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The role that changes in DNA methylation and histone modifications have in human malignancies is poorly understood. p300 and CREB-binding protein (CBP), two distinct but highly homologous lysine acetyltransferases, are mutated in several cancers, suggesting their role as tumor suppressors. In the current study, we found that deletion of p300, but not CBP, markedly accelerated the leukemogenesis ofNup98-HoxD13(NHD13)transgenic mice, an animal model that phenotypically copies human myelodysplastic syndrome (MDS). p300 deletion restored the ability of NHD13expressing hematopoietic stem and progenitor cells (HSPCs) to self-renew in vitro, and to expand in vivo, with an increase in stem cell symmetric self-renewal divisions and a decrease in apoptosis. Furthermore, loss of p300, but not CBP, promoted cytokine signaling, including enhanced activation of the MAPK and JAK/STAT pathways in the HSPC compartment. Altogether, our data indicate that p300 has a pivotal role in blocking the transformation of MDS to acute myeloid leukemia, a role distinct from that of CBP.

DOCSPER—A Synthetic Lipid Fit for In Vivo ApplicationDOCSPER [1,3-Dioleoyloxy-2-(N5-carbamoyl-spermine)-propane] is a cationic amphiphile consisting of a hydrophobic 1,3 dioleylglycerol moiety and threefold positively charged spermine head group (1). We optimised the 5-step-synthesis of the lipospermine and after up-scaling we have obtained sufficient amounts to initiate preclinical investigations. DOCSPER was tested for its ability to transfect eukaryotic cells in vitro. It has proven to possess high transfection efficiency in comparison to commercially available liposomal transfection agents. Furthermore, DOCSPER was extensively tested in several in vivo studies (23). These studies revealed a high transfection efficiency, whereas very low toxicity levels were detected. Thus, the results clearly indicate that the cationic lipid DOCSPER is a reliable, low-risk system for broad applications in gene therapy.Groth D. et al. Int J Pharm 1998; 162:143–157.Nikol S. et al. Int J Angiol 2000; 9:87–95.Armeanu S. et al. Mol Ther 2000; 1(4):366–375.

Like other fields of medicine, robotics and mechanization might be introduced into endovascular coil embolization of intracranial aneurysms for effective treatment. We have already reported that coil insertion force could be smaller and more stable when the coil delivery wire is driven mechanically at a constant speed. Another background is the difficulty in synchronizing operators' minds and hands when two operators control the microcatheter and the coil respectively. We have therefore developed a mechanical coil insertion system enabling a single operator to insert coils at a fixed speed while controlling the microcatheter.Using our new system, the operator manipulated the microcatheter with both hands and drove the coil using foot switches simultaneously. A delivery wire force sensor previously reported was used concurrently, allowing the operator to detect excessive stress on the wire. In vitro coil embolization was performed using three methods: simple mechanical advance of the coil; simple mechanical advance of the coil with microcatheter control; and driving (forward and backward) of the coil using foot switches in addition to microcatheter control.The system worked without any problems, and did not interfere with any procedures. In experimental coil embolization, delivery wire control using the foot switches as well as microcatheter manipulation helped to achieve successful insertion of coils. This system could offer the possibility of developing safer and more efficient coil embolization. Although we aim at total mechanization and automation of procedures in the future, microcatheter manipulation and synchronized delivery wire control are still indispensable using this system.

Single crystal diamond micro-cutting tools, which are used for fabrication of precision micro-parts, have very sharp tool edges with radii in the range of tens of nanometers or even nanometers. Although there are many types of tools, the tools with a rounded nose are treated in this paper. They can be used for fabrication of very smooth and accurate surface with 3D micro structures such as micro-lens arrays, diffractive optical elements and so on. The machined quality is highly depending on the tool edges' states and it is desired to manage them. This paper presents a measuring instrument based on atomic force microscope (AFM), which is designed and constructed for on-machine measurement of the cutting edge profile. It is a combination of an AFM probe unit for 3D edge profile measurement and an alignment system with an optical sensor for aligning the probe-tip with the tool's edge top so that the measurement can be carried out in a short time. Measurement experiments of micro-tools with (nominal) nose radii of 8 m and 0.2 mm are summarized to show the performance of the instrument. And the nose radii were evaluated as 7.2 m and 0.188 mm, respectively.

