Search

Necrotizing enterocolitis (NEC), focal intestinal perforation (FIP), and meconium-related ileus (MRI) are major diseases that cause gastrointestinal disorders in extremely low-birth-weight infants (ELBWIs). We conducted a review to compare the postoperative outcomes of ELBWIs with these diseases in our neonatal intensive-care unit. A retrospective chart review of ELBWIs surgically treated for NEC (n = 31), FIP (n = 35), and MRI (n = 16) in 2001–2018 was undertaken. This period was divided into early (2001–2005), middle (2006–2010), and late (2011–2018) periods. Data were analyzed with the Cochran-Armitage test. Statistical significance was defined as p

During the cylindrical traverse grinding of a slender workpiece, the ground workpiece is easily bent by the normal grinding force owing to its low stiffness. Therefore, it is difficult to finish the slender workpiece with high accuracy. To prevent the elastic deformation of a workpiece during the grinding process, a steady rest is generally used. However, considerable skill of the worker is required to use a steady rest. Therefore, we developed a new traverse grinding method without any steady rest. In this method, the elastic deformation of a workpiece was kept constant by controlling the traverse speed of the workpiece. At the middle of the ground workpiece, where the elastic deformation increased easily, the traverse speed was slowed down. However, this method had a longer grinding cycle time because the average traverse speed decreased compared to that of the conventional method. To shorten the cycle time, the peripheral speed of the grinding wheel was increased to decrease the normal grinding force. Basic grinding experiments were carried out under several grinding conditions by changing the peripheral speed of the wheel. From these grinding experiments, it was confirmed that the normal grinding force and the form error of the ground workpiece decreased as the peripheral wheel speed increased. By using results obtained from basic experiments, grinding experiments involving changes in the traverse speed were carried out at two peripheral wheel speeds. The grinding cycle time was reduced successfully by increasing the peripheral wheel speed without an increment in the form error of the ground workpiece. Furthermore, a form error was observed at the end of the workpiece where the grinding wheel traveled away from the workpiece. The form error occurred because the normal grinding force decreased rapidly when the contact length between the workpiece and the wheel was decreased at the end of the workpiece. To prevent rapid changes in the normal grinding force, the traverse speed of the workpiece was increased at the end of the workpiece. By using this method, a ground workpiece with high form accuracy was obtained.

Metal-free magnetic mixed micelles (mean diameter: 16 nm) composed of biocompatible surfactant Tween 80 and hydrophobic pyrrolidine-N-oxyl radical were prepared by mixing them in phosphate-buffered saline. The magnetic mixed micelles were characterized by dynamic light scattering and small angle neutron scattering measurements. The stability of the micelles is found to depend on the length of alkyl side chain in the nitroxide compounds and degree of unsaturation in the hydrophobic chain in the surfactant. The size of the mixed micelle can be tuned by changing the molar ratio of Tween 80 and nitroxyl radical. In view of theranostic application of the micelle, the cytotoxicity and stability in a physiological environment was investigated; the mixed micelle exhibited no cytotoxicity, high colloidal stability and high resistance towards reduction by large excess ascorbic acid. The in vitro and in vivo magnetic resonance imaging (MRI) revealed sufficient contrast enhancement in the proton longitudinal relaxation time (T 1) weighted images. In addition, hydrophobic fluorophores and an anticancer drug are stably encapsulated in the mixed micelles and showed fluorescence (FL) upon reduction by ascorbic acid and cytotoxicity to cancer cells, respectively. For example, the paclitaxel-loaded mixed micelles efficiently suppressed cancer cell growth. Furthermore, they were found to give higher MRI contrast (higher r 1 value) in vitro than the micelles without paclitaxel. The magnetic mixed micelles presented here are promising theranostic agents in nanomedicine due to their high biocompatibility and high resistivity towards reduction as well as functioning as a drug carrier in therapy and MR or FL imaging probe in diagnosis.

