Development of fast neutron pinhole camera using nuclear emulsion for neutron emission profile measurement in KSTAR.

We have developed a compact fast neutron camera based on a stack of nuclear emulsion plates and a pinhole collimator. The camera was installed at J-port of Korea superconducting tokamak advanced research at National Fusion Research Institute, Republic of Korea. Fast neutron images agreed better with calculated ones based on Monte Carlo neutron simulation using the uniform distribution of Deuterium-Deuterium (DD) neutron source in a torus of 40 cm radius.

Progress in development of neutron energy spectrometer for deuterium plasma operation in KSTAR.

Two types of DD neutron energy spectrometer (NES) are under development for deuterium plasma operation in KSTAR to understand behavior of beam ions in the plasma. One is based on the state-of-the-art nuclear emulsion technique. The other is based on a coincidence detection of a recoiled proton and a scattered neutron caused by an elastic scattering of an incident DD neutron, which is called an associated particle coincidence counting-NES. The prototype NES systems were installed at J-port in KSTAR in 2012. During the 2012 and 2013 experimental campaigns, multiple shots-integrated neutron spectra were preliminarily obtained by the nuclear emulsion-based NES system.

SU-E-T-234: LET Measurement Using Nuclear Emulsion and Monte Carlo Simulation for Proton Beam.

PURPOSE: The significant issue of particle therapy such as proton and carbon ion biological effect on tumors and normal tissue. This effect closely connected with linear-energy-transfer (LET). This work presents a Monte Carlo study using GEANT4 and the verification using Nuclear Emulsion to show LET for proton beam. METHODS: National Cancer Center (NCC) has IBA Beam Nozzle and cyclotron for proton therapy. We use proton beam bragg peak range 14cm. Also, we already developed the simulation using GEANT4 and finished validation for scattering proton beam. In our simulation, we make same condition with experimental setup.Nuclear emulsion films interleaved with tissue equivalent absorbers can be fruitfully used to reconstruct proton tracks with very high precision. This Nuclear emulsion film has been supported from Nagoya University, analyzed in Pusan University, was irradiated with a therapeutic proton beam at NCC. The Emulsion packs was located at entrance and bragg peak region of proton. This position means low and high LET region. The scanning of the emulsions has been performed at Nagoya University, where a fully automated microscopic scanning technology has been developed for the OPERA experiment on neutrino oscillations. RESULTS: We could see the reconstructed track of proton scanning emulsion. From film scanning, we got the LET distribution at low and high LET region for several proton tracks. Simulation results was similar distribution within standard deviation in acceptance level. Also we got the RBE distribution using LET measurement for proton beam. CONCLUSIONS: We measured LET at entrance and bragg peak region using Monte Carlo study and Nuclear Emulsion film, for NCC proton beam. This results means the good observation of LET using the nuclear emulsion. And this method can be used successfully in medical field.

Bio rapid prototyping by extruding/aspirating/refilling thermoreversible hydrogel.

This paper reports a method for rapid prototyping of cell tissues, which is based on a system that extrudes, aspirates and refills a mixture of cells and thermoreversible hydrogel as a scaffold. In the extruding mode, a cell-mixed scaffold solution in the sol state is extruded from a cooled micronozzle into a temperature-controlled substrate, which keeps the scaffold in the gel state. In the aspiration mode, the opposite process is performed by Bernoulli suction. In the refilling mode, the solution is extruded into a groove created in the aspiration mode. The minimum width of extruded hydrogel pattern is 114 +/- 15 microm by employing a nozzle of diameter 100 microm, and that of aspirated groove was 355 +/- 10 microm using a 500 microm-diameter nozzle. Gum arabic is mixed with the scaffold solution to avoid peeling-off of the gel pattern from the substrate. Patterning of Sf-9 cell tissue is demonstrated, and the stability of the patterned cell is investigated. This system offers a procedure for rapid prototyping and local modification of cell scaffolds for tissue engineering.

Living donor liver transplantation for congenital absence of the portal vein.

