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The structure of the isolated alpha subunit of F1-ATPase from the thermophilic Bacillus strain PS3 was probed using limited proteolysis by four different proteases, and the following results were obtained. 1) Distribution of 21 protease-cleaved sites is similar to that of the beta subunit of F1-ATPase (Tozawa, K., Odaka, M., Date, T., and Yoshida, M. (1992) J. Biol. Chem. 267, 16484-16490), thus providing experimental evidence for similar folding topology of the two subunits, and the locations of 11 water-exposed loop regions in the tertiary structure are predicted. 2) Most proteolytic peptides remain associated to maintain the gross structure of the alpha subunit and can reassociate each other after denaturing urea treatment. 3) However, the carboxyl-terminal peptides comprising approximately 80 residues (C1 peptides) are released from other peptide(s) during proteolysis, and those comprising approximately 105 residues (C2 peptides) are released during native polyacrylamide gel electrophoresis after proteolysis. 4) Inclusion of Mg-ATP in the native electrophoretic system prevents the release of the C2 peptide. Addition of Mg-ATP to the proteolysis mixtures results in an increase of the C2 peptide population and a decrease of the C1 peptide population. Thus, Mg-ATP induces a conformational change at the regions of C1 and C2 peptides of the alpha subunit. 5) Except for the trypsin-treated one, protease-treated alpha subunits are reconstitutable with the native beta subunit into the form of alpha 3 beta 3 complexes, which show significantly higher ATPase activities than the intact alpha 3 beta 3 complex. This activation is attributable to the cleavage of a peptide bond that produces C2 peptides. The carboxyl-terminal region of the alpha subunit is likely to be involved in the regulation of ATPase activity in F1-ATPase.

Partial digestion of the native beta subunit of F1-ATPase from the thermophilic Bacillus strain PS3 by three different proteases produced a limited number of peptide fragments. In most cases, the peptides remained associated, and the gross structure of the beta subunit was not destroyed. Furthermore, most peptides were able to reassociate into the form of the beta subunit after denaturating urea treatment. Therefore, the cleaved sites are most likely located in water-exposed loop regions in the tertiary structure of the protein. Almost all peptides were analyzed, and 17 cleaved sites were determined. From the analysis of the distribution of cleaved sites and deletions or insertions in the multiple amino acid sequence alignment of proteins homologous to the beta subunit, locations of five loops and four candidate loops in the beta subunit are suggested. There are two large loops in the central region of the beta subunit sequence, and dicyclohexylcarbodiimide-reactive Glu190 is located in one of them. Tyr341, involved in putative catalytic ATP binding, is also found in one of the loops. Then, taking cleaved sites as a reference, two kinds of expression plasmids, each of which carried genes of two complementary peptide fragments, 1-193 and 198-473 or 1-284 and 285-473, were constructed and expressed in Escherichia coli. For each plasmid, two peptides were coexpressed, associated into a stable beta subunit form in E. coli cells, and purified without dissociation. When these beta subunits were denatured by urea and applied to polyacrylamide gel without denaturant, a protein band with the same mobility as that of the beta subunit appeared, indicating that reassociation of peptide fragments into the form of the beta subunit occurred upon removal of urea. These beta subunits retained the ability to reconstitute the alpha 3 beta 3 gamma complexes even though the efficiency of reconstitution and the recovered ATPase activities were decreased. These complexes were stable at high or low temperature, and ATPase activities were sensitive to inhibition by N3-.

More than 75% of zinc metal in the world is produced by the roast-leach-electrowinning process. In the roast-leach process, very pure zinc sulphate solution must be prepared for zinc electrowinning, because the zinc ion is less noble than the hydrogen ion. Therefore, the purification process is one of the most important unit processes in zinc hydrometallurgy. In general, the cobalt ion is focused on as a typical impurity in zinc electrowinning and several purification processes for cobalt removal have been adopted such as the beta-naphthol process, and the arsenic trioxide or antimony trioxide process. Recently, modernized plants are using the arsenic trioxide or antimony trioxide process, but there are very few published papers on the chemistry of these purification processes. In this paper, firstly the potential-pH diagrams for the systems MAsH2O and MSbH2O are established for conditions which are close to the operating conditions, and the chemistry of the purification process is deduced from the potential-pH diagrams. Secondly, the phenomena influenced by the chemistry are confirmed experimentally and then a comparison between arsenic and antimony trioxide purification is made based on the experimental results.