This paper describes an atomic force microscope (AFM) based instrument for 3D edge profile measurement of single-point diamond cutting tools. The instrument is composed of an AFM unit and an optical sensor for alignment of the AFM probe tip (silicon cantilever) with the diamond cutting tool edge. In the optical sensor, a laser beam from a laser diode along the Y-axis is focused to generate a small beam spot with a micrometer-order diameter at the beam waist, and then received by a photo-detector (photodiode). The tool edge top and the AFM probe tip are brought to the center of the beam waist in the XZ-plane through monitoring the variation of the photodiode output, respectively. Consequently, the AFM tip can be aligned with the tool edge top. Alignment experiments and 3D edge profile measurements of a round-nose type single-point diamond tool are carried out.

AbstractIn the field-reversed configuration (FRC) plasma, rotational instability with toroidal mode number n= 2 is known as a sole destructive MHD instability. Though an ideal FRC is unstable for higher n interchange instabilities, those have never been observed in past experiments. As a possible stabilized mechanism, self-generated toroidal rotation and its shear have been investigated experimentally.

Rheo-casting of 4045 aluminum alloy bar was operated using an unequal diameter twin roll caster and a cooling slope. The semisolid slurry was made by the cooling slope. The thickness of the bar was 12 mm and width was 100mm. The casting speed was 1m/min. Quantity of the melt poured on the cooling slope was important to make sound bar. The primary crystal became spherical, and its size was about 40μm.

A neutral beam injection (NBI) into a field-reversed configuration (FRC) is simulated by the electron-fluid and ion-particle method. The neutral beam particle is injected tangentially to the field-null circle, and the simulation is done on a two-dimensional cross section of the FRC. An initial equilibrium state is obtained from the Grad–Shafranov equation including the beam current term. It is found that the start of radial ion flow is delayed by the NBI.

Only a prototype laryngeal tube has been compared with the classic LMA for brief periods of anaesthesia. We compared the new laryngeal tube (which had several improvements in design) with the classic LMA.

Background.There are two forms of Macintosh laryngoscope blade. Compared with the standard blade, the English blade is longer, its curve is more continuous across the entire length of the blade, the flange of the blade continues much closer to the blade tip, and the height of the flange is shorter.

We have compared the laryngeal tube and laryngeal mask in 22 patients for the success rate of insertion, gas leak pressure and the incidence of gastric insufflation.

Abstract.:  A male patient initially diagnosed with acute lymphoblastic leukemia at age 9 years received chemotherapy (total body irradiation, 12 Gy) followed by allogeneic bone marrow transplantation. Since then, he had been in complete remission. Three years after the bone marrow transplantation, he complained of increasing pain in the right knee. Radiological and histological examinations led to a diagnosis of conventional osteosarcoma. We performed intensive chemotherapy and wide local excision of the osteosarcoma. Intensive chemotherapy was accomplished as planned, although recovery from myelosuppression was delayed during some cycles. Polymerase chain reaction-single-strand conformation polymorphism analysis revealed a p53 gene mutation in exon 7 in the tumor cells, but not in skin or blood cells. This is an extremely rare case of osteosarcoma after bone marrow transplantation.