The congenital absence of the portal vein (CAPV) is a rare venous malformation in which mesenteric venous blood drains directly into the systemic circulation. Liver transplantation (OLT) may be indicated for patients with symptomatic CAPV refractory to medical treatment, especially due to hyperammonemia, portosystemic encephalopathy, hepatopulmonary syndrome, or hepatic tumors. Because portal hypertension and collateral circulation do not occur with CAPV, significant splanchnic congestion may occur when the portocaval shunt is totally clamped during portal vein (PV) reconstruction in OLT. This phenomenon results in severe bowel edema and hemodynamic instability, which negatively impact the patient's condition and postoperative recovery. We have successfully reconstructed the PV in living donor liver transplantation (LDLT) using a venous interposition graft, which was anastomosed end-to-side to the portocaval shunt by a partial side-clamp, using a patent round ligament of the liver, which was anastomosed end-to-end to the graft PV with preservation of both the portal and caval blood flows. Owing to the differences in anatomy among patients, at LDLT for CAPV liver transplant surgeons should seek to preserve both portal and caval blood flows.

Formation of globules and aggregates of DNA chains in DNA/polyethylene glycol/monovalent salt aqueous solutions.

It has been known that giant DNA shows structural transitions in aqueous solutions under the existence of counterions and other polymers. However, the mechanism of these transitions has not been fully understood. In this study, we directly observed structures of probed (dye-labeled), dilute DNA chains in unprobed DNA/polyethylene glycol (PEG)/monovalent salt (NaCl) aqueous solutions with fluorescent microscopy to examine this mechanism. Specifically, we varied the PEG molecular weight and salt concentration to investigate the effect of competition between the depletion and electrostatic interactions on the coil-globule transition and the aggregate formation. It was found that the globules coexist with the aggregates when the unprobed DNA chains have a concentration higher than their overlap concentration. We discuss the stability of the observed structures on the basis of a free energy model incorporating the attractive depletion energy, the repulsive electrostatic energy, and the chain bending energy. This model suggested that both of the globules and aggregates are more stable than the random coil at high salt concentrations/under existence of PEG and the transition occurs when the depletion interaction overwhelms the electrostatic interaction. However, the coexistence of the globule and aggregate was not deduced from the thermodynamic model, suggesting a nonequilibrium aspect of the DNA solution and metastabilities of these structures. Thus, the population ratio of globules and aggregates was also analyzed on the basis of a kinetic model. The analysis suggested that the depletion interaction dominates this ratio, rationalizing the coexistence of globules and aggregates.

[Ross procedure with aortic replacement for aortic stenosis with ascending aortic aneurysm: report of a case].

Bicuspid aortic valve (BAV) is a common congenital heart disease, and it is well known to be a risk factor for ascending aortic dilatation and dissection. We here report a case of 34-year-old woman who underwent Ross procedure with ascending aortic replacement under the diagnosis of subaortic stenosis and ascending aortic aneurysm. She was pointed out to have heart murmur soon after the birth diagnosed as patent ductus arteriosus. The ductus was ligated when she was 3-years-old, however, heart murmur remained. Further examinations revealed that she also had aortic stenosis with BAV. During her 20-year-follow-up, subaortic stenosis and ascending aorta ectasia were also progressed. Pathological examinations of resected ascending aortic wall showed mucoid degeneration and laceration of collagen fibers, suggesting the fragility of dilated aortic wall with BAV.

Meiotic hybridogenesis in triploid Misgurnus loach derived from a clonal lineage.

