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Deals with loop-shaping characteristics of a human operator in a manual control system with a force reflective controlling device (controlling lever). The system consists of a dual loop. The inner loop is force based and the outer loop is visual based. In the inner loop, a hand motion control loop, the operator applies a force to the lever to bring its position close to a hand (or lever) position command. In the outer loop the operator generates the command from information of the visual sensation such as a target position. This paper focuses on the inner loop, that is the hand position control system, and investigates its structure and properties. As a result, loop-shaping characteristics of a human operator in the hand motion control system are discovered. The characteristics correspond to the crossover model, well-known operator's characteristics in manual control systems based on visual sensation. Further, it is confirmed that the discovered characteristics of the operator are explainable by the structure of the proposed model.

Develops basic techniques of an oceanographic vertical observation buoy system to keep the position within some area, such as a warm-core. The authors have tested experimentally a prototype of a dynamic positioning buoy with GPS. The authors describe in detail a self-returning program examination of the prototype.

In this paper, an autonomous distributed system consisting of multiple actuators loop-connected into a wheel shape was considered. In order to simplify the controller design procedure, genetic programming and genetic algorithms were employed to directly search for optimal torque functions of this multiactuator system of which nonlinearity and multiplicity of actuators make it difficult to determine the optimal torque functions analytically. As multiple torque functions corresponding to actuators in different states need to be obtained, multiple GP processes which share the same fitness value were executed in parallel. Though it is believed that GP is well suited to difficult control problems where no exact solution is known, at the population size of 50, commonly used GP failed to obtain an acceptable solution for this system. Step by step experiments demonstrate how GP with small population size can be used to improve a predetermined solution by using the proposed concepts of partially hinted initial population and common-branch crossover operator. Separately, another GA search method which manipulates directly on the values of the torque tables was employed and the simulation and experimental results of both methods were compared.

We investigated a resonant-type Mach-Zehnder optical modulator consisting of planar structures, designed to overcome the problems of making such a device and to increase the efficiency of the optical modulation. the electrode structure consists of modulating electrode and two stubs. The modulating electrode has two arms of the same length, and is an asymmetric coplanar waveguide.

To realize an efficient pre-production line, we developed "Autonomic Information System for modern LSIs " (AIS). This system is combined with yield management system (YMS) and manufacturing management system (Fab Host) by using personal computers. It distributes visual information about loss-time of bottleneck equipment on production clearly and a particle map with background data for each lot and each process. By using this information system, all the concerned persons (line manager, engineers and technicians) can dissolve problems independently, appropriately and quickly like the autonomic nervous system works in the human body. "AIS" has been developing in our R&D pilot line as a loss-time reduction tool.

An essential task for a multiple autonomous mobile robot system is to search for and to gather objects. To accomplish such a task effectively, it is essential that all the agents share common information on locations of objects, comrade agents and the place to deposit the objects. The authors have proposed a method called "COMPASS" to realize this only using two functions of each agent-to estimate the location of the objects by dead reckoning and to exchange the information with each other by one to one bidirectional proximity communication. In the paper, a new concept "figure of confidence (FOC)" is introduced to the COMPASS for higher performance and additional capability. The FOC is a self-estimated measure for accuracy of information in each agent. Using the FOC, agents can accelerate information sharing and handle multiple objects without significantly increasing complexity of the algorithm. The validity of FOC is confirmed through computer simulations.

Our experience suggests that there exist what may be termed 3 pedigrees of hereditary prostate cancer. Typical of pedigree 1, three brothers were affected by prostate cancer. A characteristic of this pedigree was the existence of some patients with bladder cancer and some with brain tumor. In pedigree 2, the paternal side of the family had been affected by prostate cancer for 3 generations. It was characteristic of this pedigree that the age at diagnosis was lower with each succeeding generation. In pedigree 3, a father and 2 sons were affected with prostate cancer. In 3 families, there were 9 patients with hereditary prostate cancer. The age at diagnosis ranged from 47 to 84 years, with an average age at diagnosis of 68.7 years, which was 5 years younger than the average age at diagnosis in Japan.

Resonant structures for band-operation optical modulators were investigated in this paper. Modulation efficiency was enhanced by the resonance of the modulating electrode. We can optimize the resonant structures by using equivalent circuit model and numerical electro-magnetic analysis such as the finite element method. The electric field distribution on the electrode was directly measured by the EO sampling technique.

We encountered a 60-year-old man with severe respiratory failure after CABG. The patient received veno-venous extracorporeal membrane oxygenation (ECMO) in which blood was drained from the right atrium through a cannula and was returned via a cannula in the left femoral vein. During ECMO, the flow rate was 2.8-3.0 l/min (about 40 ml/kg/min) and the assist was needed for 5 years. Veno-venous ECMO is a simple and effective therapy for acute respiratory failure unresponsive to conventional treatment.

Although the projectile fragment separator (RIPS) at the RIKEN accelerator facility (RARF) provides wide variety of energetic radioactive beams, it is not adequate for low energy beam experiments such as precision spectroscopy of trapped ions. Aiming for an efficient deceleration and cooling of energetic radioactive ions, the development of an rf ion guide system which comprises a large He gas cell and an rf funnel structure in the cell is under progress. A proof of the principle (pop) machine has been successfully tested on-line for collection of 70-MeV/u 8Li ions. A compact gas cell of 70 cm in length and 10 cm in diameter with a He gas pressure of 30 Torr was placed after a variable wedge shaped energy degrader. The ions were stopped in the gas and were transported by a superposition of dc and rf electric field toward a small exit hole. The overall efficiency was 10−4 which can be separated into a gas stopping efficiency of 0.43% and an ion-guide efficiency of 2.4%. The former is limited by the volume ...