AbstractTwenty-five hips in 19 rheumatoid arthritis (RA) patients with protrusio acetabuli were followed up, both clinically and radiographically, for more than 9 years after total hip arthroplasty (THA), that was performed with a bone graft to reinforce the medial acetabular wall. Radiographs were taken preoperatively and every 6 months postoperatively. Clinical assessments of pain, gait, and range of motion of the hips were obtained preoperatively and every year postoperatively using the Japanese Orthopaedic Association hip-scoring system. Radiographs showed that bony union had occurred in all cases. Six acetabular components were loose, but no femoral components became loose during the 9-year period. The clinical evaluation showed that relief of pain was very significant. The range of motion of the hip joints also improved from 12 points to 16.6 points after 9 years. Walking ability improved, but is becoming worse as time goes by. The radiographic results were compared with the results of a THA group with RA that had not had a bone graft. The rate of loosening of the THA without a bone graft was significantly higher than that of THA with a bone graft. We concluded that bone grafting for protrusio acetabuli was a very useful procedure.

Abstract: Background. The intrarenal localization of metallothionein (MT) expression has not been studied in detail. Therefore, we investigated the effects of acute exposure to cadmium on the renal expression of MT. Methods. MT mRNA expression was estimated quantitatively at 2, 6, 24, and 72 h after the injection of cadmium (1 mg/kg) to the intraperitoneum of male Sprague-Dawley rats. The site of cadmium-induced MT mRNA expression was determined, by the reverse transcription polymerase chain reaction (RT-PCR), using microdissected tubules. Immunoblotting and immunohistochemistry were also performed, to determine the localization of MT protein. Results. The mRNAs of MT-I and MT-II were significantly upregulated at 2 h and peaked at 24 and 6 h, respectively. RT-PCR revealed the expression of cadmium-induced MT mRNA mainly in the proximal tubules. Immunoblotting detected a clear 14-kDa band corresponding to MT protein, and immunohistochemistry showed abundant MT protein in the renal cortices of cadmium-treated rats. Conclusions. Our findings suggest that the proximal tubules are responsible for the cadmium-induced renal expression of MT mRNA and protein.

Crack propagation tests were performed on an amorphous polymer, poly(methyl methacrylate), to investigate fatigue crack propagation mechanisms. A scanning laser microscope with a newly developed tensile testing machine was used to observe in situ crack propagation in compact-type specimens. A crack usually propagated within the craze located at the crack tip under both static and cyclic loading conditions. When a crack stably propagated into the craze under static loading conditions, bright bands composed of the broken craze were observed at the edges along the crack wakes. However, there were successive ridges and valleys in place of bright bands along the crack wakes under cyclic loading conditions. When stable fatigue cracks were propagated at the loading half-cycle in each cycle, new craze fragments appeared that were similar to the bright bands under static loading. However, the thickness of these fragments decreased in the following loading cycle, and a new valley was formed. This suggested that the valleys were formed by the contact between the fracture surfaces near the crack tip during unloading. Fatigue crack propagation is thought to be due to fibrils weakened by crack closure between fracture surfaces. © 2001 John Wiley & Sons, Inc. J Polym Sci Part B: Polym Phys 39: 3103–3113, 2001

Peripheral blood progenitor cells are collected effectively by leukapheresis of a large volume of peripheral blood. However, protection must be taken for patients or donors from hypocalcemia due to continuous infusion of citric acid. We found a tendency for hypocalcemic symptoms in patients or donors to occur more often on the second day than the first day of the sequential 2 days of leukapheresis. The doses of calcium gluconate supplement and the acid citrate dextrose-A solution administration significantly increased on the second day compared to that of the first day. The blood levels of c-terminal parathormone (PTH), phosphorus, and alkaline phosphatase did not show remarkably abnormal change. However, urine calcium excretion just after leukapheresis was higher than in the period before or after leukapheresis compared to the phosphorus or creatinine excretion. These findings indicate that the cause of a higher tendency to hypocalcemic symptoms on the second day of the sequential 2 days of leukapheresis is due to the higher metabolism of calcium being excreted in the urine during leukapheresis.

