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The ionome is defined as the mineral element composition of an organism or tissue and is genetically and environmentally influenced. Although nutrient deficiency changes the plant ionome, the differences among different species have not been elucidated fully. In the present study, we examined the ionomic responses to nutrient deficiency in wheat (Triticum aestivum L.), maize (Zea mays L.), sunflower (Helianthus annuus L.), and soybean (Glycine max (L.) Merr.) to understand the differences in response to nutrient deficiency among different plant species. The plants were cultivated in fields with four fertilizer treatments: complete fertilization, without nitrogen, without phosphorus, and without potassium. Each plant species was sampled just before the flowering stage, and mineral concentrations of each organ were analyzed. The species-specific and nonspecific changes in accumulation by the treatment were observed in different elements. Under phosphorus deficiency, sunflower showed a different variation in the ionome profile than the other species, particularly in the increased accumulation of many metal elements. Increased accumulation of many elements was observed in all plant species under potassium deficiency. Under nitrogen deficiency, leaf molybdenum accumulation was increased in non-leguminous plants. An unknown role of molybdenum in the metabolic responses to the nitrogen deficiency was predicted. To carry out further research based on the results of such ionomics studies may reveal the unknown function of essential elements in metabolic responses in plants.

Purpose We developed a system for calculating patient positional displacement between digital radiography images (DRs) and digitally reconstructed radiography images (DRRs) to reduce patient radiation exposure, minimize individual differences between radiological technologists in patient positioning, and decrease positioning time. The accuracy of this system at five sites was evaluated with clinical data from cancer patients. The dependence of calculation accuracy on the size of the region of interest (ROI) and initial position was evaluated for clinical use. Methods For a preliminary verification, treatment planning and positioning data from eight setup patterns using a head and neck phantom were evaluated. Following this, data from 50 patients with prostate, lung, head and neck, liver, or pancreatic cancer (n = 10 each) were evaluated. Root mean square errors (RMSEs) between the results calculated by our system and the reference positions were assessed. The reference positions were manually determined by two radiological technologists to best‐matching positions with orthogonal DRs and DRRs in six axial directions. The ROI size dependence was evaluated by comparing RMSEs for three different ROI sizes. Additionally, dependence on initial position parameters was evaluated by comparing RMSEs for four position patterns. Results For the phantom study, the average (± standard deviation) translation error was 0.17 ± 0.05, rotation error was 0.17 ± 0.07, and ΔD was 0.14 ± 0.05. Using the optimal ROI size for each patient site, all cases of prostate, lung, and head and neck cancer with initial position parameters of 10 mm or under were acceptable in our tolerance. However, only four liver cancer cases and three pancreatic cancer cases were acceptable, because of low‐reproducibility regions in the ROIs. Conclusion Our system has clinical practicality for prostate, lung, and head and neck cancer cases. Additionally, our findings suggest ROI size dependence in some cases.

At least 17 elements are known to be essential for plants; however, plants also absorb and accumulate various nonessential elements. Plants re-translocate different elements, including essential and nonessential elements, with differing efficiencies from mature to young developing organs via the phloem transport, resulting in different distributions of minerals in these organs. Therefore, in this study, we analyzed the concentrations of 23 elements in young and mature leaves of different leaf vegetable crops to comprehensively understand the mineral dynamics in plants. The allocation profiles of minerals in leaves of varying ages differed among species. For example, a higher molybdenum allocation profile was observed in young crown daisy ( Glebionis coronaria ) leaves, which might be related to the efficient nitrate assimilation in young leaves of this species. Thus, this study provides a new insight into the mineral uptake and transport mechanisms in plants.

Highly accurate radiotherapy needs highly accurate patient positioning. At our facility, patient positioning is manually performed by radiology technicians. After the positioning, positioning error is measured by manually comparing some positions on a digital radiography image (DR) to the corresponding positions on a digitally reconstructed radiography image (DRR). This method is prone to error and can be time‐consuming because of its manual nature. Therefore, we propose an automated measuring method for positioning error to improve patient throughput and achieve higher reliability. The error between a position on the DR and a position on the DRR was calculated to determine the best matched position using the block‐matching method. The zero‐mean normalized cross‐correlation was used as our evaluation function, and the Gaussian weight function was used to increase importance as the pixel position approached the isocenter. The accuracy of the calculation method was evaluated using pelvic phantom images, and the method's effectiveness was evaluated on images of prostate cancer patients before the positioning, comparing them with the results of radiology technicians' measurements. The root mean square error (RMSE) of the calculation method for the pelvic phantom was 0.23±0.05 mm. The coefficients between the calculation method and the measurement results of the technicians were 0.989 for the phantom images and 0.980 for the patient images. The RMSE of the total evaluation results of positioning for prostate cancer patients using the calculation method was 0.32±0.18 mm. Using the proposed method, we successfully measured residual positioning errors. The accuracy and effectiveness of the method was evaluated for pelvic phantom images and images of prostate cancer patients. In the future, positioning for cancer patients at other sites will be evaluated using the calculation method. Consequently, we expect an improvement in treatment throughput for these other sites. PACS number: 87