The use of sulfur dioxide (SO2) as an antimicrobial in winemaking is a well-established, common practice. Although much is known about the antimicrobial effects of SO2 at single concentrations, little is known about its effects on microbial growth dynamics across a range of concentrations or when used in conjunction with yeast inoculation. Using high-throughput marker-gene sequencing, we investigated the cumulative impacts of yeast inoculation and SO2 treatments across a broad concentration range (0 to 150 mg/L SO2) on the bacterial and fungal communities in wine fermentations. Our results indicated a dose-dependent effect of SO2, with lactic acid bacteria and Gluconobacter proliferating in fermentations with

Background Endotoxin (Et) in the portal vein blood is processed by the hepatic reticuloendothelial system. Thus, it is possible that the Et kinetics of the peripheral venous blood may be useful as a biological index that can be used to evaluate liver function. In this study, we measured Et using the endotoxin activity assay in peripheral venous blood during living donor liver transplantation (LDLT), to study its clinical significance. Methods Subjects were 17 patients who underwent LDLT. In the perioperative peripheral venous blood, was measured Et activity (EA) using the endotoxin activity assay at 1 or 2 d before LT, and then on 1, 5, 7, 14, and 21 postoperative days. Results Patients with infections had significantly higher EA levels compared with those without complications before LDLT and 14 postoperative days (P = 0.038 and 0.027, respectively). The average EA level of patients with infections and without complications before LT was 0.22 and 0.08, respectively (P = 0.038). Patients with an EA level higher than 0.20 before LDLT had a significantly longer period of hospitalization compared with those without complications (P = 0.038). Conclusions A preoperative EA level more than 0.20 is a high risk factor for post-transplant infection and a prolonged period of hospitalization.

Background Acute cellular rejection (ACR) is a common cause of morbidity following liver transplantation. Several reports have evaluated the predictive value of peripheral blood eosinophilia as a simple noninvasive diagnostic marker for ACR. This study examined whether the relative eosinophil counts (REC) predicted ACR in pediatric living donor liver transplantation (LDLT). Methods One hundred three patients underwent LDLT between May 2001 and December 2007. ACR were diagnosed based on the pathological findings. Results The incidence of ACR was 46.6% (48/103); ACR was diagnosed an average of 13.5 days after LDLT. The average REC at 4 and 2 days before the onset ACR (n = 39) within 30 postoperative day (POD) was 4.3% and 7.3%, respectively, and 9.0% at the onset. Patients with ACR showed significantly higher levels of REC compared with those free of ACR ( P = .039). REC thresholds of 10% at POD 7 displayed a sensitivity and specificity of ACR detection of 80% and 75%, respectively. Moreover, the accumulated morbidity ratio of ACR within 30 POD was significantly higher with REC >10% at POD 7 ( P = .007). Conclusion ACR within POD 30 should be considered when REC is >10% at POD 7 after LDLT.

Liver transplantation (LT) has been adopted as a radical treatment for ornithine transcarbamylase deficiency (OTCD), yielding favorable outcomes. Despite the fact that it is an inheritable disease, a blood relative who is heterozygous for the disorder must sometimes be used as a liver donor for living donor LT. There is ongoing discussion regarding the use of heterozygous donors, however, to our knowledge, no cases where donation was determined based on the Ornithine transcarbamylase (OTC) activity before LT have been reported. Between May 2001 and April 2011, 17 patients were indicated for living donor LT because of OTCD at our facility. There were three cases with heterozygous donor candidate (17.6%). All heterozygous candidates underwent a liver biopsy to measure their OTC activity before LT and made efforts to secure the safety of the both donor and recipient. Two of 3 candidates had headaches sometimes, and their activity was less than 40%, and thus they were not employed as the donor. One candidate with 104.4% activity was employed, yielding favorable outcomes. Our current experience supported the effectiveness of our donation criteria, however it is necessary to collect sufficient data on a large number of patients to confirm the safety of the procedure.

