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Recent reports have described the practicality of laparoscopic intragastric surgery (l-IGS) as an alternative for resecting submucosal tumors (SMTs) near the esophagogastric junction (EGJ), where excision using an exogastric approach would be difficult. However, even using IGS to perform a full-thickness resection of SMTs that are in or extremely close to the EGJ is very difficult to do safely and avoid disrupting or causing stenosis of the EGJ, without advanced experience. This study retrospectively examined the usefulness of l-IGS for gastric SMTs located in or extremely close to the EGJ. Fourteen patients with gastric SMTs < 2 cm of the EGJ and underwent l-IGS were eligible for this study. We examined the tumor location, operative time, intraoperative hemorrhage, degree of deformation, gastroesophageal reflux disease, perioperative complications, and recurrence. Furthermore, we compared patients with tumors in the EGJ with those with tumors near the EGJ and patients in whom three-port l-IGS was performed with those who underwent single-incision laparoscopic surgery. The average tumor size, operative time, intraoperative hemorrhage, and postoperative hospitalization of the 14 patients were 30.9 ± 21.3 mm, 125.2 ± 31.1 min, 30.7 ± 103.3 mL, and 9.2 ± 3.1 d, respectively. No differences in these parameters according to the type of l-IGS or tumor location were observed. All patients underwent l-IGS without complications and were free from EGJ deformation or esophagitis. We believe that l-IGS is useful for gastric SMTs located < 2 cm of the EGJ as it can be safely performed for difficult tumor locations and does not cause deformation of the EGJ.

Olanzapine is an antipsychotic agent with species-dependent pharmacokinetic profiles in both humans and animals. In the present study, the metabolic profiles of olanzapine in vitro and in vivo were compared in non-transplanted immunodeficient NOG-TKm30 mice and chimeric mice with humanized livers (hereafter humanized-liver mice). Hepatic microsomal fractions prepared from humanized-liver mice and humans mediated olanzapine N10-glucuronidation, whereas fractions from cynomolgus monkeys, marmosets, minipigs, dogs, rabbits, guinea pigs, rats, CD1 mice, and NOG-TKm30 mice did not. The olanzapine N10-glucuronidation activity in liver microsomes from humanized-liver mice was inhibited by hecogenin, a human UDP-glucuronosyltransferase (UGT) 1A4 inhibitor. In addition, hepatocytes from humanized-liver mice suggest that olanzapine N10-glucuronidation was a major metabolic pathway in the livers of humanized-liver mice. After a single oral dose of olanzapine (10 mg/kg body weight) to humanized-liver mice and control NOG-TKm30 mice, olanzapine N10-glucuronide isomers and olanzapine N4′-glucuronide were detected only in the plasma of humanized-liver mice. In contrast, the area under the curve for N4′-demethylolanzapine, 2-hydroxymethylolanzapine, and 7-hydroxyolanzapine glucuronide was higher in NOG-TKm30 mice than that in humanized-liver mice. The cumulative excreted amounts of olanzapine N10-glucuronide isomers were high in the urine and feces from humanized-liver mice, whereas the cumulative excreted amounts of 2-hydroxymethylolanzapine were higher in NOG-TKm30 mice than in humanized-liver mice. Thus, production of human-specific olanzapine N10-glucuronide was observed in humanized-liver mice, which was consistent with the in vitro glucuronidation data. These results suggest that humanized-liver mice are useful for studying drug oxidation and conjugation of olanzapine in humans.SIGNIFICANCE STATEMENTHuman-specific olanzapine N10-glucuronide isomers were generated in chimeric NOG-TKm30 mice with humanized livers (humanized-liver mice), and high UGT1A4-dependent N10-glucuronidation was observed in the liver microsomes from humanized-liver mice. Hence, humanized-liver mice may be a suitable model for studying UGT1A4-dependent biotransformation of drugs in humans.

The March 2011 Fukushima Dai-ichi Nuclear Power Plant accident in Japan released 520 PBq of radionuclides compared to a total release of 5300 PBq from the Chornobyl Nuclear Power Plant accident. Both nuclear accidents resulted in deposition of radiocesium throughout the northern hemisphere, and a plethora of studies have been performed regarding radiocesium (137Cs) behavior. However, few studies have assessed the impact of precipitation on 137Cs deposition in forests. Wide-scale environmental measurements from 2011 and 2016 were used to determine the differences in 137Cs deposition because of precipitation following the Fukushima accident. In areas where wet deposition processes were dominant, dense forests generally had lower ambient dose rates and levels of contamination on forest floors than other stands with fewer stems per hectare in 2011. Similar tendencies were not observed in areas that were primarily subject to dry deposition nor were any trends observed in 2016. 137Cs was retained in dense forest canopies for an extended period regardless of the deposition mode. Additionally, it was found that the initial retention of radionuclides by forest canopies is in general higher for areas with predominantly dry deposition. Incorporation of radiocesium into wood tissues was the same for both wet and dry deposition.

The crystal structures and electron density distributions (EDDs) of Ca-deficient Sc-doped CaTiO3fast oxide-ion conductors, Ca0.97(Ti0.97Sc0.03)O3−δ(CTS3) and Ca0.96(Ti0.9Sc0.1)O3−δ(CTS10), were investigated in the temperature range of 298–1173 K in N2to analyze the effect of composition on the perovskite structure and oxide-ion transport mechanism. CTS3 and CTS10 exhibited orthorhombic Pnmasymmetry in temperature ranges of 298–1173 K and 973–1173 K, respectively, with CTS10 exhibiting lower symmetry and reduction in oxide-ion conductivity below 973 K. The EDDs of CTS3 and CTS10 at 1173 K indicated unique chemical bonds and conduction paths. CTS3 and CTS10 showed covalent bonds between (Ti,Sc) and O1 (or O2) sites. CTS3, with a lower oxide-ion conductivity than that of CTS10, exhibited pseudo-one-dimensional (1D) zig-zag curved conduction paths for oxide-ions along the a-axis, unlike previously reported curved migration paths along the b-axis in CaTiO3, and chemical bonds between Ca and O1 sites, indicating oxide-ion conduction suppression. In CTS10, additional conduction paths were observed along the a-axis, forming three-dimensional (3D) zig-zag curved conduction paths in the ac-plane and along the b-axis, with the weakening of the chemical bonds between the Ca and O1 sites. The oxide-ion conductivity and mobile oxide-ion concentration of CTS10 were 3.6 and 2.0 times those of CTS3, respectively, at 1173 K; the higher oxide-ion conductivity of CTS10 could be attributed to an increase in the mobile oxide-ion concentration and mobility with a 1D to 3D change in the conduction paths and a weakening of chemical bonds between the Ca and O1 sites.