The purpose of this study is development of Mobile Gestural Control System of a Vehicle. The designated coordinate was estimated from the intersection of the location information and the floor of the elbow and wrist. Position control system for moving the vehicle indicated coordinate was designed. When there is an obstacle on the path of the robot, vehicle avoid the obstacle by potential method.

The purpose of this paper is an estimation of driving torque by using an external sensor. This paper deals with an estimation method of driving torque and physical parameters. This research have been estimated the driving torque of each joints as the force. Future, this research will design an estimation system of a myoelectric signal using an external sensor. Moreover, we aim to design an evaluation system of the ability to exercise using obtained biosignal.

Background/aim To analyze the accuracy of patient positioning and dose distribution quality using a fiducial marker-matching technique in carbon-ion radiotherapy (C-ion RT) for stage I lung cancer. Patients and methods Thirteen patients with 26 fiducial markers and 104 radiation fields were examined. Change in the fiducial marker position coordinates from the gross tumor volume center (Δm), and change in the water-equivalent path length of the chest wall (ΔWEL) were measured in each radiation field. Criterion for an acceptable change in dose distribution was defined as the percentage of D95 (%D95) at gross tumor volume greater than 90% of the prescribed dose. Results Thirteen radiation fields (12.5%) were classified as unacceptable. Δm and ΔWEL had significant negative correlations with %D95 and thus were significant factors related to unacceptable irradiation fields. Conclusion Patient positioning using a fiducial marker-matching technique resulted in 12.5% of radiation fields demonstrating unacceptable degradation due to Δm and ΔWEL.

Background and purpose Patient positioning was compared by tumor matching (TM) and conventional bony structure matching (BM) in carbon ion radiotherapy for stage I non-small cell lung cancer to evaluate the robustness of TM and BM in determining interfractional error. Material and methods Sixty irradiation fields were analyzed. Computed tomography (CT) images acquired before treatment initiation for confirmation (Conf-CT) were obtained under the same settings as the treatment planning CT images and used to evaluate both positioning methods. The dose distributions were recalculated for Conf-CT using both BM and TM, and the dose–volume histogram parameters [V 95% of clinical target volume, V 5Gy(RBE) of normal lung, and acceptance ratio (ratio of cases with V 95% > 95%)] were evaluated. The required margin, which in 90% of cases achieved the acceptable condition, was also examined. Results Using BM and TM, the median V 95% was 98.93% and 100% ( p 5Gy(RBE) was 135.9 and 125.8 ( p = 0.694), respectively. The estimated required margins were 7.9 and 3.3 mm and increased by 53.9% and 2.5% of V 5Gy(RBE) , respectively, compared with planning. Conclusions TM ensured a better dose distribution than did BM. To enable TM, volumetric imaging is crucial and should replace 2D radiographs for carbon therapy of stage I lung cancer.

Purpose The contrast agent used in the diagnostic department has high atomic numbers and might influence dose deposition in the particle therapy. In particular, the influence of gadolinium-based (Gd) contrast agent on range in carbon ion radiotherapy has not yet been evaluated. For this reason, we avoid carbon treatment and planning computed tomography (CT) acquisition on days when the contrast-enhanced magnetic resonance image (MRI) is performed. In this study, we evaluated the time required for this beam range effect to vanish by evaluating the temporal changes in the CT values after an enhanced MRI as well as the stopping power of Gd solution. Materials and methods Two types of diluted solutions with Gd contrast agent were used for comparing their transferred stopping power (TSP) and measured stopping power (MSP). The TSP was calculated with a CT value to stopping power ratio table that was created previously. Additionally, to evaluate in vivo attenuation, we measured the CT values in the renal pelvis from the CT images with and without contrast agent for 73 patients. Results The maximum difference between the TSP and MSP was 85%. The difference between the TSP after 4 hours and the TSP with non-enhanced cases was less than 1%. Moreover, the difference between the MSP after 1 hour and the MSP with non-enhanced cases was less than 0.1%. Conclusion It was found that the impact of Gd contrast agent can be neglected 1 hour after administration for carbon beam irradiation and 4 hours after for planning the CT image acquisition.