Background Excessive portal pressure at an early stage after living-donor liver transplantation (LDLT) can damage sinusoidal endothelial cells and hepatocytes through shear stress leading to graft failure, or hepatic arterial complications due to low hepatic artery flow from a hepatic arterial buffer response. We encountered a case in which excessive portal vein flow was observed from an early stage after pediatric LDLT. The hepatic artery flow decreased due to a hepatic arterial buffer response. Case report A 6-month-old boy with biliary atresia showed excessive portal vein flow early after LDLT with a decreasing hepatic artery flow without anastomotic stenosis from postoperative day 3. The PV flow gradually exhibited a decrease at approximately postoperative day 8 and, similtaneously, hepatic artery flow exhibited improvement. Conclusion Because excessive portal pressure after LDLT is reversible, it has been suggested that it may be possible to prevent the progress of hepatic arterial complications if temporary portal pressure modulation can be performed for cases among the high-risk group for hepatic arterial complications.

BACKGROUND At the present time, indications of liver transplantation (LT) for jaundice-free biliary atresia (BA) patients include intractable cholangitis, portal hypertension and pulmonary vascular disorders. However, the timing of LT remains unclear. In the current study, we describe the therapeutic strategies for jaundice-free BA patients. MATERIAL/METHODS 129 BA patients were undergone LDLT between May, 2001 and April, 2010 in the Department of Transplant Surgery, Jichi Medical University, Japan. RESULTS The indications of LDLT for jaundice-free BA patients was 30 patients (30/129, 23%), and included portal hypertension (16 patients, 53%). Among the 16 patients with portal hypertension, there were 7 patients (7/16, 23%) in which uncontrollable gastrointestinal bleeding was an indication of LDLT. There were 5 patients (5/7; 71%) in which bleeding sites were not identified, and 3 patients (3/7; 43%) in which supportive treatments against collateral vessels were performed as a previous treatment. CONCLUSIONS Even in jaundice-free BA patients, after supportive treatments for portal hypertension are performed, it is necessary to assess the esophageal and gastrointestinal varices regularly and to also prepare for LT simultaneously because there is a probability of the complication of uncontrollable gastrointestinal bleeding.

Redox-controlled backbone dynamics in cytochrome c (Cyt c) were revealed by 2D 15N NMR relaxation experiments. 15N T1 and T2 values and 1H-15N NOEs of uniformly 15N-labeled reduced and oxidized Cyt c were measured, and the generalized order parameters (S2), the effective correlation time for internal motion (taue), the 15N exchange broadening contributions (Rex) for each residue, and the overall correlation time (taum) were estimated by model-free dynamics formalism. These dynamic parameters clearly showed that the backbone dynamics of Cyt c are highly restricted due to the covalently bound heme that functions as the stable hydrophobic core. Upon oxidation of the heme iron in Cyt c, the average S2 value was increased from 0.88+/-0.01 to 0.92+/-0.01, demonstrating that the mobility of the backbone is further restricted in the oxidized form. Such increases in the S2 values were more prominent in the loop regions, including amino acid residues near the thioether bonds to the heme moiety and positively charged region around Lys87. Both of the regions are supposed to form the interaction site for cytochrome c oxidase (CcO) and the electron pathway from Cyt c to CcO. The redox-dependent mobility of the backbone in the interaction site for the electron transfer to CcO suggests an electron transfer mechanism regulated by the backbone dynamics in the Cyt c-CcO system.

The root parasitic plants Orobanche spp. (broomrapes) seriously affect agricultural production. A visualization and quantitative analytical method for the interception of nutrients was established using a positron-emitting tracer imaging system and 13NO− 3. By using this analytical method that involves volume normalization with 18F− images, the nitrogen nutrient interception ratio of the Orobanche spp. was calculated to be 73.6 ± 3.9% in a host–parasite system of red clover (Trifolium pratense L.).

Hypocholinesterasemia is often observed clinically, especially in various liver diseases. Not well known, however, is the fact that some patients have a hereditary BChE variant. Little has been reported on liver transplants associated with this hereditary BChE variant. Furthermore, no cases have been reported of a LDLT involving hereditary BChE variant that had been diagnosed preoperatively. A 23-month-old girl who had had a failed Kasai operation for biliary atresia underwent a liver transplant using as a graft her father's lateral segment. Preoperatively, she had been diagnosed with hypocholinesterasemia. As the donor, her father had undergone a preoperative examination, during which he was found to also have hypocholinesterasemia. DNA sequencing revealed that both had the hereditary BChE variant. The unique mutation caused a frame-shift mutation. Variant K was also detected. The patient was discharged 143 days after the operation and has had no problems with immunosuppression since. In conclusion, we report that the hereditary BChE variant is not a contraindication for either transplantation or living liver donation.