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This paper describes the features and functions of the newly developed Eureka-147 DAB (Digital Audio Broadcasting) receiver. In this paper, we also describe an AFC (Automatic Frequency Control) and a synchronization technique using a DSP (Digital Signal Processor).

Background: Feeding jejunostomy (FJ) is a common treatment to support patients with esophageal cancer after esophagectomy. However, severe FJ-related complications, such as bowel obstruction, occasionally occur. We investigated the ability of our simple, novel FJ technique, the “curtain method,” to prevent bowel obstruction. Methods: In laparoscopic surgery, the main mechanism of bowel obstruction involves torsion of the mesentery accompanied by migration of the intestine across the fixed FJ through the space surrounded by a triangle comprising the ligament of Treitz, fixed FJ, and spleen rather than adhesion. Our “curtain method” involves closure of this triangle zone with omentum, and the appearance of the lifted omentum resembles a curtain. Sixty patients treated with this modified FJ were retrospectively compared with 13 patients treated with conventional FJ in terms of the incidence of bowel obstruction, peritonitis, stoma site infection, and catheter obstruction. Results: From 2013 to 2017, 60 patients underwent esophagectomy and gastric conduit reconstruction accompanied by modified laparoscopic FJ. The median observation period, including the period after tube removal, was 644 days. No FJ-associated bowel obstruction, the prevention of which was the primary aim, occurred in any patient. Likewise, no peritonitis or dislodgement occurred. Eight patients (13%) developed a stoma site infection with granulation. The feeding tube became occluded in 11 patients (18%); however, a new feeding tube was reinserted under fluoroscopy for all of these patients. From 2003 to 2012, 13 patients underwent conventional FJ. The median observation period was 387 days. Three patients (23%) developed bowel obstruction by torsion 71 to 134 days after the first surgery, and all were treated by emergency operations. Other FJ-related complications were not different from those in the modified FJ group. Conclusion: Our simple, novel technique, the “curtain method,” for prevention of laparoscopic FJ-associated bowel obstruction after esophagectomy is a safe additional surgery.

The catalytic enantioselective synthesis of tetrahydrofurans, which are found in the structures of many biologically active natural products, via a transition-metal-catalyzed hydrogen atom transfer (TM-HAT) and radical-polar crossover (RPC) mechanism is described herein. Hydroalkoxylation of nonconjugated alkenes proceeded efficiently with excellent enantioselectivity (up to 94% ee) using a suitable chiral cobalt catalyst, N-fluoro-2,4,6-collidinium tetrafluoroborate, and diethylsilane. Surprisingly, the absolute configuration of the product was highly dependent on the steric hindrance of the silane. Slow addition of the silane, the dioxygen effect on the solvent, thermal dependence, and DFT calculation results supported the unprecedented scenario of two competing selective mechanisms. For the less-hindered diethylsilane, a high concentration of diffused carbon-centered radicals invoked diastereoenrichment of an alkylcobalt(III) intermediate by a radical chain reaction, which eventually determined the absolute configuration of the product. On the other hand, a more hindered silane resulted in less opportunity for a radical chain reaction, instead facilitating enantioselective kinetic resolution during the late-stage nucleophilic displacement of the alkylcobalt(IV) intermediate.

ABSTRACTThis study determined the initial distribution of Fukushima reactor-derived radiocesium fallout in forest areas of Fukushima Prefecture based on analysis of airborne monitoring surveys by the Japanese Ministry of Education, Culture, Sports, Science and Technology (MEXT). The results of the fifth airborne monitoring surveys were compared with the third airborne survey data in order to correct for variation of deposition density due to multiple factors such as weathering processes and measurement uncertainties between two surveys. Finally, the results of the third and the corrected fifth airborne surveys were combined to reconstruct the initial radiocesium fallout map over the whole Fukushima Prefecture area following the Fukushima reactor accident. Our calculation results showed that forest areas accumulated 72% of the total atmospheric input of 137Cs to the land of Fukushima Prefecture. The deposition density in forest areas showed significant variability among municipalities. Nevertheless, forest areas accumulated a large percentage of atmospherically deposited radiocesium in many municipalities. Statistical analysis of the variability in 137Cs deposition density indicated that deposition density varied significantly, even within a municipality. Furthermore, the inconsistency in the probability distributions of measured deposition density among municipalities makes it difficult to obtain a representative initial radiocesium fallout input following the accident. To reduce uncertainty in environmental parameters related to radiocesium transfer in forest environments, we strongly recommend that the deposition density of initial 137Cs fallout at each location is determined by in-situ sampling survey and using the reconstructed 137Cs fallout map presented in this study.

AbstractOwing to their photosynthetic capabilities, there is increasing interest in utilizing cyanobacteria to convert solar energy into biomass. 2-Deoxy-scyllo-inosose (DOI) is a valuable starting material for the benzene-free synthesis of catechol and other benzenoids. DOI synthase (DOIS) is responsible for the formation of DOI from d-glucose-6-phosphate (G6P) in the biosynthesis of 2-deoxystreptamine-containing aminoglycoside antibiotics such as neomycin and butirosin. DOI fermentation using a recombinant Escherichia colistrain has been reported, although a carbon source is necessary for high-yield DOI production. We constructed DOI-producing cyanobacteria toward carbon-free and sustainable DOI production. A DOIS gene derived from the butirosin producer strain Bacillus circulans(btrC) was introduced and expressed in the cyanobacterium Synechococcus elongatusPCC 7942. We ultimately succeeded in producing 400 mg/L of DOI in S. elongatuswithout using a carbon source. DOI production by cyanobacteria represents a novel and efficient approach for producing benzenoids from G6P synthesized by photosynthesis.