Abstract: Background. Polymorphism of the collagen type Iα1 gene (COLIA1) has been shown to be involved in bone density in the general population. However, the significance of this polymorphism has not been examined in patients with chronic renal failure. Methods. We first determined the COLIA1 Bal I restriction fragment length polymorphism (RFLP) at the Sp 1 binding site, and, next, we measured biochemical bone markers and the bone mineral density (BMD) of the lumbar spine (L2–L4) and radius (distal one-third) in 181 hemodialyzed patients. Results. Distributions of the Bal I RFLP were: SS, 62%; Ss, 32%; and ss, 6%. The Z scores for the BMD and for the serum levels of Ca, P, alkaline phosphatase, intact-parathyroid hormone (PTH), osteocalcin, and tartrate-resistant acid phosphatase were not significantly different among the three genotype groups. The slope of the regression line in the age-BMD relationship, a measure of the rate of BMD decrease, was similar among the three genotypes in men, but was significantly (P < 0.05) larger in the ss group than in the SS and Ss groups in women. Conclusions. The COLIA1 polymorphism may be associated with the bone loss seen in female patients on hemodialysis.

OBJECTIVE: To determine whether endorphin (END) and enkephalin (ENK) modulate excessive synovial cell functions in patients with rheumatoid arthritis (RA). METHODS: Effects of leucine-enkephalin (leu-ENK), methionine-enkephalin (met-ENK), and beta-endorphin (END) on proinflammatory cytokine and matrix metalloproteinase (MMP) production by RA synovial cells were analyzed by immunoblotting, and their mRNA expression by reverse transcription-polymerase chain reaction (RT-PCR) using limiting dilution of complementary DNA. Expression of opioid receptors on RA synovial cells was assessed by immunohistochemical staining, radioreceptor assay, and RT-PCR. RESULTS: Leu-ENK, met-ENK, and END inhibited tumor necrosis factor-alpha and interleukin 1beta production at the level of mRNA expression. ENK and END inhibited MMP-9 production and its enzymatic activity by RA synovial cells. The mu-subtype opioid receptor was expressed in the RA synovial lining and sublining cells. Radioreceptor assay suggested expression of high affinity receptor for END on RA synovial cells. The mu-subtype opioid receptor-specific antagonist, naloxone, reversed the inhibitory effect of the opioid peptides. The opioid peptides inhibited nuclear translocation and phosphorylation of the transcription factor, cyclic AMP responsive element binding protein (CREB) in RA synovial cells. CONCLUSION: Leu-ENK, met-ENK, and END inhibited excessive RA synovial cell functions in vitro. The opioid hormones may have not only antinociceptive action, but also antiinflammatory effects on synovitis itself in RA.

We investigated whether low‐dose erythromycin (200 mg) given as an oral premedicant altered the residual gastric volume and its acidity in fasted patients at induction of anaesthesia in a single‐blinded study. Sixty patients were allocated randomly to receive either an erythromycin tablet (200 mg) or nothing with 10 ml water 3 h before induction of anaesthesia, and another 60 patients 1 h before induction of anaesthesia. Oral erythromycin significantly reduced residual gastric volume when it was given approximately 3 h (P<0.05; 95% CI for median difference: 0.1–17 ml) or 1 h (P<0.0005; 95% CI for median difference: 6–24 ml) before induction of anaesthesia. Erythromycin significantly reduced gastric acidity when it was given 1 h before induction of anaesthesia (P<0.02; 95% CI for median pH difference: 0.1–1.7). In contrast, when given 3 h before induction of anaesthesia, erythromycin did not significantly alter acidity.Br J Anaesth 2000; 85: 861–4