PURPOSE We developed an evaluation method for easily calculating displacement directly between the carbon beam axis and positioning X-ray axis. METHODS A verification image was acquired by irradiating an imaging plate with a carbon beam and X-ray. The X-ray passed through a lead plate inserted in the range compensator holder. The displacement was calculated on the verification image from the center of a wire irradiated with carbon using a multi leaf collimator (MLC) and a wire irradiated with X-ray also using MLC. The accuracy of the method was evaluated by moving the carbon beam axis, the X-ray axis, and the setup angle. The weekly changes of vertical and lateral beams in all rooms were also evaluated. RESULTS The displacements of the carbon beam axis and the setup angle did not influence the calculation results, whereas the displacement of the X-ray axis did (R=0.999). The displacements including weekly changes were all less than 1.00 mm. CONCLUSION An evaluation method for calculating the displacement directly and simply between the carbon beam axis and positioning X-ray axis was developed and verified. The weekly changes of displacement between axes were evaluated to be acceptable at our facility.

The superconducting transition temperatures (TC) of (1 1 1) and (0 0 1) boron-doped diamond films deposited by microwave plasma assisted chemical vapor deposition (MPCVD) are investigated in the wide boron concentration range of 1 × 1020 − 1 × 1022 cm−3. The critical boron concentrations of superconductor to insulator transition in (1 1 1) and (0 0 1) films are 3 × 1020 cm−3. TC in (1 1 1) films does not have the tendency to saturate up to 1 × 1022 cm−3, while TC in (0 0 1) films saturates.

We investigated the diagnostic significance of IL-6 for lymph node metastasis and/or hepatic metastasis from colorectal cancer in 65 patients and evaluated the contributions of 8 factors (IL-6, HGF, IL-1beta, TNF-alpha, TGF-beta1, ELAM-1, ICAM-1, VCAM-1) toward Dukes.s classification of 53 patients. We also examined IL-6 expression in tumor tissue. From the receiver operating characteristic (ROC) curve analysis, an optimal cutoff value of 5.8 pg/ml was determined to classify lymph node and/or hepatic metastasis, and that of 6.3 pg/ml was determined to classify hepatic metastasis. These values indicated sensitivities of 55.0% and 71.4%, and specifi cities of 100% and 88.6%, respectively. IL-6, HGF, and ELAM-1 were very useful for distinguishing among Dukes.s A/B group, C group, and D group. In all cases with high IL-6 values (more than 25.0 pg/ml), immunohistochemical staining was positive for IL-6 in the cytoplasm of cancer cells. IL-6 is strongly suspected to be involved in lymph node and/or hepatic metastasis by promoting it through HGF, and serum IL-6 value (pg/ml) would be useful diagnostically to estimate whether or not there is a high risk of lymph node and/or hepatic metastasis.

症例は42歳の男性で, 胸部中部進行食道癌に対し術前化学放射線療法を施行後, 右開胸開腹食道亜全摘, 3領域リンパ節郭清 (胸管温存) を行った. 第3病日より胸腔ドレーンの排液が急増し, 翌日経腸栄養を開始したところ排液が白濁したため乳糜胸と診断した. 同日総排液量は1,800mlに達し循環動態に影響が出たため手術を施行した. 下縦隔で胸管をクリッピングしたが上縦隔からの乳糜漏出は停止せず, 上縦隔左側を展開し胸管の乳糜漏出部を確認し, その足側をクリッピングして漏出を停止できた. しかし, 翌日にはドレーンから再度多量の乳糜流出が認められたため, オクトレオチド50μg×3回/日の間歇皮下投与を試みた. 2日目から排液は著明に減少し, 計8日間漸減投与し中止としたが乳糜漏の停止が確認され退院できた. 食道癌術後の乳糜胸には難渋することがあるが, オクトレオチド間歇皮下投与は, 副作用も認められず試みる価値があるものと思われた.