A patient with an aortobronchial fistula secondary to a thoracic endovascular aortic repair was successfully treated with a re-thoracic endovascular aortic repair with debranching technique. Five months postoperatively, the aneurysm had shrunk; however, computed tomography revealed air in the thrombo-excluded aortic aneurysmal sac without signs of infection. Because of worsening air finding at the eighth month, we performed a resection of the residual fistula and wrapped the stent graft in a pedicled muscle flap.

Esophageal squamous cell carcinoma (ESCC) is one of the most aggressive cancers and is primarily treated with platinum-based neoadjuvant chemotherapy (NAC). Some ESCCs respond well to NAC. However, biomarkers to predict NAC sensitivity and their response mechanism in ESCC remain unclear. We perform whole-genome sequencing and RNA sequencing analysis of 141 ESCC biopsy specimens before NAC treatment to generate a machine-learning-based diagnostic model to predict NAC reactivity in ESCC and analyzed the association between immunogenomic features and NAC response. Neutrophil infiltration may play an important role in ESCC response to NAC. We also demonstrate that specific copy-number alterations and copy-number signatures in the ESCC genome are significantly associated with NAC response. The interactions between the tumor genome and immune features of ESCC are likely to be a good indicator of therapeutic capability and a therapeutic target for ESCC, and machine learning prediction for NAC response is useful.

Japanese cedar (Cryptomeria japonica) and Japanese cypress (Chamaecyparis obtusa) plantations account for approximately 30% of the total forested area in Japan. Both are arbuscular mycorrhizal trees that leach more NO3−in response to nitrogen (N) deposition than do forests of ectomycorrhizal trees. However, little information is available about the size of N exports from these plantations. The aim of this study was to evaluate nonpoint source N exports from a N‐saturated plantation. We collected stream water samples in base‐flow (25 samples) and storm‐flow conditions (20 events) in a watershed (2.98 ha) where Japanese cypress and Japanese cedar were planted in 1969 (41 yr old). The annual NO3−export was calculated from load–discharge relationships. Atmospheric N deposition was also determined. The stream water contained high NO3−concentrations (160 and 165 μmol L−1during base flow and storm flow, respectively), indicating N saturation in the watershed. High bulk atmospheric N deposition (16.5 kg N ha−1yr−1) could explain the N saturation. There were only small variations in NO3−concentrations in stream water in response to discharge volume, because of the N saturation of the forest ecosystem. Consequently, there were only small errors in estimating annual NO3−exports from the studied watershed. The annual NO3−export was high (36.1 kg N ha−1yr−1), comparable to values reported for agricultural and urbanized areas. These results suggest that N‐saturated forest plantations can become important nonpoint N sources. Our results also suggest that N exports from forest plantations across Japan should be quantified to evaluate nonpoint source N accurately.

Screening of dry eye disease in visual display terminal workers during occupational health examinations: The Moriguchi study: Motoko Kawashima, et al. Department of Ophthalmology, Keio University School of Medicine The aim of this study was to estimate the prevalence of dry eye disease (DED) in Japanese visual display terminal (VDT) workers and identify simple methods for screening DED during occupational health examinations. This study involved 369 (331 men and 38 women; mean age, 44.4 [8.8] years) workers engaged in PC development with approximately 6 hours of VDT use daily. The participants completed questionnaires on dry eye and physical symptoms affecting QoL, and their ability to keep their eyes open for 10 seconds was tested for DED screening. They also underwent assessment of conjunctival and corneal vital staining with lissamine green and fluorescein, measurement of tear film breakup time, and Schirmer's test for DED diagnosis. Sensitivity, specificity, and positive predictive value of the screening methods were assessed. The DED group included 218 (definite, 14; probable, 204) participants. They had markedly high frequencies of 11 dry eye symptoms, especially ocular fatigue (OR, 17.1; 95% CI, 9.9 to 29.7) and dry sensation (OR, 8.2; 95% CI, 3.6 to 18.4), and were more often unable to keep their eyes open for 10 s. With either ocular fatigue or dry sensation as the cutoff criterion, the sensitivity, specificity, and positive predictive value were 89.9, 66.9, and 79.7%, respectively. Approximately 60% of the VDT workers were diagnosed with DED on the basis of diverse eye symptoms affecting their ability to work. The dry eye symptom questionnaire and 10‐s eye‐opening test are simple, noninvasive, and inexpensive methods for screening DED during occupational health examinations.

IMPORTANCE: There are limited reports on the relationship between mucin 5AC (MUC5AC) concentrations in tears, working hours, and the frequency of ocular symptoms in visual display terminal (VDT) users. This investigation evaluated these relationships among patients with dry eye disease (DED) and individuals serving as controls. OBJECTIVE: To determine the relationship between MUC5AC concentration in the tears of VDT users based on the diagnosis of DED and frequency of ocular symptoms. DESIGN, SETTING, AND PARTICIPANTS: An institutional, cross-sectional study was conducted. Participants included 96 young and middle-aged Japanese office workers. Both eyes of 96 volunteers (60 men and 36 women) were studied. Participants working in a company that used VDTs completed questionnaires about their working hours and the frequency of ocular symptoms. Dry eye disease was diagnosed as definite or probable, or it was not present. Tear fluid was collected from the inferior fornix after instillation of 50 μL of sterilized saline. The MUC5AC concentration was normalized to tear protein content and expressed as MUC5AC (nanograms) per tear protein (milligrams). The differences in MUC5AC concentration between DED groups, between VDT working hours (short, intermediate, and long), and between symptomatic and asymptomatic groups were evaluated with 95% CIs based on nonparametric Hodges-Lehmann determination. MAIN OUTCOMES AND MEASURES: Ocular surface evaluation, prevalence of DED, and MUC5AC concentration. RESULTS: The prevalence of definite and probable DED was 9% (n = 9) and 57% (n = 55), respectively. The mean MUC5AC concentration was lower in the tears of VDT users with definite DED than in those with no DED (P = .02; Hodges-Lehmann estimator, −2.17; 95% CI, −4.67 to −0.30). The mean MUC5AC concentration in tears was lower in the group that worked longer hours than in the group that worked shorter hours (P = .049; estimated difference, −1.65; 95% CI, −3.12 to 0.00). Furthermore, MUC5AC concentration was lower in participants with symptomatic eye strain than in asymptomatic individuals (P = .001; estimated difference, −1.71; 95% CI, −2.86 to −0.63). CONCLUSIONS AND RELEVANCE: The data obtained in the present study suggest that office workers with prolonged VDT use, as well as those with an increased frequency of eye strain, have a low MUC5AC concentration in their tears. Furthermore, MUC5AC concentration in the tears of patients with DED may be lower than that in individuals without DED.