Triploid loaches Misgurnus anguillicaudatus are derived from unreduced diploid gametes produced by an asexual clonal lineage that normally undergoes gynogenetic reproduction. Here, we have investigated the reproductive system of two types of triploids: the first type carried maternally inherited clonal diploid genomes and a paternally inherited haploid genome from the same population; the second type had the same clonal diploid genomes but a haploid genome from another, genetically divergent population. The germinal vesicles of oocytes from triploid females (3n=75) contained only 25 bivalents, that is, 50 chromosomes. Flow cytometry revealed that the majority of the progeny resulting from fertilization of eggs from triploid females with normal haploid sperm were diploid. This indicates that triploid females mainly produced haploid eggs. Microsatellite analyses of the diploid progeny of triploid females showed that one allele of the clonal genotype was not transmitted to haploid eggs. Moreover, the identity of the eliminated allele differed between the two types of triploids. Our results demonstrate that there is preferential pairing of homologous chromosomes as well as the elimination of unmatched chromosomes in the course of haploid egg formation, that is, meiotic hybridogenesis. Two distinct genomes in the clone suggest its hybrid origin.

Centrosome amplification induced by survivin suppression enhances both chromosome instability and radiosensitivity in glioma cells.

Glioblastoma is characterised by invasive growth and a high degree of radioresistance. Survivin, a regulator of chromosome segregation, is highly expressed and known to induce radioresistance in human gliomas. In this study, we examined the effect of survivin suppression on radiosensitivity in malignant glioma cells, while focusing on centrosome aberration and chromosome instability (CIN). We suppressed survivin by small interfering RNA transfection, and examined the radiosensitivity using a clonogenic assay and a trypan blue exclusion assay in U251MG (p53 mutant) and D54MG (p53 wild type) cells. To assess the CIN status, we determined the number of centrosomes using an immunofluorescence analysis, and the centromeric copy number by fluorescence in situ hybridisation. As a result, the radiosensitisation differed regarding the p53 status as U251MG cells quickly developed extreme centrosome amplification (=CIN) and enhanced the radiosensitivity, while centrosome amplification and radiosensitivity increased more gradually in D54MG cells. TUNEL assay showed that survivin inhibition did not lead to apoptosis after irradiation. This cell death was accompanied by an increased degree of aneuploidy, suggesting mitotic cell death. Therefore, survivin inhibition may be an attractive therapeutic target to overcome the radioresistance while, in addition, proper attention to CIN (centrosome number) is considered important for improving radiosensitivity in human glioma.

The forkhead-associated domain of NBS1 is essential for nuclear foci formation after irradiation but not essential for hRAD50[middle dot]hMRE11[middle dot]NBS1 complex DNA repair activity.

NBS1 (p95), the protein responsible for Nijmegen breakage syndrome, shows a weak homology to the yeast Xrs2 protein at the N terminus region, known as the forkhead-associated (FHA) domain and the BRCA1 C terminus domain. The protein interacts with hMRE11 to form a complex with a nuclease activity for initiation of both nonhomologous end joining and homologous recombination. Here, we show in vivo direct evidence that NBS1 recruits the hMRE11 nuclease complex into the cell nucleus and leads to the formation of foci by utilizing different functions from several domains. The amino acid sequence at 665-693 on the C terminus of NBS1, where a novel identical sequence with yeast Xrs2 protein was found, is essential for hMRE11 binding. The hMRE11-binding region is necessary for both nuclear localization of the complex and for cellular radiation resistance. On the other hand, the FHA domain regulates nuclear foci formation of the multiprotein complex in response to DNA damage but is not essential for nuclear transportation of the complex and radiation resistance. Because the FHA/BRCA1 C terminus domain is widely conserved in eukaryotic nuclear proteins related to the cell cycle, gene regulation, and DNA repair, the foci formation could be associated with many phenotypes of Nijmegen breakage syndrome other than radiation sensitivity.

Microsatellite-centromere mapping in the loach, Misgurnus anguillicaudatus.