Gas injection is generally practiced in refining processes for metallurgy. The expectances for the gas injection involve the homogenization of bath temperature and compositions, and removal of second phase and dissolved impurities from molten metals.It is widely recognized that inclusions in steel cause defects to the products. Therefore, it is important to remove inclusions from liquid steel. Solid inclusions such as alumina and silica are not wetted by the liquid steel and can be removed by attachment to gas bubbles. The attachment and removal by bubble flotation is expected as one of promising techniques on inclusion removal in liquid steel. The aim of the present work is to investigate the effects of some factors such as particle diameter, agitation speed and particle contact angle to removal rate. In this study, water model experiments have been done under turbulent flow condition using a mechanically agitated vessel. It was confirmed that the removal rate of suspended particles in liquid shows first order kinetics until 4 min in the initial stage. Furthermore, it was found that the collision frequency function has decreased with increasing agitation speed and with decreasing contact angle.

The photoassimilate flow in an intact plant stem was imaged in real-time and its dynamics was quantitatively described using the Positron Emitting Tracer Imaging System (PETIS). Radioactive 11CO2 was fed to a leaf of an intact broad bean (Vicia faba L.) plant, together with air containing an ambient concentration of non-radioactive carrier CO2 gas. Movies of flow of the 11C-labeled photoassimilates in the plant body were captured with PETIS. Here we demonstrate that the average flow speeds and the distribution ratios of photoassimilates in the respective nodes and internodes of the observed stem can be estimated by the transfer function analysis, one of the mathematical modeling methods. We also estimated the changes in the spatial distribution of the average flow speeds in the same stem when the fed leaf was exposed to enriched carrier CO2 gas.

We have investigated the usefulness of hypocotyl (cultured on N1B2 medium) and cotyledon explants (on CR medium: Murashige and Skoog (MS) medium, 3% sucrose, 20 µm benzylaminopurine, pH 5.8) for the regeneration of shoots of the Korean radish ‘Jin Ju Dae Pyong’. The importance of ethylene (indirectly), polyamines and gelling agent were studied in both media. Although the addition of ethylene-inhibitors and silver nitrate to the culture media were beneficial towards shoot regeneration and agar-based treatments (0.8% w/v) being superior in shoot production compared to agarose treatments (0.4% w/v), both explants responded differently in culture. Hypocotyls cultured in the presence of silver nitrate or aminoethoxyvinylglycine (AVG) regenerated significantly (p < 0.05) more shoots compared to N1B2 medium alone; supplementation of 20 µM AVG to N1B2 medium gave optimal shoot production (40% of explants regenerating shoots). The addition of 10 µM AVG to CR medium produced maximum shoot regeneration from cotyledon explants (60% producing shoots). Plants derived from 3-month-old cultures produced greater seed weights, larger leaves and greater genetic variability (50–80% of cells having 20–40 chromosomes) compared to seed-derived (85–90% diploid) and plants from 1-month-old cultures (78–88% diploid). Our results show, that if prolonged culture of explants is avoided, a large number of phenotypically-normal plants can be produced, which in turn, could be utilized in the genetic improvement of radish.

A sensitive electrochemical measurement system for hydroxyl radical ( OH) was developed using enzyme-catalyzed signal amplification. In the presence of 2,6-xylenol as a trapping agent, glucose as a substrate, and pyrroloquinoline quinone-dependent glucose dehydrogenase (PQQ–GDH) as a catalyst, the amperometric signal of the trapping adduct 2,6-dimethylhydroquinone (DMHQ) produced by the hydroxylation of 2,6-xylenol was able to be amplified and detected sensitively. The limit of detection (signal/noise [ S / N ] = 3) for DMHQ was 1 nM. There was no significant interference from urate and other oxidizable compounds in the reaction mixture at the applied potential of 0 V versus Ag/AgCl. This method was employed to observe the OH generation induced by the xanthine–xanthine oxidase (XO) system. The reaction rates of the DMHQ production induced from the xanthine–XO system in the presence and absence of various Fe(III) complexes and proteins were compared. Those with a free coordination site on the Fe atom effectively enhanced the OH generation.

