, a general system to identify the requirements of intramembrane proteolysis by SPP has not been developed because proteolysis occurs in the membrane. In this study, we first established a reporter assay system in yeast to verify the cleavage activity of the Arabidopsis thaliana SPP (AtSPP). Next, we screened candidate substrates of AtSPP from A. thaliana pollen and roots. In the pollen, 13 signal peptides with 'pollen' and 'cell wall' as gene ontology terms were selected. In the roots, mutants overexpressing AtSPP were constructed, and gene expression changes were compared with the wild-type. Nine signal peptides expressed in the roots were selected. Then we used the candidate substrates in our reporter assay system to determine the requirements for proteolysis by AtSPP. Fifteen of 22 signal peptides were cleaved by AtSPP. The absence of the positively charged amino acids, His and Lys on the C terminus of the signal sequence, was observed in cleaved substrates. Moreover, mutation of a helix breaker-to-Leu substitution in the intramembrane region in substrates prevented cleavage by AtSPP. These results indicated that substrates of AtSPP required the helix breaker structure to be cleaved.

New Insight into HPts as Hubs in Poplar Cytokinin and Osmosensing Multistep Phosphorelays: Cytokinin Pathway Uses Specific HPts.

We have previously identified proteins in poplar which belong to an osmosensing (OS) signaling pathway, called a multistep phosphorelay (MSP). The MSP comprises histidine-aspartate kinases (HK), which act as membrane receptors; histidine phosphotransfer (HPt) proteins, which act as phosphorelay proteins; and response regulators (RR), some of which act as transcription factors. In this study, we identified the HK proteins homologous to the Arabidopsis cytokinin (CK) receptors, which are first partners in the poplar cytokinin MSP, and focused on specificity of these two MSPs (CK and OS), which seem to share the same pool of HPt proteins. Firstly, we isolated five CK HKs from poplar which are homologous to Arabidopsis AHK2, AHK3, and AHK4, namely, HK2, HK3a, HK3b, HK4a, HK4b. These HKs were shown to be functional kinases, as observed in a functional complementation of a yeast HK deleted strain. Moreover, one of these HKs, HK4a, was shown to have kinase activity dependent on the presence of CK. Exhaustive interaction tests between these five CK HKs and the 10 HPts characterized in poplar were performed using two-hybrid and BiFC experiments. The resulting partnership was compared to that previously identified between putative osmosensors HK1a/1b and HPt proteins. Finally, in planta coexpression analysis of genes encoding these potential partners revealed that almost all HPts are coexpressed with CK HKs in four different poplar organs. Overall, these results allowed us to unravel the common and specific partnerships existing between OS and CK MSP in Populus.