腫瘍はさまざまなサイトカインを産生し,その進展を促すとともに随伴症状を呈する.今回,胃癌術後類白血病反応を呈したG-CSF, IL-6を産生する再発例を経験した.症例は55歳,男性.平成13年12月胃全摘術が施行され,翌年11月より39°C台の発熱, WBC, CRP, CEA, CA19-9などの著明な上昇を認めた.骨髄検査では,反応性の骨髄過形成以外異常所見はみられず血液疾患は否定された.左腎上極外側に6×4cmの腫瘤を認め,組織学的に胃癌の再発腫瘍と診断された.また,腫瘍の細胞質にG-CSF, IL-6の陽性所見を認め各血清値 (242.0pg/ml, 60.3pg/ml) への反映が示唆された.病状はしだいに進行し類白血病反応を呈するとともに,癌性悪液質のため平成15年1月永眠された.死亡前の白血球数114,960/μl, CRP 16.18mg/dl, CEA 229.79ng/ml, CA19-9 491.57U/mlまで上昇した.術後の急速な進展と生体反応は, G-CSFおよびIL-6の強い影響が示唆された.

Objective: We compared the efficacy of plastic prostheses and self-expandable metallic stents in the treatment of malignant esophageal stenosis and/or fistula.Subjects and Methods: Subjects were 31 patients with esophageal cancer, 4 with esophagotracheal fistula, and 1 with esophageal stenosis. A plastic prosthesis was inserted in 15 patients (group A) and a self-expandable metallic stent in 21 patients (group B). We evaluated food ingestion improvement, effectiveness, complications, mean survival, food ingestion duration, the percentage of food ingestion capability in total survival time, and inhospital mortality.Results: No differences were seen in food intake improvement or in the effectiveness between groups, whereas fatal complications were higher in Group A. No significant differences were seen in mean survival, food ingestion duration, percentage of food ingestion capability, or inhospital mortality between groups.Conclusion: We concluded that a self-expandable metallic stent was safer than a plastic prosthesis because of fewer serious complications such as bleeding, and recommended the use of metallic stents in the treatment of malignant esophageal stenosis and/or fistula.

A method for hand-assisted laparoscopic construction of gastric conduit for thoracic esophageal cancer was developed. Since this endoscopic surgical procedure is less invasive than open surgery, it contributes to improvement of post-operative respiratory functions and reduces respiratory complications. What distinguishes our surgical procedure is that unlike methods described in previous reports, it begins with treatment of the left gastroepiploic vessels at the height of the inferior edge of the spleen, followed by dissection from the esophageal hiatus to the lesser curvature and then dissection and excision of left gastric arteries and veins. Finally, the exposed esophagus and stomach are drawn outside the body and the right gastroepiploic blood vessels are preserved, followed by dissection of the greater omentum. This approach to gastric conduit construction was undertaken in 6 patients and the mean operating time was 123 minutes. Although in the first 3 of these patients the operating time was 150 minutes or more, the time required shortened to around 90 minutes for each of the last 3 cases, as the procedure was mastered. In each case, the volume of intraoperative hemorrhage did not exceed 50 ml.

The film thickness dependence of Tc in heavily B-doped diamond (1 1 1) and (0 0 1) films was investigated and cross-sectional transmission electron microscope observation was employed to investigate the crystalline structure of heavily B-doped diamond. (1 1 1) films with 5 nm or more and (0 0 1) films with 40 nm or more show superconducting transition. Few dislocations are observed in 500 nm from the interface between the substrate and B-doped homoepitaxial layer in (1 1 1) film and defective structure observed in (0 0 1) film.

We encountered 588 patients with esophageal carcinoma between 1986 and 1998. For 96 (16.4%) of the patients 75-year-old or older, we examined the features and issues of esophageal carcinoma in elderly people based on preoperative conditions and treatment results. 1) Cardiac dysfunction was observed in 39 patients (40.6%). Concerning respiratory function, there was a significant difference in forced expiratory volume in 1 second (FEV1.0%) and % vital capacity (%VC) compared to patients under 75 years of ages (t-test). Furthermore, disorders of several organs were observed in 31 patients (32.2%). 2) Eleven patients underwent right thoracotomy and laparotomy. Three (25%) of these patients died in hospital postoperatively. The 3-year survival rate was 27.3%. 3) Fourteen patients underwent esophagectomy. Four (28.6%) of these patients died in hospital. The 3-year survival rate was 7.7%. 4) In 66 patients who did not undergo resection, the 3-year survival rate was 11.7%. 5) With respect to postoperative complications, the incidences of pulmonary complications and postoperative delirium were high. Some patients died during surgery or during hospitalization. Careful perioperative management is important. 6) There was no significant difference in long-term results between treatment methods. However, none of the patients who underwent C0 surgery survived more than 2 years. Therefore, this procedure should be avoided if possible.