Purpose: To assess the effect of audio instruction on tracking accuracy for MHI‐TM2000 (VERO).Methods: Anterior‐posterior abdominal skin surface displacements for ten patients with a lungtumor were measured by a respiratory monitoring system with and without audio instruction. Based on the obtained twenty respiratory patterns, a three‐dimensional movable phantom with a steel ball target of 9.5 mm in diameter was driven along the superior‐inferior direction. The target position was measured in real time by the laser displacement gauge and the orthogonal kV x‐ray imaging subsystem. Simultaneously, the kV x‐ray imaging subsystem predicted the future target position, and then the system controller calculated the corresponding rotation angles. Thereafter, the gimbaled x‐ray head tracked the target based on the command. Total tracking system error (ET) was computed by both target prediction errors and mechanical response errors of the gimbaled x‐ray head, and then compared between with and without audio instruction.Results: By applying audio instruction, the reproducibility of respiratory pattern was significantly improved (p < 0.01), although motion amplitude was increased by a factor of 2.1 on average. The root mean squares of the ET (RMSET) were up to 1.18 mm without audio instruction and 1.26 mm with audio instruction, respectively. There was no significant difference between free breathing and audio instruction in the RMSET (p = 0.26). The correlation between the RMSET and the 90th percentile of respiratory speed (v) was stronger under the conditions of audio instruction (R2 = 0.77) than under the conditions of free breathing (R2 = 0.29) Conclusions: Audio instruction improved reproducibility of respiratory pattern but increased to the v that negatively affected the tracking accuracy. The tracking errors can be deduced from the v for regular respiratory pattern, suggesting upper limit of the v to achieve high tracking accuracy. This research was in part sponsored by Mitsubishi Heavy Industries, Ltd., Japan.

Incidence of Radiation Pneumonitis After Multiple Courses of Stereotactic Body Radiation Therapy for Lung Cancer Patients T. Yoshitake, K. Nakamura, T. Sasaki, S. Ohga, K. Terashima, K. Asai, K. Matsumoto, H. Hirata, M. Inamori, Y. Matsuo, T. Kamitani, Y. Shioyama, M. Shinoto, H. Hirata, and H. Honda; Department of Comprehensive Clinical Oncology, Graduate School of Medical Sciences, Kyushu University, Fukuoka City, Japan, Department of Clinical Radiology, Graduate School of Medical Sciences, Kyushu University, Fukuoka, Japan, Kyushu International Heavy Ion Beam Therapy Center, Tosu, Japan, Department of Health Sciences, Graduate School of Medical Sciences, Kyushu University, Fukuoka, Japan

Antagonist activities of (R)-(-)-2-(benzo[1,3]dioxol-5-yl)-N-(4-isopropylphenylsulfonyl)-2-(6-methyl- 2-propylpyridin-3-yloxy) acetamide hydrochloride (CAS 188710-94-3, PABSA), a novel endothelin (ET) receptor antagonist, for ETA and ETB receptors were evaluated using rat aortic smooth muscle A7r5 cells and isolated rat thoracic aorta. PABSA concentration-dependently inhibited the ET-1-induced increase in intracellular calcium concentration ([Ca2+]i) mediated via ETA receptors in A7r5 cells with an IC50 of 0.17 nmol/l. PABSA antagonized the ETA receptor-mediated contraction induced by ET-1 in endothelium-denuded rat aorta with a Kb of 0.74 nmol/l. The potency of PABSA in inhibiting ETA receptor-mediated vasocontraction was approximately 40- and 100-fold greater than those of BQ-123, a selective ETA antagonist, and bosentan, a mixed ETA/ETB receptor antagonist, respectively. ETB receptor-mediated endothelium-dependent vasorelaxation induced by ET-3 in the aorta was also antagonized by PABSA, with a Kb of 9.8 nmol/l. In contrast, PABSA affected neither the vasocontraction induced by KCl or norepinephrine nor the vasorelaxation induced by acetylcholine or prostaglandin I2 in the aorta. These results suggest that PABSA is a highly potent and selective ETA receptor antagonist.

To evaluate the mechanism of renal transport of quinolone antibacterial drugs, we examined the interaction of levofloxacin with p-aminohippurate (PAH) transport systems and the transport of levofloxacin in renal epithelial cells.Transport of [14C]PAH or [14C]levofloxacin was measured using OK cell monolayers grown on microporous membrane filters.Transcellular transport from the basolateral to the apical side and cellular accumulation of [14C]PAH were inhibited by levofloxacin. Both the initial uptake of [14C]PAH from the basolateral side and the efflux to the apical side were inhibited by levofloxacin. The basolateral-to-apical transcellular transport of [14C]levofloxacin was greater than that in the opposite direction. [14C]Levofloxacin efflux to the apical side was greater than that to the basolateral side. Unlabeled levofloxacin and grepafloxacin inhibited the transcellular transport of [14C]levofloxacin, accompanied by an increase of cellular accumulation. However, neither PAH nor an anion transport inhibitor 4-4'-diisothiocyanostilbene-2,2'-disulfonic acid (DIDS) affected the basolateral-to-apical transport of [14C]levofloxacin nor its uptake from the basolateral side.These results indicated that levofloxacin inhibits PAH transport across both the basolateral and apical membranes of OK cells, but are not transported via the systems for PAH transport. The existence of a specific transport system for quinolones was indicated in OK cells.