AbstractInterception loss (Ei) in forests has been studied widely. However, Eiparameters and modeling as well as spatial patterns of throughfall (TF) in abandoned Japanese cypress (Chamaecyparisobtusa) plantations remain poorly documented. In this study, gross precipitation (PG), stemflow (SF), and TFwere monitored in an unmanaged 32-year-old Japanese cypress stand throughout the 2011 rainy season. Results indicate that PGpartitioning into TF, SF, and Eiwere, respectively, 64.2 ± 3.6, 10.6 ± 0.6, and 25.2 ± 1.1 % of the 880.8 mm cumulative PGfrom 29 rainfall events. Direct throughfall proportion (p) and drainage from the canopy contributed about 14 ± 7 and 50 ± 21 % of the total TFfor the events, respectively. The mean canopy storage capacity (S) was 2.4 ± 0.7 mm. The coefficient of variability (CV) of TFrate decreased asymptotically with increasing PGamount, ranging from 16 to 56 % with median 26 %. The CV of TFrate was not significantly correlated with canopy cover (r= 0.152, P= 0.521, n= 20) and distance from the nearest trunk (r= 0.196, P= 0.408, n= 20). Based on the revised Gash analytical model, the total simulated Eiwas close to the observed, with a general underestimation magnitude of 5.7 %. The Eicomponents were quantified, and most of the interception loss (62.9 %) evaporated during rainfall, while 26.8 % evaporated after rainfall ceased. Climatic and forest structural parameters required by the model were identified and analyzed by sensitivity analysis, implying that the revised Gash analytical model is robust and reliable enough for abandoned Japanese cypress plantations in a maritime climate.

As posttranslational modifications of histones H3 and H4 determine the state of chromatin in cis, these histones should remain attached to template DNA during transcription in order to maintain the state of chromatin. RNA polymerase II itself can transcribe the nucleosome template without changing nucleosome positioning. However, it was uncertain whether Spt6, a highly conserved polymerase-associated histone chaperone, prevents “preexisting” histone molecules from being dissociated from template DNA during transcription. We recently showed that Spt6 prevents transcription-coupled loss of posttranslationally modified histone H3. Taking previous studies into account, we would like to propose here that Spt6 has two fundamentally distinct functions in the regulation of histone modification: one is to act as a platform for histone modifiers and the other is to act as a molecular liaison between histone molecules and template DNA to prevent cotranscriptional dissociation of preexisting histones in order to maintain locus-specific modifications.

Purpose: This study retrospectively evaluated the 12-month outcomes of the Zenith Alpha Abdominal Endovascular graft (Zenith Alpha AAA, Cook, Inc, Bloomington, Indiana), which was launched partially in Japan in March 2019, starting with 9 selected sites.Methods: A retrospective analysis was performed of all endovascular aneurysm repairs (EVAR) for abdominal aortic aneurysms using the Zenith Alpha AAA. Late complications were defined as any aneurysm-related events occurring >30 days after EVAR, including an aneurysm sac enlargement of >5 mm and any reinterventions performed. Endoleaks without sac enlargement or reinterventions were excluded from late complications.Results: During the study period, 79 EVARs using the Zenith Alpha AAA were performed. The mean age was 76.6±6.9 years old, 91.1% of patients were male, and the mean aneurysm diameter was 51.1±7.5 mm. Instructions for use violation was observed in 27 patients (34.2%), most frequently being a severely angulated proximal neck (>60°). There were 4 intraoperative complications, including 2 unintentional partial renal artery coverages caused by the premature anchoring of the exposed suprarenal stent barb to the aortic wall, which was a result of the design change of the top cap deletion. Since it was a partial coverage without flow impairment and since renal stenting was unsuccessful, it was left untreated and had no subsequent renal function impairment. During the mean follow-up of 444±123 days, 74 patients completed 12 months of follow-up. Freedom from late complications at 12 months was 90.8%, which included 2 limb occlusions (2.5%). Of 71 patients with a 12-month computed tomography scan, there was only 1 type 1a endoleak (1.3%), 1 sac enlargement of >5 mm (1.3%), and an aneurysm sac shrinkage of >5 mm was observed in 42.2% of patients. There was no type 3 endoleak during the follow-up.Conclusions: This study demonstrated that the new generation of low-profile Zenith Alpha AAA has satisfactory early clinical outcomes, comparable to those obtained with the conventional Zenith endovascular graft. Long-term follow-up is needed to determine whether these favorable outcomes persist.

We fabricated α-Fe(100)/Nd2Fe14B(001) bilayer films with different thicknesses values of the α-Fe layer on MgO(100) substrates by using an ultra-high-vacuum (UHV) magnetron sputtering system. The exchange coupling constant at the heterointerfaces of α-Fe(100)/Nd2Fe14B(001) was evaluated by analyzing the magnetization curves and the ferromagnetic resonance (FMR) spectra. We observed a significant lower shift of the α-Fe resonance field compared to that of the α-Fe single layer irrespective of the Fe-layer thickness, which suggests the existence of an internal field owing to a positive exchange coupling between the α-Fe(100) and the Nd2Fe14B(001) layers.

We fabricated a model interface system, which consised of a Nd2Fe14B layer with Dy overlayers, in order to study the relationship between the microstructure near the interface and the coercivity. The coercivity Hcof the 1-µm-thick Nd-Fe-B films with a Dy overlayer was enhanced by thermal diffusion of Dy atoms from the film’s surface into the Nd-Fe-B layer. The Nd-Fe-B/Dy films annealed at 700 °C showed a significant increase in the Hcof up to about 17 kOe. The maximum increase in the value of the coercivity ΔHcof the annealed films with a Dy overlayer was approximately 13 kOe. The XRD measurement suggested the existence of rare-earth oxides (NdO and Dy2O3) in the Dy-coated films with high value of the ΔHc.