Summary Regulatory T cells (Treg) have an essential role in maintaining immune tolerance in the gut. The functional CD4+ Treg express the transcription factor forkhead box protein 3 (FoxP3) or a CD25high in humans. Further, depletion of elevated granulocytes/monocytes by extracorporeal adsorption (GMA) induces immunomodulation in patients with ulcerative colitis (UC). We investigated the impact of GMA on Treg. Thirty-one UC patients, clinical activity index (CAI) 12·1 ± 2·97, refractory to conventional medications including intravenous corticosteroid and 13 healthy controls (HC), were included. Patients received five GMA sessions over 5 weeks. Biopsies from the rectal mucosa and blood samples at baseline and post-GMA were immunostained with anti-CD4/FoxP3 and anti-CD4/CD25 antibodies for immunohistochemistry and flow cytometry. Following GMA, 22 of 31 patients achieved remission (CAI ≤ 4, P

This paper reports that the countermeasures against X‐ray intensity fluctuation have been developed on BL11XU at SPring‐8. Observed X‐ray intensity fluctuation might be caused from mechanical vibrations of monochromator crystals. So, we measured the vibrations under various conditions to identify vibration sources and took countermeasures against the mechanical vibrations. As a result, the methods successfully removed the mechanical vibrations and the X‐ray intensity fluctuation was reduced to about half.

A new JAEA beamline with an in‐vacuum undulator has been constructed at BL22XU in SPring‐8. The wide band energy X‐rays facilitate crystal structure studies under pressure, resonant X‐ray scattering experiments, X‐ray absorption fine structure as well as stress measurements. The energy range from 3 to 70keV with a high flux of X‐rays has been achieved at the beamline, using two sets of double crystal monochromators equipped with liquid nitrogen cooling Si (111) crystals. The photon flux at 3, 9, 25 and 70keV with ΔE/E ≒ 10−4 are about 2 × 1010, 3 × 1013, 5 × 1012 and 5 × 1010 photons/sec, respectively. Fluctuations in X‐ray intensities and position caused by the liquid nitrogen cooling system have been prevented by using MOSTAB, a feedback control system.

In situ fracture studies have been carried out on thin films of the NiTi intermetallic compound under plane stress, tensile loading conditions in the high-voltage electron microscope. Local stress-induced amorphization of regions directly in front of moving crack tips has been observed. The upper cutoff temperature, TC–Amax, for the stress-induced crystalline-to-amorphous transformation was found to be 600 K, identical to that for heavy ion-induced amorphization of NiTi and for ion-beam mixing-induced amorphization of Ni and Ti multilayer specimens. 600 K is also both the lower cutoff temperature, TA–Cmin, for radiation-induced crystallization of initially-unrelaxed amorphous NiTi and the lowest isothermal annealing temperature, TXmin, at which stress-induced amorphous NiTi crystallizes. Since TXmin should be TK, the ideal glass transition temperature, the discovery that TC–Amax=TA–Cmin=TXmin=TK implies that disorder-driven crystalline-to-amorphous transformations result in the formation of the ideal glas...

Liquid‐N2‐cooled Si crystals replaced water‐cooled diamond crystals for the double‐crystal monochromator on the JAERI beamline, BL11XU, at SPring‐8. To use the whole energy from 6 to 70 keV, we adopted the system of dual symmetric Si geometry, Si(111)/Si(311), which can be switched by a horizontal translation (x) stage perpendicular to the beam axis. The characterization of the crystals was performed by the use of both the x‐ray generator and the synchrotron radiation from the undulator. The observed intensities and rocking curves resulted in that each crystal was good for BL11XU and that the performance of x‐ray reflection was highly improved.

Flow-induced vibrations occur in general heat exchangers and nuclear reactors. They cause the tube failures through fretting wear or fatigue. Fluid elastic instability may lead to vibration amplitudes large enough to cause tube-to-tube clashing and in such cases will lead to relatively rapid failures. Therefore mechanisms of elastic vibration induced by cross flow are significant problems for large vibration amplitudes. The aims of this study are to analyze the vibration of cylinders at various flow velocities and to get the fundamental data of design for the prevention of accidents in heat exchanger. In this study, the experimental apparatus was built so that cylinder tip motions could be measured at various flow velocities. The experiments were conducted in different cylinder arrays. The cylinder vibration was induced in normal direction to flow by an alternate vortex. Moreover, once the flow velocity was increased to a certain value, the cylinder oscillation amplitude increased rapidly with flow and had a maximum. When five cylinders were arranged in a row normal to flow, the wandering of energy was generated between the two cylinders. For three cylinders arranged in tandem, two cylinders in upstream and downstream positions mutually vibrated out of phase. For many cylinders, the entropy had the maximum at a specific flow velocity and oscillating behavior of cylinders became chaotic state which could not predict its behavior.Copyright © 2002 by ASME