Although human lung adenocarcinoma has diverse histological subtypes, the correlation between histological subtypes and occurrence of the p53 gene mutation has been given less attention. We investigated 145 surgically resected lung adenocarcinomas to search for the incidence of p53 mutations and for record data on survival in each histological subtype, according to the new WHO criteria (1999). The frequency of p53 mutation in bronchioloalveolar carcinoma (BAC; 0% in 17 cases) and BAC with invasive growth component (BAC-invasive; 11% in 27 cases), which is conventionally categorized as the mixed subtype in WHO typing, were apparently significantly lower than in other types (non-BAC including acinar, papillary, solid, or mixed histology with these subtypes; 48% in 101 cases; P0.01). Multivariate analysis revealed that the histological subtype including BAC-invasive was a strong, independent, and significant prognostic factor (P0.03), as were tumor size and pathological stage (P0.001 and 0.002, respectively) for overall survival. However, the occurrence of p53 mutation itself was seen to be significant only in case of the univariate analysis. Therefore, histological subtyping may be a better prognostic indicator than is p53 mutation. These findings suggest that the WHO classification with the BAC and BAC-invasive from other histological subtypes may prove useful to predict the outcome for surgically treated patients with lung adenocarcinoma.

Here we report a case of spinal dural arteriovenous fistula(AVF) treated by endovascular embolization. A 58-year-old female presented with progressive intermittent claudication and numbness of the lower extremities. MRI showed swelling of the spinal cord with intramedullary high signal intensity on T2-weighted image and intramedullary enhancement, suggested spinal cord myelopathy. Myelography demonstrated the dilated serpentine vessels in the subarachnoid space and focal filling defect. Angiography showed spinal dural AVF fed by bilateral lateral sacral artery. The draining vein was posterior spinal vein. Endovascular embolization using liquid material was performed under general anesthesia. The injection of glue included the distal feeding artery, the shunt itself and the initial part of draining vein. A complete cure was achieved, with a normal postoperative angiogram. MRI returned to normal with complete disappearance of T2 high signal, cord enlargement and enhancement by contrast medium. It was suggested that venous congestion induced the transient spinal ischemia, manifested as intermittent claudication. Endovascular embolization using liquid material was safe and quite effective for spinal dural AVF.

The RIKEN Mouse Gene Encyclopaedia Project, a systematic approach to determining the full coding potential of the mouse genome, involves collection and sequencing of full-length complementary DNAs and physical mapping of the corresponding genes to the mouse genome. We organized an international functional annotation meeting (FANTOM) to annotate the first 21,076 cDNAs to be analysed in this project. Here we describe the first RIKEN clone collection, which is one of the largest described for any organism. Analysis of these cDNAs extends known gene families and identifies new ones.

Human bone marrow stroma (BST)-dependent myeloma sister cell lines MOLP-6 and MOLP-7 were established from the peripheral blood of a multiple myeloma (MM) patient with IgA kappa type MM (stage IIIB). The growth of the cell lines is constitutively dependent on BST cells; none of the cytokines tested nor the culture supernatant of the BST cells could support the growth. Both cell lines showed typical plasma cell morphology with abundant cytoplasm and one to four nuclei under Wright staining. The immunoprofiles of MOLP-6 and MOLP-7 correspond to that seen typically in primary MM cells: positive for cytoplasmic immunoglobulin (Ig) chains, a heavy and kappa light chains, CD9, CD28, CD40, CD44, CD45, CD56, and PCA-1; the cells were negative for surface Igs and various other B-cell, T-cell and myelomonocyte associated markers. Both cell lines also expressed adhesion molecules including HCAM (CD44), VLA-4 (CD49d/CD29), VLA-6 (CD49f/CD29), ICAM-1 (CD54), NCAM (CD56), LFA-3 (CD58) and L-selectin (CD62L). The doubling time of MOLP-6 and MOLP-7 was 48 and 168 hours, respectively. In addition to this growth characteristic, the maximum cell density of each cell line was obtained at 1.7 x 10(6) cells/ml and 9.7 x 10(5) cells/ml, respectively. The characteristics of each cell line may reflect intraclonal variation of the proliferative capacity. The MOLP-6 together with the MOLP-7 sister will be useful model systems for the investigation of the biology of myeloma.

The development and progression of diabetic nephropathy is dependent on glucose homeostasis and many other contributing factors. In the present study, we examined the effect of nitecapone, an inhibitor of the dopamine-metabolizing enzyme catechol-O-methyl transferase (COMT) and a potent antioxidant, on functional and cellular determinants of renal function in rats with streptozotocin-induced diabetes. Administration of nitecapone to diabetic rats normalized urinary sodium excretion in a manner consistent with the dopamine-dependent inhibition of proximal tubule Na,K-ATPase activity. Hyperfiltration, focal glomerulosclerosis, and albuminuria were also reversed by nitecapone, but in a manner that is more readily attributed to the antioxidant potential of the agent. A pattern of elevated oxidative stress, measured as CuZn superoxide dismutase gene expression and thiobarbituric acid-reactive substance content, was noted in diabetic rats, and both parameters were normalized by nitecapone treatment. In diabetic rats, activation of glomerular protein kinase C (PKC) was confirmed by isoform-specific translocation and Ser23 phosphorylation of the PKC substrate Na,K-ATPase. PKC-dependent changes in Na,K-ATPase phosphorylation were associated with decreased glomerular Na,K-ATPase activity. Nitecapone-treated diabetic rats were protected from these intracellular modifications. The combined results suggest that the COMT-inhibitory and antioxidant properties of nitecapone provide a protective therapy against the development of diabetic nephropathy.