従来より食道気管支瘻に対して, 保存的治療, 内視鏡治療, 手術などが行われてきた. 今回, 我々は生体接着剤であるEthyl-2-cyanoacrylate (以下, ECAと略す) を用いた内視鏡的瘻孔閉鎖術を試み, 良好な結果を得たので報告する.症例は73歳の男性. 平成9年6月初めより胸部不快感出現し, 6月6日内視鏡検査にて上切歯列より32cm, 3時方向に憩室があり, 憩室内に2mmの瘻孔を認めた. 7月2日瘻孔造影にて右B6から造影剤の流出を認めた. 7月10日散布チューブよりECA 1mlを瘻孔開口部に散布し, 造影にて瘻孔が閉鎖しているのを確認した. 散布後4日目の造影で一部造影剤の流出を認めたため, 再度ECA 1mlを散布した. 翌日より食事を開始し退院となった. ECAによる内視鏡的瘻孔閉鎖術は, 比較的簡単な手技で安全に繰り返し施行できるため, 手術を考慮する前や, 全身状態の悪い症例に有効な方法と思われた.

Soil contains various essential and nonessential elements, all of which can be absorbed by plants. Plant ionomics is the study of the accumulation of these elements (the ionome) in plants. The ionomic profile of a plant is affected by various factors, including species, variety, organ, and environment. In this study, we cultivated various vegetable crop species and cultivars under the same field conditions and analyzed the level of accumulation of each element in the edible and nonedible parts using ionomic techniques. The concentration of each element in the edible parts differed between species, which could be partly explained by differences in the types of edible organs (root, leaf, seed, and fruit). For example, the calcium concentration was lower in seeds and fruit than in other organs because of the higher dependency of calcium accumulation on xylem transfer. The concentration of several essential microelements and nonessential elements in the edible parts also varied greatly between cultivars of the same species, knowledge of which will help in the breeding of vegetables that are biofortified or contain lower concentrations of toxic elements. Comparison of the ionomes of the fruit and leaves of tomato (Solanum lycopersicum) and eggplant (S. melongena) indicated that cadmium and boron had higher levels of accumulation in eggplant fruit, likely because of their effective transport in the phloem. We also found that homologous elements that have been reported to share the same uptake/transport system often showed significant correlation only in a few families and that the slopes of these relationships differed between families. Therefore, these differences in the characteristics of mineral accumulation are likely to affect the ionomic profiles of different families.

We propose a strategy of individualized image acquisitions and treatment planning for respiratory-gated carbon-ion therapy. We implemented it in clinical treatments for diseases of mobile organs such as lung cancers at the Gunma University Heavy Ion Medical Center in June 2010. Gated computed tomography (CT) scans were used for treatment planning, and four-dimensional (4D) CT scans were used to evaluate motion errors within the gating window to help define the internal margins (IMs) and planning target volume for each patient. The smearing technique or internal gross tumor volume (IGTV = GTV + IM), where the stopping power ratio was replaced with the tumor value, was used for range compensation of moving targets. Dose distributions were obtained using the gated CT images for the treatment plans. The influence of respiratory motion on the dose distribution was verified with the planned beam settings using 4D CT images at some phases within the gating window before the adoption of the plan. A total of 14 lung cancer patients were treated in the first year. The planned margins with the proposed method were verified with clinical X-ray set-up images by deriving setup and internal motion errors. The planned margins were considered to be reasonable compared with the errors, except for large errors observed in some cases.

The critical concentration of superconductor-to-insulator transition in boron-doped diamond is determined in two ways, namely, the actual doping concentration of boron and the Hall carrier concentration. Hall carrier concentrations in (111) and (001) films exceed the actual doping concentration owing to the distortion of the Fermi surface. The high critical boron concentration in (110) films is owing to the effect of the high concentration of interstitial boron atoms. A boron concentration of $3\ifmmode\times\else\texttimes\fi{}{10}^{20}\text{ }{\text{cm}}^{\ensuremath{-}3}$ in substitutional site is required for inducing superconductivity in diamond.