Male-associated DNA sequences were analysed in hemp (Cannabis sativa L.), a dioecious plant with heteromorphic sex chromosomes. A male-associated DNA sequence in C. sativa (MADC1) and its flanking sequence encoded a reverse transcriptase that was strongly homologous to those of LINE-like retrotransposons from various plants and other organisms, as well as another open reading frame (ORF). Fluorescence in situ hybridization (FISH) with MADC1 as probe, which yielded strong signals specific for male genomic DNA in gel blot analysis, generated a clear doublet signal at the end of the long arm of the Y chromosome. FISH using pachytene chromosomes of pollen mother cells at meiotic prophase I revealed that pairing of X and Y chromosomes occurred at the short arm of the Y chromosome where MADC1 was not present. Furthermore, FISH using extended DNA fibers, with MADC1 and its flanking DNA as probes, revealed that 100 to 200 copies of the retrotransposon were located in tandem on the Y chromosome. These results support the hypothesis that accumulation of a specific LINE-like retrotransposon at the terminal region of the long arm of the Y chromosome might be one cause of heteromorphism of sex chromosomes.

The sexual pattern of the nine species of labrid fishes collected at the Kuchinoerabu-jima, Kagoshima Prefecture, Japan is described. Histological evidence shows that all nine species are protogynous hermaphrodites. These are: Anampses geographicus, Thalassoma quinquevittata, T. purpureum, Stethojulis trilineata, Halichoeres scapularis, Hologymnosus annulatus, Xyrichtys dea, X geisha, and Epibulus insidiator. Of these, T. purpureum, H. annulatus are diandric. Diandry can not be confirmed in the remaining species because sufficient sample sizes of each species could not be collected at this site.

A dioecious plant, Cannabis sativa has two sex chromosomes (X and Y). The genome sizes of the diploid female and male plants were determined to be 1636 and 1683 Mbp, respectively, by flow cytometry. By the karyotype analysis, the X and Y chromosomes were found to be submetacentric and subtelocentric, respectively. The Y chromosome had the largest long arm with a satellite in the terminal of its short arm. Conspicuous condensation was specifically observed in the long arm and satellite of the Y chromosome during the prometaphase to metaphase stages. These results indicate that the Y chromosome, especially in its long arm, specifically differentiates in Cannabis sativa and might contribute to the sex determination.