Carp homologues of p38 mitogen-activated protein kinase (MAPK) and its activator MAPK kinase 6 (MAPKK6, referred to as MKK6) were identified. There exist at least two distinct carp p38s, cp38a and cp38b, both of which consist of 361 amino acids. The transcript of c38a was exclusively expressed in the ovary, whereas that of cp38b was ubiquitously expressed. Western blot analysis with anti-(phosphorylated MAPK) Ig specific to the active p38 or JNK has shown that p38 was activated in response to hypertonic stress (1 M sorbitol) in epithelioma papilosum cyprini carp epithelial cells (EPC) and that the activation of p38 proceeded faster to the maximal level than that of JNK. Carp homologue (cMKK6) of p38 activator MKK6 consists of 404 amino acids. It was expressed ubiquitously but was most abundant in the ovary. An in vitro kinase assay demonstrated that cMKK6 is an upstream activator of cp38 and cp38b in carp because it specifically phosphorylated and activated cp38a and cp38b. Interestingly, we found that cMKK6 has a nuclear export signal (NES) sequence in its N-terminal region although upstream activators of stress-activated MAPKs, p38 and JNK, do not in other animals. The NES sequence facilitated nuclear export of cMKK6 and ovalbumin. Leucine residues in the sequence were crucial for the NES activity, as the activity was lost on replacement of the leucines to alanines. The existence of an NES in cMKK6 implies the requisite of strict regulation of the p38 MAPK pathway in carp. The abundance of these components for the stress-activated pathway in the ovary might be related to ectogenetic early development.

The novel multiple myeloma (MM) cell line MOLP-5 and its homologous sister cell line B407, a lymphoblastoid cell line (LCL), were established from the peripheral blood of a 71-year-old Japanese patient with Bence-Jones kappa-type multiple myeloma (stage IIIB with hyperammonaemia and hypercalcaemia). The growth of MOLP-5 cells is constitutively dependent on bone marrow stroma (BST) cells; none of the cytokines tested nor the culture supernatant of the bone marrow stroma cells could support the growth of MOLP-5. Wright-Giemsa-stained MOLP-5 cells showed typical plasma cell morphology with abundant cytoplasm and one to three nuclei. The immunoprofile of MOLP-5 corresponds to that seen typically in primary MM cells: positive for cytoplasmic immunoglobulin (Ig) kappa light chain, CD28, CD29, CD38, CD40, CD44, CD49d, CD54, CD56, CD58, CD71, CD138 and PCA-1; the cells were negative for surface Ig and various other B-cell, T-cell and myelomonocyte-associated immunomarkers. Interleukin 6 (IL-6) receptor mRNA was found in the reverse transcriptase polymerase chain reaction (RT-PCR) analysis. IL-6 and IL-10 could induce cellular proliferation in short-term induction experiments. IL-6 or IL-10 production was not detected by specific enzyme-linked immunoabsorbent assay (ELISA). MOLP-5 cells expressed parathyroid hormone-related protein (PTHrP) at the mRNA level. Cytogenetic analysis showed the typical t(11; 14) chromosome abnormality. The novel MOLP-5 cell line together with the B407 B-LCL sister line will be useful model systems in the investigation of the biology of MM.

Some plasma membrane receptors in yeast are known to be internalized and degraded in lysosomes upon ligand-dependent ubiquitination. However, the role of ubiquitination in endocytosis and lysosomal degradation in higher eukaryotes has been controversial. In order to directly assess this question, we investigated the fate of chimeric molecules in which ubiquitin moiety was fused in-frame to the cytoplasmic region of membrane proteins. The chimeric proteins with the wild-type ubiquitin were endocytosed and delivered to lysosomes efficiently. Mutant ubiquitin with lysine-to-arginine substitution could still mediate endocytosis, suggesting that polyubiquitination is not required for the endocytosis. We next searched for the existence of an endocytosis signal(s) in the ubiquitin moiety, and identified a di-leucine signal, Leu(43)-Ile(44). The Leu(43)-Ile(44) sequence mediated endocytosis and lysosomal sorting in a Leu(43)-dependent manner. These results suggest that the di-leucine signal in ubiquitin can be involved in ubiquitination-mediated endocytosis and lysosomal targeting of membrane proteins.

Malignancies involving natural killer (NK) cells are rare disorders. The complexity of NK cell-involving disorders has only recently been appreciated. Modern classifications discern immature (precursor) from mature NK cell leukemias-lymphomas. Continuous NK leukemia-lymphoma cell lines represent important model systems to study these neoplasms. While there are a number of putative NK cell lines which are, however, either not characterized, not immortalized, non-malignant, non-NK, or plain false cell lines, six bona fide malignant NK cell lines have been established and are sufficiently well characterized: HANK1, KHYG-1, NK-92, NKL, NK-YS and YT. Except for YT which was derived from a not further defined acute lymphoblastic lymphoma, these cell lines were established from patients with various NK cell malignancies. Five of the six cell lines are constitutively interleukin-2-dependent. Their immunoprofile is remarkably similar: CD1-, CD2+, surface CD3 (but cytoplasmic CD3epsilon+), CD4-, CD5-, CD7+, CD8-, CD16-, CD56+, CD57-, TCRalphabeta-, TCRgammadelta-, negative for B cell and myelomonocytic markers. The immunoglobulin heavy chain and T cell receptor genes are all in germline configuration. All six lines show complex chromosomal alterations, with both numerical and structural aberrations, attesting to their malignant and monoclonal nature. Functionally, these cells which contain azurophilic granules in their cytoplasm are nearly universally positive in NK activity assays. Three of five cell lines are Epstein-Barr virus-positive (type II latency). The composite data on these six cell lines allow for the operational definition of a typical malignant NK cell line profile. NK leukemia-lymphoma cell lines will prove invaluable for studies of normal and malignant NK cell biology.