The C2 proton signals of all (twelve) histidine residues of the TF1 beta subunit in the 1H-NMR spectrum have been identified and assigned by means of pH change experiments and site-directed substitution of histidines by glutamines. pH and ligand titration experiments were carried out for these signals. Furthermore, the ATPase activity of the reconstituted alpha3beta3gamma complex was examined for the twelve mutant beta subunits. Two of three conserved histidines, namely, His-119 and 324, were found to be important for expression of the ATPase activity. The former fixes the N-terminal domain to the central domain. His-324 is involved in the formation of the interface essential for the alpha3beta3gamma complex assembly. The other conserved residue, His-363, showed a very low pK(a), suggesting that it is involved in the tertiary structure formation. On the binding of a nucleotide, only the signals of His-173, 179, 200, and 324 shifted. These histidines are located in the hinge region, and its proximity, of the beta subunit. This observation provided further support for the conformational change of the beta monomer from the open to the closed form on the binding of a nucleotide proposed by us [Yagi et al. (1999) Biophys. J. 77, 2175-2183]. This conformational change should be one of the essential driving forces in the rotation of the alpha3beta3gamma complex.

The antigenic P64K protein from the pathogenic bacterium Neisseria meningitidis is found in the outer membrane of the cell, and consists of two parts: an 81-residue N-terminal region and a 482-residue C-terminal region. The amino-acid sequence of the N-terminal region is homologous with the lipoyl domains of the dihydrolipoyl acyltransferase (E2) components, and that of the C-terminal region with the dihydrolipoyl dehydrogenase (E3) components, of 2-oxo acid dehydrogenase multienzyme complexes. The two parts are separated by a long linker region, similar to the linker regions in the E2 chains of 2-oxo acid dehydrogenase complexes, and it is likely this region is conformationally flexible. A subgene encoding the P64K lipoyl domain was created and over-expressed in Escherichia coli. The product was capable of post-translational modification by the lipoate protein ligase but not aberrant modification by the biotin protein ligase of E. coli. The solution structure of the apo-domain was determined by means of heteronuclear NMR spectroscopy and found to be a flattened beta barrel composed of two four-stranded antiparallel beta sheets. The lysine residue that becomes lipoylated is in an exposed beta turn that, from a [1H]-15N heteronuclear Overhauser effect experiment, appears to enjoy substantial local motion. This structure of a lipoyl domain derived from a dihydrolipoyl dehydrogenase resembles that of lipoyl domains normally found as part of the dihydrolipoyl acyltransferase component of 2-oxo acid dehydrogenase complexes and will assist in furthering the understanding of its function in a multienzyme complex and in the membrane-bound P64K protein itself.

A 56-year-old female underwent lobectomy with ND2a lymph node dissection for left lung cancer in April 1999. Histopathological examination demonstrated moderately differentiated adenocarcinoma (pT2N2M0, stage III A). She received one course of a combination of etoposide and cisplatin as adjuvant therapy, followed by oral intake of UFT. In November 1999, a left para-aortic lymph node recurrence was found. She received radiation therapy (total 60 Gy) to the mediastinum. In April 2000, new lung and left supraclavicular lymph node recurrences were found. She received three courses of vinorelbine 20 mg/m2 (days 1, 8) and cisplatin 80 mg/m2 (day 1) followed by radiation therapy (total 50.4 Gy) to the left supraclavicular lesion. After the chemotherapy, a complete response (CR) of all metastatic lesions was achieved. Adverse reactions were grade 1 alopecia, grade 2 nausea/vomiting, grade 3 neutropenia, hypochromia, and injection site reaction. The combination of vinorelbine and cisplatin is a useful regimen in non-small-cell lung cancer.