Primer sets for 15 polymorphic microsatellite loci were developed in the loach, Misgurnus anguillicaudatus (Cobitidae) by molecular cloning and sequencing techniques. Mendelian inheritance was confirmed for the 15 loci by examining the genotypic segregation produced with the primer sets in two full-sib families. The loci were mapped in relation to their centromere in four gynogenetic diploid lines, which were induced by inhibition of the second meiotic division after fertilization with genetically inert sperm. Microsatellite-centromere recombination rates ranged between 0.06 and 0.95 under the assumption of complete interference. Thus, these loci are distributed from the centromeres to the telomeres of their respective chromosomes. The success of mitotic gynogenesis, produced by suppression of the first cleavage, was verified by homozygosity at three diagnostic microsatellite loci that exhibited high gene-centromere meiotic recombination rates in the same family. The differences in heterozygosity levels observed with these markers were attributed to differences in the temporal application of heat shock following inert sperm activation.

Expression of full-length NBS1 protein restores normal radiation responses in cells from Nijmegen breakage syndrome patients.

Cells from Nijmegen breakage syndrome (NBS) display multiple phenotypes, such as chromosomal instability, hypersensitivity to cell killing from ionizing radiation, and possibly abnormal cell cycle checkpoints. NBS1, a gene mutated in NBS patients, appears to encode a possible repair protein, which could form the foci of a sensor-like molecular complex capable of detecting DNA double strand breaks, however, it has no kinase domain for signaling DNA damage. Here, we report that the stable expression of NBS1 cDNA in NBS cells after transfection results in the complete restoration of foci formation in the nucleus, and in normal cell survival after irradiation. The prolonged G2 block observed after irradiation was also abolished by expression of NBS1, providing additional confirmation that the G2 checkpoint is abrogated in NBS cells. These results suggest that a defective NBS1 protein could be the sole cause of the NBS phenotype, and that NBS1 likely interacts with another protein(s) to produce the entire range of NBS phenotypic expression.

A polymorphic CA repeat marker at the human 27-kD calbindin (CALB1) locus.

A polymorphic dinucleotide (CA) sequence was isolated from a BAC clone containing the human 27-kD calbindin (CALB1) gene at 8q21. This polymorphism will be a useful genetic marker to study genetic variations of the CALB1 gene.

Banding of the pulmonary trunk in preparation for a Fontan operation.

Banding of the pulmonary trunk is an important surgical procedure for patients who have congenital cardiac malformations with unrestricted pulmonary flow. We propose a new concept for determining in such circumstances the most appropriate length of the band used to constrict the pulmonary trunk in preparation for a Fontan operation. We studied 14 patients undergoing banding of the pulmonary trunk and measured the following parameters: diameter of aorta, diameter of pulmonary trunk, length of pulmonary arterial band and maximum flow velocity across the banded segment. We calculated an index from our original parameter based on the formula; length of band/(diameter of aorta diameter of pulmonary trunk). The diameter of aorta was 9.5 +/- 1.4 mm, and that of the pulmonary trunk was 9.6 +/- 2.3 mm. The length of the band was 16.5 +/- 3.4 mm, giving a calculated index of 0.188 +/- 0.038. The maximum flow velocity was 4.02 +/- 0.46 m/s. No correlation was found between the length of the band and body weight, and also no correlation was found between the length of the band and maximum flow velocity. The calculated index had a negative correlation with the maximum velocity of flow across the band (y = -8.13x +/- 5.56, R = 0.74, p < 0.01). We believe that the proposed index is a useful guide in determining the length of a pulmonary band when preparing patients for a Fontan operation.

[A case of severe cytomegalovirus infection after the repair of coarctation of aorta].

We report a 2-month-old boy without any immuno-compromised diseases, who suffered from the severe cytomegalovirus (CMV) infection after the subclavian flap aortoplasty and pulmonary artery banding for coarctation complex. He underwent the operation at 2 months old and received 2 units of irradiated packed red blood cells before and after the surgery. His postoperative course was uneventful but the interstitial pneumonitis, until he developed watery diarrhea 10 days after the surgery following hepatitis with the marked hepatomegaly 3 weeks after. Since CMV infection was confirmed as the cause of the pneumonitis, enterocolitis and hepatitis, he was initially treated by gamma-globulin with the high CMV titer at a dose of 200 mg/kg/day for 2 days and ganciclovir at a dose of 10 mg/kg/day for 14 days. Because of the persistent CMV infection, he needed two more treatments of ganciclovir at the same dosage and gamma-globulin once a week for 2 months. He finally recovered from severe CMV infection 5 months after the above treatments. In conclusion, the severe CMV infection can occur by blood transfusion even in the surgical case with normal immune system. If one finds pneumonitis, hepatitis or enterocolitis after any type of surgery with history of blood transfusion, CMV infection should be suspected as the cause of these diseases.