The angiographic assessment of coronary stenosis has many limitations, especially after coronary intervention. To determine the physiologic significance of such lesions, we measured the mean translesional pressure gradients at rest (P1), those during hyperemia induced by intracoronary administration of papaverine (12 mg in the left and 8 mg in the right coronary artery) (P2), and fractional myocardial flow reserve (FFRmyo) which is derived from the ratio of the mean distal coronary pressure and aortic pressure during hyperemia. Our objective was to determine the relations among P1, P2, and FFRmyo and restenosis as potential predictors of chronic restenosis. The grouped study consisted of 32 patients with ischemic heart disease scheduled to undergo intervention. The distal coronary pressure was measured using a 0.014 inch pressure monitoring wire (Pressure Guide, Radi Medical Systems, Uppsala, Sweden). The guidewire was advanced through the lesional segment. The mean percent diameter stenosis (%DS) was 67.9 +/- 13.3% before intervention and 22.9 +/- 15.3% after intervention. The mean P2 (9.9 +/- 5.3 mmHg) was significantly higher than the mean P1 (4.5 +/- 3.6 mmHg; p0.0001). There was no correlation of P1, P2 or FFRmyo with the %DS after intervention. Follow-up angiography (after 6 months) performed on all patients revealed restenosis (%DSor =50%) in four patients (12.5%). There was no correlation between P1 and the %DS at the follow-up angiography, but the relation between P2, FFRmyo and the %DS at the follow-up angiography was significant (r = 0.599, p0.01; r = 0.703, p0.0001, respectively). As a measurement of P2, FFRmyo is useful for the determination of the endpoint of intervention in consideration of prevention of restenosis. A new endpoint may be established after further evaluation in a greater number of patients.

In order to identify cross-culture contamination of cell lines, we applied DNA fingerprinting using variable number of tandem repeat (VNTR) loci and short tandem repeat (STR) loci amplified by polymerase chain reaction (PCR) instead of a radioisotope labeled multilocus probe. Eleven cell lines were used for the Apo B and D1S80 loci detection, and twelve cell lines were examined in the Y-chromosome analysis. The data obtained from the sister cell lines NALM-6 and B85, two MOLM-1 cultures from two cryopreserved tubes, and four subclones of BALM-9 and its sister cell line BALM-10, displayed clear and distinct bands of each PCR product for both Apo B and D1S80. Detection of a Y-chromosome DNA sequence is another very informative marker for the identification of cell lines, if the Y-chromosome is present. We examined eight cell lines for the expression of four STR loci; the data thus generated were compared with the results previously reported from other laboratories. The resulting electrophoretic banding patterns showed that our "home-made" STR detection system is a useful and efficient tool for the authentication of cell lines. PCR detection of VNTR and STR loci represents a simple, rapid and powerful DNA fingerprinting technique to authenticate human cell lines and to detect cross-culture contamination. This PCR technique may be used in lieu of the more time-consuming, labor-intensive and radioactive Southern blot multilocus method.

Alteration of the p53 gene is one of the most frequent events in human tumorigenesis. The inactivation of p53 tumor suppressor function can be caused by chromosome deletion, gene deletion, or mainly by point mutations. p53 mutations occur moderately often in hematopoietic malignancies. A significantly higher frequency of p53 alterations in cell lines vs primary samples has been observed for all types of malignant hematopoietic cell lines. It has been postulated that p53 gene abnormalities arise in cell lines during in vitro establishment of the culture or prolonged culture; but it is also conceivable that those cases that carry p53 mutations may be more suitable for in vitro establishment as permanent cell lines. We analyzed data on the p53 gene status in a panel of matched primary hematopoietic tumor cells and the respective cell lines derived from this original material. In 85% (53/62) of the pairs of matched primary cells and cell lines, the in vivo and in vitro data were identical (both with p53 wild-type or both with the same p53 mutation). In some instances, serial clinical samples (eg at presentation and relapse) and serial sister cell lines were available. These cases showed that a clinical sample at presentation often had a p53 wild-type configuration whereas the derived cell line and a relapse specimen carried an identical p53 point mutation. These findings suggest that a minor clone, at first undetectable by standard analysis, represents a reservoir for the outgrowth of resistant cells in vivo and also a pool of cells with a growth advantage in vitro, providing a significantly higher chance of immortalization in culture. This was further supported by studies employing mutant allele-specific gene amplifications, a technique which is significantly more sensitive (100- to 1000-fold) than the commonly applied SSCP assay with a sensitivity threshold of about 10% mutated cells within a pool of wild-type cells. Taken together, this analysis confirms the usefulness of human hematopoietic cell lines as in vitro model systems for the study of the biology of hematopoietic malignancies. It further underlines the notion that p53 gene alterations confer a survival advantage to, at least some, malignant cells in vitro and presumably also in vivo; however, it is highly unlikely that a p53 mutation alone would suffice for the immortalization of a cell line in vitro or tumor development in vivo. Leukemia (2000) 14, 198-206.

The relationship between the net charge of molecules and their mobility on electrophoresis was analyzed for Drosophila alpha-amylases. Most of the differences in electrophoretic mobility, 98.2%, can be explained by the charge state. Therefore five reference amino acid sites, which are informative residues for charge differences among amylase isozymes, were considered for the evolution of the isozymes in Drosophila melanogaster. The amylase isozymes in D. melanogaster can be classified into three groups, I (AMY1, AMY2, and AMY3-A), II (AMY3-B and AMY4), and III (AMY5, AMY6-A, and AMY6-B), based on the differences in the reference sites. The most primitive amylase in D. melanogaster was found to belong to Group I, most likely the AMY2 isozyme. Groups II and III could have been derived from Group I. These results were confirmed by the analysis of 38 amino acid sites with charge differences in Drosophila.