Tophaceous pseudogout in the knee joint is rare. We report an 82-year-old man who presented with a one-year history of pain and swelling of the right knee joint. Treatment with non-steroidal anti-inflammatory drugs and aspiration of the joint effusion had not been effective. The mass continued to enlarge, and the patient had difficulty walking. Radiographs and computed tomography showed meniscal calcification with an abnormal soft-tissue mass surrounded by calcification. After excision, massive calcified deposits were seen both inside and on the surface of the tophaceous pseudogout. The deposits showed birefringence under polarised light, suggestive of calcium pyrophosphate dihydrate crystals. At the 2-year follow-up, the patient could walk independently without knee pain or swelling, although his range of knee motion was slightly limited due to joint contracture that developed before surgery.

A response regulator, NblR, of the cyanobacterium Synechococcus elongatusPCC 7942 is known to induce expression of the nblAgene, a key factor in phycobilisome degradation (bleaching) under nutrient-deprivation conditions. In this study, we observed phosphorylation-independent regulation of NblR activity. We constructed a mutant strain expressing NblR (D57A), in which a putative phospho-accepting Asp-57 was replaced with Ala residue. Under nitrogen deprivation, this strain exhibited the typical bleaching phenotype observed in wild-type cells. Moreover, in the mutant, the nblAtranscript accumulated at a level similar to that of the wild type. Our results indicate that activation of NblR is independent of phosphorylation, if any, by a cognate histidine kinase. Screening of proteins interacting with NblR by yeast two-hybrid analysis revealed two candidates, MreC and NarB, suggesting a novel mechanism that activates NblR, or other functions of the response regulator.

The six-subunit origin recognition complex (ORC) acts as a landing pad for factors that initiate DNA replication by binding to replication origins. In addition, ORC is involved in other processes such as transcriptional gene silencing and sister chromatid cohesion in Saccharomyces cerevisiae. However, whether these functions of ORC are specific to Saccharomyces cerevisiaeor are shared by the ORC of other organisms is currently unclear. Analysis of two temperature-sensitive alleles of the fifth ORC subunit of Saccharomyces pombe, orc5-H19 and orc5-H37, indicates that Orc5 of Saccharomyces pombehas similar multiple functions to those of Orc5 of Saccharomyces cerevisiae. The orc5-H19 cells were defective in DNA replication initiation, and execution point analysis of this mutant revealed that ORC functions before metaphase to prepare for the initiation of replication in the next cell cycle. The orc5-H37 cells seemed to complete DNA synthesis but were arrested before entering M phase. In both mutants, the rads-chk1 checkpoint was activated to prevent mitosis, suggesting that this checkpoint pathway monitors the functional integrity of ORC. In addition, orc5-H37 cells showed premature separation of sister chromatids, which resulted in cell growth being dependent on the mad2-dependent spindle checkpoint. Consistently, this mutant showed a defect in the loading of Rad21, a cohesin component. Based on these observations, we propose that Orc5 has at least two distinct functions that can be separated genetically. Taken together, our results provide further support for the idea that ORC plays multiple functions during the cell cycle.

We report on the broadening of the operating frequency of a RF splitter widely used in CATV systems. In order to realize this, the relationship between the splitter’s transmission characteristics and design parameters has been investigated using computer-aided engineering. Based on the calculations, a sample device has been fabricated to confirm the theoretical results. It has been found that by selecting the parameters appropriately, excessive loss can be suppressed to less than 1.6dB and 3dB in a frequency range of 20MHz to 2,600MHz in the cases of 2-way and 4-way splitters, respectively.

Abstract PsbP is a membrane extrinsic subunit of Photosystem II (PS II), which is involved in retaining Ca2+ and Cl−, two inorganic cofactors for the water-splitting reaction. In this study, we re-investigated the role of N-terminal region of PsbP on the basis of its three-dimensional structure. In previous paper [Ifuku and Sato (2002) Plant Cell Physiol 43: 1244–1249], a truncated PsbP lacking 19 N-terminal residues (Δ19) was found to bind to NaCl-washed PS II lacking PsbP and PsbQ without activation of oxygen evolution at all. Three-dimensional (3D) structure of PsbP suggests that deletion of 19 N-terminal residues would destabilize its protein structure, as indicated by the high sensitivity of Δ19 to trypsin digestion. Thus, a truncated PsbP lacking 15 N-terminal residues (Δ15), which retained core PsbP structure, was produced. Whereas Δ15 was resistant to trypsin digestion and bound to NaCl-washed PS II membranes, it did not show the activation of oxygen evolution. This result indicated that the interaction of 15-residue N-terminal flexible region of PsbP with PS II was important for Ca2+ and Cl− retention in PS II, although the 15 N-terminal residues were not essential for the binding of PsbP to PS II. The possible N-terminal residues of PsbP that would be involved in this interaction are discussed.

ABSTRACTLoop-mediated isothermal amplification (LAMP), a novel nucleic acid amplification method, was developed for the rapid detection of the major periodontal pathogens Porphyromonas gingivalis, Tannerella forsythia, and Treponema denticola. The LAMP method amplifies DNA with high specificity, efficiency, and rapidity under isothermal conditions using a set of four specially designed primers and a DNA polymerase with strand displacement activity. In this study, we initially designed the primers for LAMP assays to detect these bacteria and evaluated the specificity and sensitivity of these assays. The specificities of the primers for these bacteria were examined using various oral bacteria and various reaction times. The lower detection limits of the 60-min LAMP reaction without loop primers were 1 µg/tube for P. gingivalis, 10 fg/tube for T. forsythia, and 1 ng/tube for T. denticola. Addition of the loop primers for each bacterium improved the detection specificities and sensitivities by several magnitudes. Furthermore, LAMP assays were applied to the rapid detection of these periodontal pathogens in clinical specimens, and the results were compared with those of conventional PCR detection. The results of the LAMP assays corresponded to those of conventional PCR assays. These results indicate that the LAMP assay is an extremely rapid, highly sensitive, specific method. This method is very useful for the rapid detection of periodontopathic bacteria and the diagnosis of periodontal disease.

The calmodulin-binding domain of myristoylated alanine-rich C kinase substrate (MARCKS), which interacts with various targets including calmodulin, actin and membrane lipids, has been suggested to function as a crosstalk point among several signal transduction pathways. We present here the crystal structure at 2 Å resolution of a peptide consisting of the MARCKS calmodulin (CaM)-binding domain in complex with Ca2+-CaM. The domain assumes a flexible conformation, and the hydrophobic pocket of the calmodulin N-lobe, which is a common CaM-binding site observed in previously resolved Ca2+-CaM–target peptide complexes, is not involved in the interaction. The present structure presents a novel target-recognition mode of calmodulin and provides insight into the structural basis of the flexible interaction module of MARCKS.