Information about the transfer of antiarrhythmic drugs into milk is available. However, results for the same drug sometimes vary. This variation may be due to factors such as interpatient variation of drug transfer or the small number of patients in the studies. Equations are useful for predicting drug transfer into milk [1]. Pilsicainide is a new class I antiarrhythmic drug used in Japan. We collected serial specimens of plasma and milk after a single dose of three different antiarrhythmic drugs, including pilsicainide, in a healthy mother. We also compared the results with an equation for predicting the milk/plasma ratio. One healthy nursing mother (one of the authors, M.W.; body weight 56 kg) took a single oral dose of either pilsicainide (50 mg), disopyramide (100 mg) or propafenone (150 mg) after a 12-h fast. The wash-out period between each dose was 4 weeks and the order of the section was randomly chosen. She was prohibited from breast feeding on trial days. Blood and milk samples were collected at 1, 2, 3, 4, 6, and 12 h after each dosing. Breast milk samples were collected by manual expression and milk pH was measured anaerobically. For sampling milk, breasts were emptied of milk at each time point and a stirred aliquot was taken for analysis. Protein binding of each drug was measured after microultrafiltration. Concentrations of pilsicainide, propafenone, 5-(OH)propafenone and disopyramide were measured using high-performance liquid chromatography [2, 3] or enzyme immunoassay [4]. The detection limit was 0.06 µg ml−1, 10 ng ml−1, 10 ng ml−1, and 0.07 µg ml−1, for pilsicainide, propafenone, 5-(OH)propafenone and disopyramide, respectively. The milk/plasma ratio of each drug (M/Pobs) was calculated by the ratio of AUC0−12 h for milk and plasma, obtained by the trapezoidal method. Pharmacokinetic parameters of pilsicainide after a single oral administration were estimated using the least square method applied to a one-compartment open model with first-order absorption and elimination. The maximum and minimum serum concentrations of pilsicainide at steady state were calculated by the estimated pharmacokinetic parameters using computer software. The body weight-adjusted relative infant dose of the drug was calculated from the assumption that a baby takes 150 ml kg−1 day−1 of milk. Finally, we estimated the M/P ratio (M/Ppred) according to the previously reported equations of Begg [1], and compared the results with actual M/Pobs. The Ethics Committee of the Medical School approved the protocol. The pilsicainide concentration in milk was higher than that in plasma at each observation point, whereas disopyramide and propafenone concentrations in milk were lower than those in plasma (Table 1). Estimated Cmax/Cmin at steady state, M/Pobs, protein binding, milk pH, octanol/water partition coefficient at pH 7.2 and pKa used for the prediction are also shown. Table 1 Pharmacokinetic profiles of three antiarrhythmic drugs The value of M/P for disopyramide varies widely (from 0.4 to 0.9) among reports [5–7]. Because the protein binding ratio of disopyramide is nonlinear with respect to concentration [8], this difference might be due to the different concentrations of the drugs used in these reports. The M/Pobs of pilsicainide is highest among the three drugs and concentrations were detected at 12 h in both plasma and milk after a single dosing. This is the first report on the M/P ratio of pilsicainide. AUC0−12 h of pilsicainide in milk was 4.7 µg h−1 ml−1. If we assume that a baby takes 150 ml kg−1 day−1 of milk, the estimated amount of ingested drug is 0.05875 mg, which means that the body weight-adjusted relative infant dose is 7% (Table 1). Because 10% is the notionally accepted cut-off for an average drug, this value for pilsicainide is just within the accepted range. The M/Pobs for propafenone was lowest among the three in this study. There is one report for propafenone [9] and the value is comparable to that reported in this study. The equation of Begg gave a close M/Ppred for pilsicainide. However, since there was only one subject in this study, further studies should be conducted with a larger number of subjects to confirm these findings. We did not collect samples more than 12 h after dosing, which is a limitation of this study. AUC0−12 h was between 89 and 100% of AUC0–8 for the three drugs. We did not measure N-monodealkylated disopyramide, which has higher cholinergic activity than the parent compound, in this study. Accumulation of this metabolite into breast milk is reported [5], which might also have occurred in our study. Although the relative infant dose for disopyramide is low, careful monitoring is needed in the clinical setting. In conclusion, we report here for the first time the drug transfer of pilsicainide after a single dosing in a healthy mother. We also evaluated transfer of disopyramide and propafenone in an identical mother. Begg's equation is effective for predicating the M/P-value, especially for pilsicainide. These observations may be useful for the treatment of arrhythmia in patients who are breast feeding.

We discuss the influence of apodized pupils with central obscuration on the incident Gaussian beam. Curves of transverse, as well as of longitudinal irradiance distributions are presented for some apodizers and for several values of the obscuration ratio.

From the viewpoint of energy security and global warming issues, the integrated coal gasification combined cycle (IGCC), a highly efficient coal-based power generation system, is regarded as an effective clean coal technology that is close to commercialization. Combination of the IGCC with CO2 capture and storage technology has been planned; this concept further attracts people looking for ways to reduce CO2 emissions. As one of the leading company in the field of IGCC reducing CO2 emission, Mitsubishi Heavy Industries, Ltd. (MHI) is now in a position of actively undertaking its realization with an integration of two stage entrained bed air-blown coal gasifier, which achieves the highest net plant efficiency by using air as the gasification agent, and high temperature combustion turbines. This paper will discuss the latest activities MHI have been taking at the 250 MW IGCC demonstration plant in Nakoso, Japan and air-blown IGCC commercial plant projects using high temperature combustion turbines. As a future vision to the carbon constrained world, MHI’s perspective on upcoming technologies like higher temperature combustion turbines represented by “J”-type, 1,700°C-class will be introduced.