We discuss tachyon configuration for the unoriented bosonic string theory which produces a bosonic string theory with SO(32) gauge symmetry in ten dimensions. It is closely related to the tachyon condensation scenario proposed by A. Sen. We also give the boundary state description of the tachyon condensation process, with some emphasis on the r\^ole of orbifold conformal field theory., Comment: Proceeding of the talk given at YITP workshop ``Tachyon condensation and boundary state''(October 27-29, 1999), latex file

Microwave spectroscopy in highly excited Rydberg states is proposed for structure determination of small metal cluster ions. An experimental setup for microwave optical triple resonance is developed and tested for Yb atom. Many microwave transitions are measured with the line width less than 1 MHz (HWHM).

For the purpose of achieving gene transfer into cells mediated by peptides with a short chain length, we employed two kinds of amphiphilic alpha-helix peptides, mastoparan (INLK-ALAA-LAKK-IL-NH2) obtained from wasp venom and an alpha-helix model peptide (LARL-LARL-LARL-NH2). Furthermore, to strengthen the hydrophobicity of the peptide required for the formation of the aggregates with the DNA, we modified these peptides using several lipophilic groups, i.e. acyl groups with a single chain, a dialkylcarbamoyl group and a cholesteryloxycarbonyl group. We examined the ability of the peptides and their derivatives to bind and aggregate with plasmid DNA, the structural change in the peptides caused by binding with the DNA and the in vitro gene transfer abilities into COS-7 cells. As a result, mastoparan was found to acquire the DNA binding ability by introduction of the lipophilic group. The conformational change in the peptides depended on the hydrophobicity of the introduced acyl group. The DNA complex of most lipophilic mastoparan derivatives could be incorporated into the cells via the endocytosis pathway. In the case of the helix model peptide, the acyl group with a moderate chain length was required for the formation of the aggregate which is competent for incorporation into the cells. In this study, we succeeded in giving such short peptides sufficient gene transfer ability by modifying them with some lipophilic groups. However, the influence of the modification by the lipophilic groups on the formation of aggregates with DNA and the gene transfer ability depended on the structure of the peptide portion. These results indicate that consideration of total hydrophobicity balance is needed for the design of an efficient gene carrier peptide.

We present a panoptic survey of cell line cross-contamination (CLCC) among original stocks of human cell lines, investigated using molecular genetic methods. The survey comprised 252 consecutive human cell lines, almost exclusively tumor-derived, submitted by their originators to the DSMZ and 5 additional cell repositories (CRs), using a combination of DNA profiling (4-locus minisatellite and multilocus microsatellite probes) and molecular cytogenetics, exploiting an interactive database (http://www.dsmz.de/). Widespread high levels of cross-contaminants (CCs) were uncovered, affecting 45 cell lines (18%) supplied by 27 of 93 originators (29%). Unlike previous reports, most CCs (42/45) occurred intraspecies, a discrepancy attributable to improved detection of the more insidious intraspecies CCs afforded by molecular methods. The most prolific CCs were classic tumor cell lines, the numbers of CCs they caused being as follows: HeLa (n = 11), T-24 (n = 4), SK-HEP-1 (n = 4), U-937 (n = 4) and HT-29 (n = 3). All 5 supposed instances of spontaneous immortalization of normal cells were spurious, due to CLCC, including ECV304, the most cited human endothelial cell line. Although high, our figure for CCs at the source sets a lower limit only as (i) many older tumor cell lines were unavailable for comparison and (ii) circulating cell lines are often obtained indirectly, rather than via originators or CRs. The misidentified cell lines reported here have already been unwittingly used in several hundreds of potentially misleading reports, including use as inappropriate tumor models and subclones masquerading as independent replicates. We believe these findings indicate a grave and chronic problem demanding radical measures, to include extra controls over cell line authentication, provenance and availability.

Sclerosing stromal tumor is a rare ovarian neoplasm. We describe the radiologic findings of sclerosing stromal tumor in two patients. In both patients, MR and CT images showed a large mass in the left adnexal region. On dynamic contrast-enhanced images, the tumors showed early peripheral enhancement with centripetal progression.

The regulation of amylase activity in three Drosophila species, D. melanogaster, D. funebris and D. saltans, was analyzed by measuring the specific activity levels in four dietary environments, cornmeal, glucose, 5% starch, and 10% starch, at three developmental stages, i.e., the third-instar larval, pupal, and 2-day-old adult stages. The developmental profiles of amylase activity for the three Drosophila species showed that the level of activity was high at the larval and adult stages but substantially low at the pupal stage, suggesting that Drosophila does not utilize starch at the pupal stage. Divergence in the regulation of amylase was observed among the three Drosophila species on the following points. (1) The order of amylase specific activity was D. melanogasterD. funebrisD. saltans. (2) The response pattern to the dietary environment varied among the species and changed during development. (3) The timing of the switch in the response pattern to the dietary environment during development was before pupation in D. funebris and D. saltans but after pupation in D. melanogaster. The significance of the divergence in the regulation of amylase activity for adaptation to a starch environment in Drosophila is discussed.

The adaptation to glucose and starch foods in six species, D. melanogaster, D. virilis, D. saltans, D. funebris, D. levanonensis and D. americana, was studied by measuring productivity. D. melanogaster and D. virilis adapted more to the starch environment than to the glucose environment, while D. saltans adapted more to the glucose environment than to the starch environment. D. funebris, D. levanonensis, and D. americana did not distinctly adapt to either environment. In addition, the regulation of amylase in the six species was investigated by measuring the levels of amylase activity with glucose and starch food environments. The levels of amylase activity in D. levanonensis and D. saltans were substantially low, indicating that these species cannot utilize starch as a carbon source. The starch-adapted species, D. melanogaster and D. virilis, showed higher levels of amylase activity with the starch environment and higher inducibility. These results suggest that changing the regulation of amylase is important for the adaptation to a starch environment in Drosophila.