X-ray and NMR analyses on ribosome recycling factors (RRFs) from thermophilic bacteria showed that they display a tRNA-like L-shaped conformation consisting of two domains. Since then, it has been accepted that domain I, consisting of a three-helix bundle, corresponds to the anticodon arm of tRNA and domain II and a beta/alpha/beta sandwich structure, corresponds to the acceptor arm. In this study, we obtained a RRF from a mesophilic bacterium, Vibrio parahaemolyticus, by gene cloning and carried out an x-ray analysis on it at 2.2 A resolution. This RRF was shown to be active in an in vitro assay system using Escherichia coli polysomes and elongation factor G (EF-G). In contrast, the above-mentioned RRFs from thermophilic bacteria were inactive in such a system. Analysis of the relative orientations between the two domains in the structures of various RRFs, including this RRF from mesophilic bacterium, revealed that domain II rotates about the long axis of the helix bundle of domain I. To elucidate the ribosome binding site of RRF, the peptide fragment (RRF-DI) corresponding to domain I of RRF was expressed and characterized. RRF-DI is bound to 70 S ribosome and the 50 S subunit with an affinity similar to that of wild-type RRF. But it does not bind to the 30 S subunit. These findings caused us to reinvestigate the concept of the mimicry of RRF to tRNA and to propose a new model where domain I corresponds to the acceptor arm of tRNA and domain II corresponds to the anticodon arm. This is just the reverse of a model that is now widely accepted. However, the new model is in better agreement with published biological findings.

Structure–activity relationships for aquatic toxicity were studied using neural networks and linear regression analysis. The structural features contributing to toxicity were identified in molecules exhibiting a level of toxicity greater than that of non-reactive organic molecules. A neural network was trained for the toxicity of non-polar narcotics, polar narcotics, or reactive toxicants. Quantitative structure–activity relationships (QSARs) were developed, relating a molecular aquatic toxicity to its log Pand to a set of 16 structural descriptors based upon the presence of selected structural features. The inclusion of these structural descriptors into a QSAR was found to enhance the correlation of the equation, and thus to improve its ability for predicting aquatic toxicity.

Background:Plasma concentrations of atrial natriuretic peptide (ANP), brain natriuretic peptide (BNP) and cyclic guanosine monophosphate (cGMP) are suitable markers of ’dry body weight’ (DW) in hemodialysis (HD) patients. However, it is still unknown whether these markers can be applied to patients with renal failure and coronary artery disease (CAD). We examined the reliability of these peptides as volume markers in HD patients with CAD. We also assessed the relationship between natriuretic peptides and indices of left ventricular (LV) function. Methods:Plasma concentrations of ANP, BNP and cGMP were determined before and after HD in patients with CAD (group 1, n = 19, mean age 63 ± 12 years) and were compared with those of patients without cardiac disease (group 2, n = 20, age 61 ± 15 years). Using data obtained by cardiac catheterization, we examined the relationship between natriuretic peptides and indices of LV function in HD patients with CAD. Results:Baseline ANP (244 ± 205 pg/ml), BNP (713 ± 928 pg/ml) and cGMP (29.6 ± 21.6 pmol/ml) were significantly higher in group 1 than in 11 healthy volunteers (18.6 ± 9.9 pg/ml, 7.7 ± 7.6 pg/ml, cGMP 8.9 ± 4.9 pmol/ml, respectively). HD significantly reduced plasma ANP (87 ± 75 pg/ml) and BNP (477 ± 702 pg/ml) although they were still above normal control. HD reduced plasma cGMP (7.2 ± 4.5 pmol/ml) to normal values, suggesting the elimination of cGMP across the dialyzers. Baseline levels of ANP, BNP and cGMP in group 2 were less than those of group 1 but higher than the control. HD reduced natriuretic peptides in group 2 to levels lower than those in post-HD group 1. After HD, there was no significant correlation between reductions in body weight and changes in ANP or BNP. Baseline ANP and BNP levels closely correlated with pulmonary artery pressure, pulmonary artery wedge pressure, left ventricular end-diastolic pressure and left ventricular ejection fraction. A significant correlation was observed between BNP levels and the severity of CAD. Conclusion:ANP, BNP and cGMP seem to be a useful markers for fluid overload but not for DW in HD patients with CAD. Plasma ANP and BNP might be useful markers for left ventricular function.

In an effort to explore properties important in hematogenous metastasis of pancreatic adenocarcinoma, we previously demonstrated that tumor‐derived interleukin (IL)‐6 is a crucial factor that conveys resistance to liver metastasis. Here we extend the study to examine a possible vaccination effect of tumor‐derived IL‐6 in T‐cell‐deficient nude mice, as a model for predicting the effect in immune‐compromised patients. We used a pair of IL‐6‐nonproducing and highly producing pancreatic adenocarcinoma cell lines, PCI‐43 and PCI‐43h, respectively. The reaction intensity of anti‐PCI IgG antibodies in host nude mice was maximal 28 days after inoculation of PCI‐43h cells, and remained high thereafter. A fraction of the pancreatic carcinoma cell lines, namely, PCI‐6, ‐10, and ‐43, expressed surface antigenic determinant(s) reactive with the IgG; but the others, PCI‐19, ‐24, ‐55, ‐64, ‐66, ‐68, ‐72, and ‐79, did not. Inoculation of PCI‐43h but not PCI‐43 suppressed growth of simultaneously inoculated PCI‐43, but not PCI‐24 xenografts. In addition, administration of PCI‐43h, but not PCI‐43 suppressed the growth of PCI‐43 that was xenografted 4 weeks later, thus revealing a vaccination effect of IL‐6‐producing PCI‐43h, but not IL‐6‐nonproducing PCI‐43. These data, obtained from T‐cell‐deficient nude mice, suggest an in vivorole for IL‐6 in inducing IgG‐mediated, pancreatic carcinoma‐specific vaccination against a thymus‐independent antigen.