[One-stage repair of interrupted aortic arch with selective cerebral perfusion].

To minimize the neurological compromise after the circulatory arrest, the selective cerebral perfusion could be beneficial. We underwent one-stage repair of the interrupted aortic arch (IAA) with various intracardiac anomalies for the six patients, age ranging from 12 days to 4 months, by using the selective cerebral perfusion. Cardiopulmonary bypass was established by using two-way arterial cannulation supported by the two respective pump systems, one of which utilized the EPTFE graft anastmosed to either the bracheocephalic artery or the right subclavian artery and second of which enrouted through the arterial ductus to the descending aorta. The cerebral perfusion during the circulatory arrest for the aortic arch repair was maintained by the selective perfusion via EPTFE graft with 10 ml/kg/min blood flow. After the completion of the arch repair, the total system perfusion was restarted through the graft and the repair of the intracardiac anomalies was followed. Of six, no operative death or neurological complications related to the operation were found. The clinical neurological evaluation after operation also demonstrated the normal for the age. In conclusion, the selective cerebral perfusion by using the EPTFE graft during the circulatory arrest might decrease the risk of brain damage.

[Successful repair of a newborn case with total anomalous pulmonary venous connection presenting hemodynamically common pulmonary vein atresia].

Common pulmonary vein atresia is a very rare from of congenital cardiac anomaly in which the common chamber of pulmonary veins has no direct communication to the heart or systemic venous system. We report here on one-day-old boy with total anomalous pulmonary venous connection having severe stenosis (sized less than one millimeter in diameter) of vertical vein between common pulmonary vein and superior vena cava. His anatomic feature was considered as a spectrum between common pulmonary vein atresia and total anomalous pulmonary venous connection because of almost atretic draining vertical vein. His clinical presentation and hemodynamic characteristics were the same as common pulmonary vein atresia. He underwent an emergent anastomosis of common pulmonary vein to left atrium at 33 hours after birth under cardiopulmonary bypass, cross-sectional echocardiography and Doppler color mapping. He was discharged home two months after the surgery. Cardiac catheterization after six months revealed normal cardiac function without any evidence of pulmonary venous chamber stenosis. To improve the outcome of surgical intervention for this life-threatening cardiac anomaly, immediate the precise diagnosis using echocardiography followed by emergent surgical correction is in dispensable.

Gender differences in effects of 20 days horizontal bed rest on muscle strength in young subjects.

Gender differences in the effect of 20 days bed rest (BR) on muscle strength were evaluated in voluntary 11 male and 7 female students. Maximum Isometric Voluntary Contractions (MVC) of 4 right arm muscles (RAM), 5 right leg muscles (RLM), and 2 body trunk muscles were measured with an isometric dynamometer, respectively. Muscle masses (MM) of right arm and leg and body trunk were determined by dual energy X-ray absorptiometry, respectively. The maximum cross sectional area (CSAmax) of right m. quadriceps femoris was measured by magnetic resonance imaging. Elbow flexion MVC in males and all MVC of RLM except knee flexion in both males and females were decreased (p<0.05), but elbow extension MVC in females was increased (p<0.05), while all of other MVC only tended to decrease. However, the decrements in leg MVC were not correlated to the leg MM, and also the decrement in knee extension was not correlated to the CSAmax of m. quadriceps. The reduction of MVC of antigravity muscles might be caused not only by a decrease in MM but also by other factors. The greater decrements of leg MVC during BR were the higher initial level in males, but the inverse was observed in females. However, this discrepancy between males and females cannot be explained in the present study.