The p73 gene, a member of the p53 family, is a new candidate tumor suppressor gene. To investigate the possibility of genetic alteration of p73 in leukemia and lymphoma, we examined 55 cell lines and 39 patient samples together with 17 nonhematopoietic cancer cell lines. Gene expression of p73 was detected by reverse transcriptase-polymerase chain reaction (RT-PCR) in cell lines (5 of 7 pre B/B-acute lymphoblastic leukemia [ALL], 13 of 21 T-ALL/lymphoblastic lymphomas [LBL], 9 of 10 B-non-Hodgkin's lymphomas [B-NHL], 8 of 9 acute myelogenous leukemias [AML], 2 of 2 T-NHL, 3 of 3 multiple myeloma), and in patient samples (16 of 23 pre B-ALL, 5 of 8 T-ALL/LBL, 5 of 8 B-NHL). PCR-single-strand conformation polymorphism (SSCP) of cDNAs showed no mutation in 43 p73-expressing cell lines within the regions that corresponded to the 5 mutational hotspots of the p53 gene. Neither homologous deletion nor rearrangement of the p73 gene were found by Southern blot analysis in any of the cell lines that lack expression of p73. In contrast to prior published data, analysis of a polymorphic site showed that the p73 gene was expressed biallelically in cell lines and normal peripheral blood. Notably, the p73-negative cell lines were hypermethylated at a CpG island in the 5' untranslated region of the p73 mRNA, and treatment of these cell lines with 5-azacytidine (5-AC), a demethylation reagent, induced p73 expression. Taken together, we found that a sizable proportion (32%) of ALL/B-NHL cell lines and primary tumors had negligible or limited expression of the p73 gene associated with hypermethylation of the gene. These findings suggest that silencing of the p73 gene by hypermethylation may contribute to development and/or progression of lymphoid neoplasms.

Following preoperative administration of 5'-deoxy-5-fluorouridine (5'-DFUR) in patients with colorectal cancer, histologic antitumor effects as well as pyrimidine nucleoside phosphorylase (PyNPase) activities in cancerous tissues were examined. Fifty-five patients were randomly divided into two groups: Twenty-five patients received 5'-DFUR orally at a daily dose of 800 mg for 3 weeks before operation (Group A) and twenty-nine received no medication (Group B). PyNPase activities in cancerous tissues obtained from resected specimens in Group A and B were 49.7 +/- 21.6 and 66.6 +/- 40.0 U/mg prot, respectively, which were significantly higher than 25.5 +/- 14.1 and 22.1 +/- 11.2 U/mg prot in normal colorectal tissues. No significant difference was observed in PyNPase activities in cancerous tissues between Group A and B. PyNPase activities in cancerous tissues in Group A were no different either from 59.2 +/- 40.0 U/mg prot in biopsy specimens before administration of 5'-DFUR. Histologic anti-tumor effects were as follows: Grade 2, 1 case Grade 1b, 5; Grade 1a, 10; and Grade 0, 9. Although no correlation was observed between antitumor effects and PyNPase activities in cancerous tissues, antitumor effects of 2 cases in which PyNPase activities in cancerous tissues before administration of 5'-DFUR showed more than 100 U/mg prot were Grade 2 and 1b, respectively.

Chromosomal aberrations and inactivation of tumour suppressor genes are frequent in acute leukaemia. To determine whether the FHIT gene is involved in the development of leukaemia, we examined the FHIT transcript in 65 leukaemia cell lines, 5 fresh acute leukaemia patients at diagnosis and in complete remission, normal peripheral blood lymphocytes obtained from 14 healthy volunteers and Epstein-Barr (EB) virus transformed 5 B-cell lines (EB-lines), using nested reverse transcription-polymerase chain reaction and direct sequencing. The transcripts were classified into 4 patterns: pattern I revealed the normal transcripts only, pattern II the altered transcripts in addition to the normal transcripts, pattern III the altered transcripts without the normal transcripts and pattern IV an absence of normal and altered FHIT transcripts. Nineteen cell lines were classified as pattern I, 32 as pattern II, 2 as pattern III and 12 as pattern IV. The frequency of loss of FHIT expression (pattern III or IV) varied in each type of leukaemia cell line; the order ranked from the highest incidence was acute myeloid leukaemia (AML), T-cell acute lymphoblastic leukaemia (T-ALL), B-precursor ALL, B-ALL, and chronic myeloid leukaemia (CML). No genomic rearrangement was found in any samples examined. All of 5 patients showed same pattern II FHIT transcripts at 2 different stages of the disease. All normal peripheral blood lymphocytes and EB-lines were classified as pattern I or II. Our results suggested that patterns III and IV of FHIT transcripts might be associated with the development of a subset of leukaemia, while pattern II which has so far been reported as an aberrant transcript in varieties of malignant tumours might not be associated with leukaemogenesis.

We established three sister cell lines, NALM-30, NALM-31 and NALM-32, with biphenotypic features carrying myeloperoxidase mRNA and protein with complex Philadelphia (Ph) chromosome, t(9;22;10)(q34;q11;q22), from a patient with Ph-positive acute leukemia in relapse. Epstein-Barr virus nuclear antigen was negative. The morphological appearance of the cell lines is that of immature lymphoid cells. Expression of myeloid- and lymphoid-associated surface membrane antigens on these cells was detected allowing for the classification of "biphenotypic" leukemia. Immunophenotypically, the established cell lines reported here fulfill the European Group for the Immunological Characterization of Leukemias (EGIL) criteria for B-lineage derivation, however, surface and cytoplasmic immunoglobulin chains were negative. Whereas TGF-beta R (CD105), MCSFR (CD115), SCFR (CD117), IL-4R/IL-13R (CD124) and IL-6R (CD126) were not expressed, the cell lines were mostly positive for IFN-gamma R (CD119), IL-7R (CD127) and FLT-3R (CD135). The NALM-30, NALM-31 and NALM-32 cell lines together with their serial sister cell lines NALM-27 and NALM-28 which were established from the same patient at diagnosis provide unprecedented opportunities for studying a multitude of biological aspects related to normal and neoplastic immature B-lymphocytes.