Summary The introduction of the multichannel autoanalyser made measurement of serum calcium concentrations easier, and led to a dramatic change in clinical presentations. The reliable methods such as computed tomography (CT), ultrasonography (US) and magnetic resonance imaging (MRI) for preoperative localization of abnormal parathyroid glands has long been sought to increase the cure rate of surgical treatment. We report the clinical feature of primary hyperparathyroism (PHPT). Patients were classified into four stages in chronological order. The early patients (the first stage 1970–1979) were mainly diagnosed in the treatment of urolithiasis. Approximately 20% of patients in the second stage (1980–1986) were symptom-free, and hypercalcemia was detected by autoanalyzer. Patients in the third stage (1987–1993) underwent preoperative localization studies including CT, scintigraphy, ultrasonography and MRI. The recent patients (the fourth stage, 1993–1999) were mostly treated in the present hospital. In the first stage, PHPT was an uncommon metabolic disorder that was typically associated with nephrolithiasis and was two to three times more common in men than in women. In the second, third and fourth stages. PHPT is a common and often symptomless endocrine disorder. The ratio of male to female is decreasing, because men are dominant in stone-formers. Four parathyroid glands were searched carefully in the first and second stages, and unilateral cervical exploration was performed in some preoperatively localized parathyroid glands in the third and fourth stages.

One of the delicate problems in a cancer treatment is the acquisition of a tolerance for a medicine. Antiandrogen treatment is valid in treatment of prostate cancer, but the disease develops into hormone-refractory prostate cancer after a uniformity period. The molecular mechanisms of recurrence and endocrine therapy failure in prostate cancer have remained unclear. In this paper we discuss the hormone-refractory prostate cancer that has been reported by many investigators.

A 58-year-old man was admitted with a swelling in the frontal region. Computerized tomography scan, magnetic resonance imaging and angiography revealed a tumor in the right frontal sinus. The surgically extirpated specimen showed clear cell carcinoma which was suspected to be a metastasis from renal cell carcinoma. Subsequent urologic examination disclosed the right renal tumor. Since there were no other systemic metastases, right nephrectomy was performed. Pathologically, the renal tumor was clear cell subtype renal cell carcinoma and had the same histology as the frontal sinus tumor.

A 41-year-old man was admitted to our hospital with chief complaints of painless tumor in the left scrotal contents and loss of body weight. As the tumor was suspected of being a neoplastic lesion, left radical inguinal orchiectomy was carried out. Histopathological examination showed metastatic adenocarcinoma of the bile duct. Metastatic carcinoma to the testicle, epididymis, and their tunics is unusual and testicular metastases from adenocarcinoma of the bile duct is extremely rare. A case of testicular metastases from carcinoma of the bile duct is reported and the literature reviewed.

Sairei-to has been reported to inhibit granulation and fibroblast proliferation. We administered Sairei-to (9.0 g/day) to two 77-year-old men with repeated urethral stenosis after transurethral resection of prostate (TUR-P) and examined its clinical effects. Urethral stricture did not recur in these patients for 7 to 8 months. There were no side effects in these patients. Sairei-to is suggested to be useful to prevent recurrence of urethral stricture after a transurethral procedure.

Transurethral incision (TUI) was performed as the initial treatment in 10 children with ureteroceles. Three patients had ureteroceles associated with a single ureter. TUI relieved hydronephrosis and preserved renal function in all 3 cases. Urinary tract infection developed in no patients. However, all the patients required an antireflux operation because of postoperative vesicoureteral reflux (VUR). Seven children had a total of 8 ureteroceles associated with a duplex system. TUI resulted in preservation of the upper pole function in 6 of the 8 ureteroceles. Urinary tract infections and VUR developed in 3 and 7 patients, respectively. Common sheath reimplantation was performed in 2 ureteroceles. TUI relieves obstruction before the onset of devastating infections although it carries the risk of postoperative VUR. We recommend TUI as the initial treatment for ureteroceles associated with both single and duplex systems.

bisacodyl tablets. The noninferiority margin was > 9.0%. If noninferiority criteria were satisfied, a test for superiority was performed. If the lower bound of the CI was >0%, superiority was demonstrated. Results: A total of 603 randomised patients self-administered either P/MC (n = 305) or PEG + 2 bisacodyl tablets (n = 298). P/MC was superior compared with PEG + 2 bisacodyl tablets in overall colon cleansing (CI: 3.4) and in cleansing of the ascending (CI: 4.9), mid (CI: 1.6), and recto-sigmoid (CI: 0.4) segments of the colon. Overall colon cleansing was successful in a greater number of patients receiving P/MC than PEG + 2 bisacodyl tablets (84.2% vs 74.4%). Similarly, the proportion of patients who were successfully cleansed was higher for P/MC than for PEG + 2 bisacodyl tablets in the ascending (89.5% vs 78.8%), mid (92.4% vs 85.9%), and recto-sigmoid (92.4% vs 87.2%) segments of the colon. The incidence of treatmentemergent adverse events (TEAEs) was similar between P/MC and PEG + 2 bisacodyl tablets (69.2% vs 72.8%). Commonly reported TEAEs attributed to P/MC or PEG + 2 bisacodyl tablets were nausea (2.6% vs 3.7%), vomiting (1.0% vs 3.4%), headache (1.6% vs 1.7%), and chills (0% vs 1.0%). Conclusions: P/MC administered as a split dose in preparation for colonoscopy is superior in colon cleansing and has a similar short-term safety profile compared with day-before dosing of PEG + 2 bisacodyl tablets.