Ionomics is the study of elemental accumulation in living organisms using high-throughput elemental profiling. In the present study, we examined the ionomic responses to nutrient deficiency in maize grown in the field in long-term fertilizer trials. Furthermore, the available elements in the field soils were analyzed to investigate their changes under long-term fertilizer treatment and the ionomic relationships between plant and soil. Maize was cultivated in a field with the following five long-term fertilizer treatments: complete fertilization, fertilization without nitrogen, without phosphorus, without potassium, and no fertilization. Concentrations of 22 elements in leaves at an early flowering stage and in soils after harvest were determined. The fertilizer treatments changed the availabilities of many elements in soils. For example, available cesium was decreased by 39 % and increased by 126 % by fertilizations without nitrogen and potassium, respectively. Effects of treatments on the ionome in leaves were evaluated using the translocation ratio (the concentration in leaves relative to the available concentration in soils) for each element. Nitrogen deficiency specifically increased the uptake ability of molybdenum, which might induce the enhancement of nitrogen assimilation and/or endophytic nitrogen fixation in plant. Potassium deficiency drastically enhanced the uptake ability of various cationic elements. These elements might act as alternatives to K in osmoregulation and counterion of organic/inorganic anions. Two major groups of elements were detected by multivariate analyses of plant ionome. Elements in the same group may be linked more or less in uptake and/or translocation systems. No significant correlation between plant and soil was found in concentrations of many elements, even though various soil extraction methods were applied, implying that the interactions between the target and other elements in soil must be considered when analyzing mineral dynamics between plant and soil. Electronic supplementary material The online version of this article (doi:10.1186/s40064-015-1562-x) contains supplementary material, which is available to authorized users.

Background. It is becoming clear that various cytokines are associated with the spread of cancer cells. The purpose of this study was to compare interleukin (IL)-6 levels in patients with gastric cancer to elucidate the role of IL-6 in predicting the spread of tumors. Methods. In 60 patients, we assessed the correlation of serum IL-6 (pg/ml) with stage, histological findings, hepatic metastasis, and related factors (hepatocyte growth factor [HGF], IL1� , tumor necrosis factor [TNF]-α, and transforming growth factor [TGF]-� 1). We also investigated the diagnostic significance of the IL-6 level for advanced gastric cancer and lymph node metastasis, as well as the association between IL-6 elevation and outcome. Finally, we examined the expression of IL-6 in tumor tissue. Results. Significant relationships were seen between serum IL-6 and stage, depth of tumor invasion (pT), lymphatic invasion (ly), venons invasion (v)*, lymph node metastasis (pN), hepatic metastasis (cH), and HGF (P 0.01; *P 0.05). With regard to the diagnostic significance of the IL-6 level for advanced gastric cancer and lymph node metastasis, when the cutoff value of IL-6 was set at 1.97 pg/ml, the sensitivity was 81.8% and 87.5%; specificity was 66.7% and 58.3%; and accuracy was 77.1% and 72.9%, respectively. The 1- and 3-year cumulative survival rates for patients with an IL-6 value of more than 1.97 pg/ml (69.0% and 43.4%, respectively) were significantly lower than those for patients with an IL-6 value of 1.97 pg/ml or less (94.4% and 87.2%, respectively; P 0.05). Immunohistochemical staining was positive for IL-6 in the cytoplasm of cancer cells. Conclusion. We suspect that IL-6 is involved in cancer invasion and lymph node and/or hepatic metastasis. Our results indicate that IL-6 could be used as a prognostic factor for survival.

The homolog of the dnaA gene, which has been reported to be present in the vicinity of the initiation site of replication in the genome of Mycoplasma capricolum (M.Miyata, L.Wang, and T.Fukumura, J. Bacteriol. 175: 655-660, 1993) was mapped precisely. A 9540-bp region containing the dnaA gene was cloned and the entire region was sequenced with the exception of a previously reported region of 2517 bp (Fujita, M.Q., Yoshikawa, H. and Ogasawara, N. Gene 93: 73-78, 1992). The organization of the 9540-bp region was compared with that of corresponding regions in other bacteria. The arrangement and directions of rnpA, rpmH, dnaA, dnaN were conserved, but no other open reading frames were found that were homologous to those that are commonly found around dnaA genes in other bacteria. The directions of movement of the replication fork around the dnaA gene were analyzed by neutral/alkaline two-dimensional gel electrophoresis. The forks developed in a 1569-bp region that consisted of the dnaA structural gene and its downstream non-coding region, and then they proceeded bidirectionally.