There are no objective and concrete guidelines for the management of Ornithine transcarbamylase deficiency (OTCD). Based on previous findings, we hypothesized that patients with OTCD have a low Ornithine transcarbamylase (OTC) activity in the liver, and therefore it would be better to determine the appropriate indications and optimal timing for liver transplantation (LT) based on the OTC activity. However, few data have so far been accumulated on the OTC activity in cases that are indicated for LT. The purpose of the present study was to clarify the OTC activity in cases that were indicated for LT. This study involved thirteen children with OTCD (8 males and 5 females) who underwent LT, and two females with OTCD who did not require LT. The OTC activity of the neonatal onset type ranged from 0% to 7.2%, while that of the late onset type who underwent LT ranged from 4.4% to 18.7%. The OTC activity of the late onset type which did not require LT was 33-38% based on a preoperative needle liver biopsy. Some late onset patients that underwent LT, showed an activity that was as low as that observed in the neonatal onset cases. This is the first report to show the results of measuring the OTC activity for serial OTCD cases indicated for LT. OTC activity might be an indicator to determine the indications for and the timing of LT in the late onset type, however, further investigations are necessary.

The final interprotein electron transfer (ET) in the mammalian respiratory chain, from cytochrome c (Cyt c ) to cytochrome c oxidase (C c O) is investigated by 1 H- 15 N heteronuclear single quantum coherence spectral analysis. The chemical shift perturbation in isotope-labeled Cyt c induced by addition of unlabeled C c O indicates that the hydrophobic heme periphery and adjacent hydrophobic amino acid residues of Cyt c dominantly contribute to the complex formation, whereas charged residues near the hydrophobic core refine the orientation of Cyt c to provide well controlled ET. Upon oxidation of Cyt c , the specific line broadening of N-H signals disappeared and high field 1 H chemical shifts of the N-terminal helix were observed, suggesting that the interactions of the N-terminal helix with C c O are reduced by steric constraint in oxidized Cyt c , while the chemical shift perturbations in the C-terminal helix indicate notable interactions of oxidized Cyt c with C c O. These results suggest that the overall affinity of oxidized Cyt c for C c O is significantly, but not very much weaker than that of reduced Cyt c . Thus, electron transfer is gated by dissociation of oxidized Cyt c from C c O, the rate of which is controlled by the affinity of oxidized Cyt c to C c O for providing an appropriate electron transfer rate for the most effective energy coupling. The conformational changes in Lys13 upon C c O binding to oxidized Cyt c , shown by 1 H- and 1 H, 15 N-chemical shifts, are also expected to gate intraprotein ET by a polarity control of heme c environment.

Cytochrome P450s (P450s) contribute to carcinogenesis by activating procarcinogens and also metabolize anti-cancer drugs. The activity and protein levels of P450s are important in cancer risk and in cancer therapy. In this study, we found that overexpression of CYP3A4 induced growth of a human hepatoma cell line, Hep3B. Overexpression of CYP2D6, by comparison, decreased cell growth. An inhibitor of CYP3A4, ketoconazole, significantly suppressed the growth of Hep3B cells overexpressing CYP3A4, but an inhibitor of CYP2D6, quinidine, did not restore Hep3B cell growth to baseline levels. Overexpression of CYP3A4 increased the production of reactive oxygen species, but this was not the cause of the CYP3A4-induced growth. Previously, we showed that CYP3A4 can produce epoxyeicosatrienoic acids (EETs) from arachidonic acid. The CYP3A4-enhanced cell growth was attenuated by a putative EET receptor antagonist, 14,15-EEZE. CYP3A4 promoted progression of the cell cycle from the G1 to the S phase. CYP3A4 also induced a hypoxic response of Hep3B cells, detected as enhanced erythropoietin gene expression (a typical hypoxic response). The cell growth promoted by CYP3A4 was inhibited by PI3K inhibitor LY294002. These results suggest that CYP3A4 plays an important role in tumor progression, independent of the activation of carcinogens and metabolism of anti-cancer drugs.