We established peritoneal dissemination‐prone subcultures (PCI‐43p3) using nude mice by a repetitive in vivoselection of intraperitoneally inoculated PCI‐43, a pancreas adenocarcinoma cell line. The subcultures showed upregulated expression of matrix metalloproteinase (MMP)‐9, but not MMP‐2 in culture supernatants. They also produced increased amounts of vascular endothelial growth factor (VEGF), which was not associated with alterations in isoforms of VEGF mRNA. PCI‐43p3 cells attached to cultured mesothelial cell monolayers more readily than did the parent PCI‐43 cells. The angiogenesis inhibiting agent, TNP‐470, at 30 mg/kg was administered to the model mice, resulting in a prominent suppression of the establishment of peritoneal nodules. The suppression was dependent on the duration of TNP‐470 treatment. TNP‐470 treatment significantly suppressed proliferation of tumor cells in disseminated nodules, assessed in terms of immunostaining for proliferating cell nuclear antigen (PCNA). TNP‐470 did not affect the in vitroattachment between PCI‐43p3 and mesothelial cells. The combined data show that anti‐angiogenic treatment profoundly suppresses the in vivoprocess of peritoneal dissemination.

In this paper, system identification and physical parameter estimation of semiconductor exposure apparatus for active vibration control are discussed. Since the semiconductor exposure apparatus has multi-degrees-of-freedom (multi-DOF) motional mechanism, the apparatus must be treated as a Multi-Input-Multi-Output (MIMO) system. In this paper, subspace based state-space system identification (4SID) method is applied, because it can be applied to MIMO systems easily. Effectiveness of the 4SID method to an MIMO system identification problem is examined through experimental data. An estimation method of physical parameters of the semiconductor exposure apparatus based on the identified models is also presented. Furthermore, a system identification device is described which is developed for modeling of the semiconductor exposure apparatus. It measures input and output data for identification and executes system identification algorithms based on MATLAB commands.

In glomerular hypertension, mesangial cells (MC) are subjected to at least two physical forces: mechanical stretch and high transmural pressure. Increased transmural pressure, as well as mechanical stretch, promotes MC proliferation, which may enhance glomerulosclerosis. The exact mechanism of this effect is not fully understood. We examined the effects of transmural pressure alone on cell proliferation and DNA synthesis and investigated the role of platelet-derived growth factor (PDGF) and basic fibroblast growth factor (bFGF), candidates for mediation of glomerular diseases, in the pressure-induced events. Pressure was applied to cultured MC placed in a sealed chamber using compressed helium gas. Application of pressure resulted in a time-dependent (∼2 h) and pressure level-dependent (∼80 mmHg) increase in cell number (1.4-fold) and [3H]thymidine incorporation (2.7-fold). Pressure-induced DNA synthesis was significantly suppressed by inhibitors of phospholipase C (2-nitro-4-carboxyphenyl-N,N-diphenylcarbamate), protein kinase C [1-(5-isoquinolinylsulfonyl)-2-methylpiperazine and chelerythrine], or tyrosine kinases (genistein). Pressure caused a rapid but transient formation of inositol 1,4,5-trisphosphate, which was blocked by the phospholipase C inhibitor. Pressure also promoted a rapid increase in tyrosine kinase activity. Pressure increased mRNA levels of PDGF-B, with a peak at 6 h, but not those of PDGF-A or bFGF. Pressure-induced DNA synthesis was partially inhibited by a neutralizing anti-PDGF antibody but not by an antibody against bFGF or nonimmune IgG. Our results indicated that pressure by itself increases DNA synthesis and proliferation of cultured rat MC possibly through activation of protein kinase C and tyrosine kinases, and PDGF-B could be partially involved in these pathways.

The crystal structure of tobacco PR-5d, an antifungal thaumatin-like protein isolated from cultured tobacco cells, was determined at the resolution of 1.8 Å. The structure consists of 208 amino acid residues and 89 water molecules with a crystallographic R-factor of 0.169. The model has good stereochemistry, with respective root-mean-square deviations from the ideal values for bond and angle distances of 0.007 Å and 1.542 °. Of the homologous PR-5 proteins, only those with antifungal activity had a common motif, a negatively charged surface cleft. This cleft is at the boundary between domains I and II, with a bottom part consisting of a three-stranded antiparallel β-sheet in domain I. The acidic residues located in the hollow of the cleft form the β-sheet region. Sequence and secondary structure analyses showed that the amino acid residues comprising the acidic cleft of PR-5d are conserved among other antifungal PR-5 proteins. This is the first report on the high-resolution crystal structure of an antifungal PR-5 protein. This structure provides insight into the function of pathogenesis-related proteins.

Phosphinic acid-, sulfoximine- and sulfone-based transition-state analogues were synthesized and evaluated as inhibitors of Escherichia coliγ-glutamylcysteine synthetase. These compounds have a carboxyl function at the β-carbon to the tetrahedral central hetero atom so as to mimic the carboxyl group of the attacking cysteine in the transition state. The phosphinic acid- and the sulfoximine-based compounds were found to be potent ATP-dependent inactivators, both showing a slow-binding kinetics with overall affinities and second-order inactivation rates of one to two orders of magnitude greater than those of l-buthionine (SR)-sulfoximine (l-BSO). The sulfone was a simple reversible inhibitor without causing ATP-dependent enzyme inactivation, but its affinity toward the enzyme was still five times greater than that of l-BSO, indicating that the β-carboxyl function plays a key role in the recognition of the inhibitors by the enzyme. The sulfoximine with (S)-β-carbon to the sulfur was synthesized stereoselectively, and the two diastereomers with respect to the chiral sulfur atom were separated as a cyclic sulfoximine derivative. The sulfoximine with R-configuration around the sulfur served as an extremely powerful ATP-dependent inactivator with an overall inhibition constant of 39nM and an inactivation rate of 6750M−1s−1, which correspond to 1260-fold higher affinity and almost 1400-fold greater inactivation rate as compared with l-BSO. The sulfoximine with (S)-sulfur was a simple reversible inhibitor with an inhibition potency comparable to that of the sulfone. The synthesis and inhibition profile of the N-phosphoryl sulfoximine is also described.