We studied 15 resected cases with a history of apoplexy (2.5%) among 599 cases of esophageal cancer admitted between 1972 and 1993. Fourteen were male, and female, aged 48 to 77 years. Twelve had suffered from cerebral infarction, 2 intracerebral hemorrhage, and one subarachnoid hemorrhage. Duration from apoplexy to operation was between 2 months and 19 years in the cerebral infarction cases, between 8 and 10 years in the intracerebral hemorrhage cases and 4 years in the subarachnoid hemorrhage case. Preoperative neurological disturbance was found in 7 of the 12 cerebral infarction cases, and in both intracerebral hemorrhage cases. Four cases showed hemiplegia, and the other 5 cases showed partial paralysis of limbs. Preoperative complications were found in 7 of the 15 cases, and consisted of diabetes mellitus in 5, hypertension in 4, bronchial asthma in one, and renal dysfunction in one case. Intra- and postoperative complications were found in 11 of the 15 cases, and consisted of anastomotic leakage in 5, delirium in 3, apoplexy in 2, peritonitis in one, ARDS in one, intraoperative cardiac arrest in one, and wound infection in one. Postoperative disorders of consciousness were found in 5 cases, consisting of delirium in 3, and excitation at awakening of anethesia in 2 cases. Rate of direct operative death was 6.7% in preoperative apoplectic patients, and 8.5% in non-apoplectic patients, and there was no significant difference between the 2 groups. On the other hand, rate of postoperative apoplexy was 13.3% in the preoperative apoplectic patients, and 0.4% in non-apoplectic patients. There was a significant difference between them (p0.01). But they were cured of it, and left our hospital. It is concluded that active surgical treatment can be indicated for esophageal cancer patients with a history of apoplexy, if more attention is given to the management of diabetes mellitus or hypertension.

Transcription factor nuclear factor kappa B (NF kappa B) controls gene expression of a number of genes including cell adhesion molecules such as E-selectin, intercellular adhesion molecule 1, and vascular adhesion molecule 1. These cell adhesion molecules are known to play important roles in a critical step of tumor metastasis, arrest of tumor cells onto the venous or capillary bed of the target organ. NF kappa B is activated by extracellular signals such as those elicited by proinflammatory cytokines, tumor necrosis factor and interleukin 1 (IL-1). Here we demonstrate that IL-1 beta induces nuclear translocation of NF kappa B in human umbilical vein endothelial cells, followed by induction of cell surface expression of E-selectin, intercellular adhesion molecule-1, and vascular adhesion molecule 1, and subsequently augments adhesion of those cancer cells expressing sialyl Lewis X antigen, a ligand to E-selectin. We have also demonstrated that the adhesion of tumor cells to IL-1 beta-treated human umbilical vein endothelial cells can be inhibited by anti-NF kappa B reagents such as N-acetyl L-cysteine, aspirin, or pentoxifylline. These observations indicate the involvement of NF kappa B in cancer metastasis and the feasibility of using anti-NF kappa B reagents in preventing metastasis.

A case of an 81-year-old man with Fournier's gangrene was reported. The patient visited our hospital complaining of scrotal swelling and redness. Perirectal abscess was found and ultrasound study revealed thickness of scrotal skin and normal testes. The patient was immediately treated with antibiotics, incision and debridement of the scrotal skin. The lesion healed 8 weeks later.

A case of an inflammatory pseudotumor of the urinary bladder in a 46-yr-old man is presented. This rare, benign, and presumed non-neoplastic, reactive lesion must be differentiated from sarcomas of the urinary bladder. In the present case, we could demonstrate an inflammatory and reactive nature for the pseudotumor. Histologically, the presence of many Brunn's nests with infiltration of inflammatory cells and proliferation of capillaries in the myxoid stroma indicated a chronic inflammatory background for this lesion. It is apparent from morphology and immunohistochemistry findings that the proliferating spindle-shaped cell is of mesenchymal origin and not malignant in nature. These findings suggest that chronic inflammation can induce an overreaction of the bladder wall resembling tumor formation.