Endotoxin (Et) in the portal vein blood is processed by the hepatic reticuloendothelial system, and therefore, it is possible that the hepatic clearance of Et may become a biological index for liver function. In this study, Et levels of preoperative peripheral and portal vein blood at the time of liver transplantation (LT) were measured in order to study the meaning.The study population comprised 19 patients in whom pediatric living donor LT was performed. In the preoperative peripheral and the portal vein blood at the time of LT, we measured Et activity (EA) by the Et activity assay (EAA) and the Limulus amebocyte lysate (LAL) method.The preoperative peripheral vein blood showed a low EA in all cases. In the EA of the peripheral and the portal vein blood, the latter showed a significantly high level (p = 0.049). With the LAL method, 5.3% (2/38) of patients were positive for Et.The EAA is considered to be superior to the LAL method for the detection of Et, even in low endotoxinemia, and is also capable of elucidating the Et kinetics by accurately reflecting hepatic clearance.

Stabilization of protein structures and protein–protein interactions are critical in the engineering of industrially useful enzymes and in the design of pharmaceutically valuable ligands. Hydrophobic interactions involving phenylalanine residues play crucial roles in protein stability and protein–protein/peptide interactions. To establish an effective method to explore the hydrophobic environments of phenylalanine residues, we present a strategy that uses pentafluorophenylalanine (F5Phe) and cyclohexylalanine (Cha). In this study, substitution of F5Phe or Cha for three Phe residues at positions 328, 338, and 341 in the tetramerization domain of the tumor suppressor protein p53 was performed. These residues are located at the interfaces of p53–p53 interactions and are important in the stabilization of the tetrameric structure. The stability of the p53 tetrameric structure did not change significantly when F5Phe-containing peptides at positions Phe328 or Phe338 were used. In contrast, the substitution of Cha for Phe341 in the hydrophobic core enhanced the stability of the tetrameric structure with a Tm value of ∼100°C. Phe328 and Phe338 interact with each other through π-interactions, whereas Phe341 is buried in the surrounding alkyl side-chains of the hydrophobic core of the p53 tetramerization domain. Furthermore, high pressure-assisted denaturation analysis indicated improvement in the occupancy of the hydrophobic core. Considerable stabilization of the p53 tetramer was achieved by filling the identified cavity in the hydrophobic core of the p53 tetramer. The results indicate the status of the Phe residues, indicating that the “pair substitution” of Cha and F5Phe is highly suitable for probing the environments of Phe residues. © 2011 Wiley Periodicals, Inc. Biopolymers 95: 410–419, 2011

Flow characteristics of a miscible gravity current on a slope are investigated. A number of important flow properties involved are quantified experimentally as functions of bottom slopes. The study is directed toward the estimation of an entrainment coefficient in the front part of the flow.

Aims To examine whether bile acids such as ursodeoxycholic acid (UDCA) and chenodeoxycholic acid (CDCA) can influence the absorption of nitrendipine, a highly lipophilic calcium channel blocker. Methods Six healthy subjects received nitrendipine (10 mg) with and without UDCA (50 mg) and CDCA (200 and 600 mg) with an interval of 1∼2 weeks between study phases. Results Bile acids decreased the Cmax (ng ml−1) [control 10.9 ± 5.8 (mean± s.d.), UDCA 5.0 ± 4.7 (95% confidence interval for difference; 3.9, 7.8, P = 0.0006), CDCA (600 mg) 5.0 ± 3.9 (2.6, 9.2, P = 0.0059)] and AUC (ng ml−1 h) [(control; 60 ± 36, UDCA 15 ± 13 (20, 73, P = 0.0064), CDCA (600 mg) 19 ± 19 (21, 63, P = 0.0038)] of nitrendipine, while elimination half-life remained unchanged. Conclusions These results suggest that the amount of nitrendipine absorbed was decreased when the drug was administered with UDCA and CDCA.