To determine whether human endogenous retroviruses are implicated in the pathogenesis of inflammatory vascular diseases of unknown etiology, we examined mRNA expression of a human endogenous retrovirus, HERV-R, which has a long open reading frame in the envregion, in cultured human vascular endothelial and smooth muscle cells stimulated in the presence of various cytokines. mRNA of HERV-R was always evident in these cells but not in fibroblastic cells. Levels of expression in vascular endothelial cells were significantly regulated by treatment with tumor necrosis factor-α, interleukin (IL)-1α, and IL-1β as up-regulators and interferon-γ as a down-regulator. These observations are interpreted to mean that HERV-R expression may be up- or down-regulated at sites of inflammation in vessels in vivoand hence may play a pathogenetic role in inflammatory vascular diseases in humans, perhaps similar to endogenous retroviruses in mouse models of polyarteritis nodosa in humans.

Adrenomedullin (AM), a hypotensive peptide isolated from human pheochromocytoma, inhibits the proliferation of mesangial cells (MC) induced by mitogens such as platelet-derived growth factor. Quite recently, we have demonstrated that transmural pressure applied to cultured MC increased DNA synthesis and cell proliferation through protein kinase C and tyrosine kinase pathways. However, the modulatory effect of AM on pressure-induced cell proliferation is as yet unknown. In the present study, we examined the effect of AM on transmural pressure-induced DNA synthesis in cultured rat MC. Pressure was applied to cells placed in a sealed chamber using compressed helium. Application of pressure resulted in an increase in [3H]thymidine incorporation (approximately 2.0-fold). AM clearly inhibited pressure-induced DNA synthesis in a concentration-dependent manner. This inhibition was paralleled by an increase in cellular cAMP levels evoked by AM. Forskolin and dibutyryl cAMP mimicked the inhibitory effect of AM. The protein kinase A inhibitor H-89 significantly attenuated the effect of AM. Human AM(22–52)-NH2, a putative AM receptor antagonist, reversed the inhibitory effects of AM more potently than did human CGRP(8–37), a calcitonin gene related peptide receptor antagonist. Our results suggest that AM, by acting mainly on AM-sensitive receptors, inhibits pressure-induced DNA synthesis in cultured rat MC through activation of protein kinase A. AM may play a protective role against MC proliferation in certain pathological conditions.

ABSTRACTUnlike ubiquitin, the ubiquitin-like protein modifier SUMO-1 and its budding yeast homologue Smt3p have been shown to be more important for posttranslational protein modification than for protein degradation. Here we describe the identification of the SUMO-1 homologue of fission yeast, which we show to be required for a number of nuclear events including the control of telomere length and chromosome segregation. A disruption of thepmt3+gene, the Schizosaccharomyces pombehomologue of SMT3, was not lethal, but mutant cells carrying the disrupted gene grew more slowly. Thepmt3? cells showed various phenotypes such as aberrant mitosis, sensitivity to various reagents, and high-frequency loss of minichromosomes. Interestingly, we found thatpmt3+is required for telomere length maintenance. Loss of Pmt3p function caused a striking increase in telomere length. When Pmt3p synthesis was restored, the telomeres became gradually shorter. This is the first demonstration of involvement of one of the Smt3p/SUMO-1 family proteins in telomere length maintenance. Fusion of Pmt3p to green fluorescent protein (GFP) showed that Pmt3p was predominantly localized as intense spots in the nucleus. One of the spots was shown to correspond to the spindle pole body (SPB). During prometaphase- and metaphase, the bright GFP signals at the SPB disappeared. These observations suggest that Pmt3p is required for kinetochore and/or SPB functions involved in chromosome segregation. The multiple functions of Pmt3p described here suggest that several nuclear proteins are regulated by Pmt3p conjugation.

Abstract: Magnetization and Mssbauer measurements have been made on a diluted antiferromagnets FexMg1−xTiO3 with x=0.2, 0.3 and 0.7. It has been demonstrated that this system exhibits a spin glass or a reentrant spin glass like behavior in the samples with x around the percolation concenrration xc≈0.25. It has been shown that various sizes of magnetic clusters are formed well above the transition temperatures determined by the magnetization measurements in the samples with x near xc. The behavior of Fe0.2Mg0.8TiO3 is discussed as a typical cluster glass.

The elution behavior of sulfated bile acids in high-performance liquid chromatography on the octadecylsilyl bonded column with acetonitrile/0.5% phosphate buffer has been investigated. A significant influence of pH of the mobile phase on the capacity ratio (κ') was observed in the higher pH region for bile acid 12-sulfates. Blockage of the 12α-hydroxyl group in sulfated bile acids by acetylation produced a marked decrease in the κ' values relative to their parent compounds in the pH range above 6.0. The κ' values of dehydrocholate monosulfates and their acetates were measured and the increments exerted by transformation into the acetates were estimated. Remarkable increments were observed for dehydrocholate monosulfates with the 3α-hydroxyl group but not for those with the 7α- or 12α-hydroxyl group. The effect of pH of the mobile phase on chromatographic behavior has been discussed from the stereochemical point of view.

The crystal structure of E. coli asparagine synthetase has been determined by X-ray diffraction analysis at 2.5 Å resolution. The overall structure of the enzyme is remarkably similar to that of the catalytic domain of yeast aspartyl-tRNA synthetase despite low sequence similarity. These enzymes have a common reaction mechanism that implies the formation of an aminoacyl-adenylate intermediate. The active site architecture and most of the catalytic residues are also conserved in both enzymes. These proteins have probably evolved from a common ancestor even though their sequence similarities are small. The functional and structural similarities of both enzymes suggest that new enzymatic activites would generally follow the recruitment of a protein catalyzing a similiar chemical reaction.

Two mutant forms, which had truncated N-terminals, of lipase activator protein (LipB) from Pseudomonas aeruginosaTE3285 were prepared, and their molecular properties and activity were compared with those of the full-length form. A truncated LipB lacking its hydrophobic N-terminal 21 residues was dispersed homogeneously in solution, and could reactivate the stoichiometric amount of denatured lipase. In contrast, full-length LipB formed soluble aggregates, and reactivated less than an equimolar amount of the lipase even under the most suitable conditions. These findings suggest that some or all of the N-terminal 21 residues caused aggregation of the protein molecules, and prevented LipB from fully stoichiometric reactivation. A truncated LipB lacking the N-terminal 61 residues also reactivated denatured lipase, suggesting that the N-terminal 61-residue region of LipB is not involved in reactivation.