We studied 70 patients to see if cricoid pressure applied after insertion of the laryngeal mask altered the success rate of tracheal intubation through the mask. After induction of anaesthesia and neuromuscular blockade, patients were randomly allocated to have either cricoid pressure (Group C) or sham pressure (Group S). The view of the glottis through the laryngeal mask was assessed before and after the test pressure, and tracheal intubation through the mask was attempted using a fibreoptic bronchoscope. The test pressure did not alter the view of the glottis in any patient in group S, whereas it narrowed the glottic aperture in 16 out of 35 patients in group C. The fibrescope was inserted into the trachea in all patients in group S and in 25 patients in group C. The success rate of tracheal intubation in group S (31 patients) was significantly higher than in group C (21 patients, P << 0.001; 95% CI for difference: 9-48%). The time for insertion of the fibrescope in group S (median (95% CI): 12 (11-12) s) was significantly faster than in group C (16 (14-17) s, P << 0.001; 95% CI for difference: 3-6 s), and the time for tracheal intubation in group S (16 (15-18) s) was significantly faster than in group C (22 (19-24) s, P < 0.0005; 95% CI for difference: 3-7 s). Cricoid pressure after insertion of the laryngeal mask makes tracheal intubation through the mask significantly more difficult.

Frequencies of vaccine-responsive T-lymphocyte precursors in peripheral blood mononuclear cells (PBMC) prior to and after administration of peptide-based vaccines in patients with cancer can be measured by limiting-dilution assays (LDA) or by ELISPOT assays. We have used a modified version of the ELISPOT assay to monitor changes in the frequency of gamma interferon (IFN-gamma)-producing T cells in a population of lymphocytes responding to a relevant peptide or a nonspecific stimulator, such as phorbol myristate acetate-ionomycin. Prior to its use for monitoring of patient samples, the assay was validated and found to be comparable to the LDA performed in parallel, using tumor-reactive cytolytic T-lymphocyte (CTL) lines. The sensitivity of the ELISPOT assay was found to be 1/100,000 cells, with an interassay coefficient of variation of 15%, indicating that it could be reliably used for monitoring of changes in the frequency of IFN-gamma-secreting responder cells in noncultured or cultured lymphocyte populations. To establish that the assay is able to detect the T-cell precursor cells responsive to the vaccine, we used CD8(+) T-cell populations positively selected from PBMC of HLA-A2(+) patients with metastatic melanoma, who were treated with dendritic cell-based vaccines containing gp100, MELAN-A/MART-1, tyrosinase, and influenza virus matrix peptides. The frequency of peptide-specific responder T cells ranged from 0 to 1/2,600 before vaccination and increased by at least 1 log unit after vaccination in two patients, one of whom had a clinical response to the vaccine. However, no increases in the frequency of peptide-responsive T cells were observed in noncultured PBMC or PBMC cultured in the presence of the relevant peptides after the melanoma patients enrolled in another trial were treated with the intramuscular peptide vaccine plus MF59 adjuvant. Thus, while the ELISPOT assay was found to be readily applicable to assessments of frequencies of CTL precursors of established CTL lines and ex vivo-amplified PBMC, its usefulness for monitoring of fresh PBMC in patients with cancer was limited. In many of these patients antitumor effector T cells are present at frequencies of lower than 1/100,000 in the peripheral circulation. Serial monitoring of such patients may require prior ex vivo amplification of specific precursor cells.

Thylakoidal proteins of plant chloroplasts are transported to thylakoids via several different pathways, including the DeltapH-dependent and the Sec-dependent pathways. In this study, we asked if these two pathways utilize a common translocation pore. A fusion protein consisting of a 23-kDa subunit of the oxygen evolving complex and Escherichia coli biotin carboxyl carrier protein was biotinylated in E. coli cells and purified. When incubated with isolated pea thylakoids in the absence of avidin, the purified fusion protein was imported into the thylakoids via the DeltapH-dependent pathway. However in the presence of avidin, the fusion protein became lodged in the thylakoid membranes, with its N terminus reaching the thylakoidal lumen, while its C-terminal segment complexed with avidin exposed on the thylakoidal surface. The translocation intermediate of the fusion protein inhibited the import of authentic 23-kDa subunit, suggesting that it occupies a putative translocation pore for the DeltapH-dependent pathway. However the intermediate did not block import of the 33-kDa subunit of the oxygen evolving complex, which is a substrate for the Sec-dependent pathway. These results provide evidence against the possibility of a common translocation pore shared by the Sec-dependent pathway and the DeltapH-dependent pathway.