Continuous human malignant hematopoietic (MH) cell lines have become invaluable tools for hematological diagnosis and research. Over the last 35 years several hundred cell lines spanning almost the whole spectrum of hematopoietic cell lineages have been described. The cardinal features of MH cell lines are their monoclonal origin, arrest of differentiation, genetic alterations, and unlimited proliferation; the major advantages of cell lines are the unlimited supply of cell material and the infinite storability and recoverability at will of the cells. Categorization of cell lines usually follows the physiological stages of hematopoietic differentiation in the various cell lineages. For an adequate classification, a detailed and comparative characterization of both primary and cultured cells is absolutely necessary. New cell lines, in particular, must be adequately characterized. While clinical and cell culture data and immunological and cytogenetic features are the most important data, cell lines should be described in as much additional detail as possible allowing any singular features to be pointed out. In addition to detailed characterization, immortality of the culture, proof of neoplasticity, authentication of the true origin of the cells, scientific significance and availability of the cell line for other investigators are of paramount importance. In summary, MH cell lines have the potential to greatly facilitate diverse studies of normal and malignant hematopoiesis; to that end, these cell lines must be extensively characterized and adequately described.

We reviewed the medical records and images of 39 women with palpable breast masses, including 18 cases with breast cancer and 21 cases with benign breast mass, to compare diagnostic accuracy and clinical agreement between ultrasonography and mammography in breast cancer detection. We showed that the operating characteristics of ultrasonography, including sensitivity, specificity and positive likelihood ratio, were superior to that of mammography. We also confirmed that the established malignant criteria for malignancy, including lobulated shape, irregular contour and non-uniform internal echo in ultrasonography, and stellate mass in mammography, were statistically significant in distinguishing breast cancer from benign breast lesion. Moreover, the interobserver agreement kappa of ultrasonography and mammography were 0.64 and 0.59, respectively. Ultrasonography was also superior to mammography in attaining clinical agreement between the different observers in breast cancer detection.

The benefits of early extubation and assisted ventilation with mini-tracheostomy and preventive positive pressure ventilation in the early stage after thoracic esophageal cancer operation are examined and compared. Subjects were 23 patients who underwent radical operation for thoracic esophageal cancer in our hospital over a 5 year period. Ten patients (A group) underwent positive pressure ventilation with postoperative intubation for a certain period. Thirteen patients (B group) were extubated the day after operation, then immediately mini-tracheostomized, and then intermittent high-frequency jet ventilation during the daytime and pressure controlled ventilation during the nighttime were conducted. There were no differences in background factors between the two groups. The mean period of tracheal intubation was 8.8 days in the A group and 1.6 days in the B group. The mean period of stay in ICU was 13 days in the A group and 7 days in the B group. Both such periods were significantly shorter in the B group than in the A group. Respiratory control by early extubation and mini-tracheostomy after thoracic esophageal cancer operation is considered to have the following two benefits; 1. The method is performed safely, and makes sputum suction easier and assisted respiration conductible. 2. The method reduces the incidence of pulmonary complications and shortens the period of stay in ICU.

Using a large database of protein structure-structure alignments, we test a new method for distinguishing homologous and "analogous" structural neighbors. The homologous neighbors included in the test set show no detectable sequence similarity, but they may be well superimposed and show functional similarity or other evidence of evolutionary relationship. Analogous neighbors also show no sequence similarity and may be well superimposed, but they have different functions and their structural similarity may be the result of convergent evolution. Confirming results of other analyses, we find that remote homologs and analogs are not well distinguished by measures of pairwise structural similarity, including the percentage of identical residues and root-mean-square (RMS) superposition residual. We show, however, that with structure-structure alignments of analogous neighbors rarely superimpose the particular substructure that is shared among homologous neighbors. We call this characteristic substructure the homologous core structure (HCS), and we show that a cross-validated test for presence of the HCS correctly identifies 75% of remote homologs with a false-positive rate of 16% analogs, significantly better than discrimination by RMS or other measures of pairwise similarity. The HCS describes conservation of spatial structure within a protein family in much the way that a sequence motif describes sequence conservation. We suggest that it may be used in the same way, to identify homologous neighbors at greater evolutionary distance than is possible by pairwise comparison.

The characteristics of wavelength tunable femtosecond soliton pulse generation is investigated experimentally and numerically. The dependence of conversion efficiency and wavelength shift on the pump pulse is clarified.

The three dimensional structures for representatives of nearly half of all protein families are now available in public databases. Thus, no matter which protein one investigates, it is increasingly likely that the 3D structure of a homolog will be known and may reveal unsuspected structure-function relationships. The goal of Entrez's 3D-structure database is to make this information accessible and usable by molecular biologists (http://www.ncbi.nlm.nih.gov/Entrez). To this end Entrez provides two major analysis tools, a search engine based on sequence and structure 'neighboring' and an integrated visualization system for sequence and structure alignments. From a protein's sequence 'neighbors' one may rapidly identify other members of a protein family, including those where 3D structure is known. By comparing aligned sequences and/or structures in detail, using the visualization system, one may identify conserved features and perhaps infer functional properties. Here we describe how these analysis tools may be used to investigate the structure and function of newly discovered proteins, using the PTEN gene product as an example.

The phase noise and the amplitude noise of the amplitude modulated harmonically mode-locked Er-doped fiber ring laser have been investigated. They are smaller than those in conventional harmonically mode-locked Er-doped fiber ring lasers.