Search

To clarify an inexpensive method for constructing a software-defined network (SDN) environment, we propose a router control function for non-SDN-supported routers using IoT devices possessing a common user interface based on OpenFlow. An experimental demonstration using the proposed function is shown.

Objectives The aim of this study was to evaluate an influence of post-processing scatter correction in portable abdominal radiography using a low ratio anti-scatter grid (grid). Methods To assess tube voltage on portable abdominal radiography, a burger phantom was used to measure for inverse of image quality figure (IQFinv). For evaluation of the influence on using or not the grid, IQFinv were measured. Abdominal phantom radiographies were assessed subjectively, in random order, by six radiologic technologists. The radiographies were performed without scatter correction [IG (-)] and with scatter correction at equivalent for grid ratio 6 [IG (6)] and 8 [IG (8)]. Results There was no significant decrease in IQFinv with 75 and 80 kV in comparison of 70 kV. Even processing scatter correction, IQFinv with using the grid was significantly higher than that without using the grid. The ability to detect nasogastric tube and stomach gas were significantly better in the scatter correction. Deviation index for IG (6) and IG (8) were significantly lower than that of IG (-). Discussion Portable abdominal radiographies will be improved image quality by utilizing scatter correction, although, it is necessary to consider the scatter correction processing as this may significant decrease deviation index in the practical situation. Conclusion The post-processing scatter correction should be useful for detection nasogastric tube and stomach gas in portable abdominal radiography.

Disaggregation breaks conventional closed systems into components connected by open interfaces. This paper shows the experimental demonstration of service provisioning and partial replacement of network OS in a disaggregated open packet and optical converged network based on open interfaces and open source software.

OpenFlow-based remote control system for the optical switch in AWG-STAR with a loopback function using IoT device is proposed as a low-cost solution to extend the scope of a software-defined network to physical layer.

Maternal antibodies against human platelet antigen (HPA) and/or human leukocyte antigen (HLA) cause fetal and neonatal alloimmune thrombocytopenia (FNAIT) in 0.09-0.15% of live births. Severe cases account for 5-31% and the frequency of multiple kinds of alloantibodies is 6.9-9% of FNAIT. We present a case of severe FNAIT associated with anti-HPA-5b, anti-HLA-A31, and anti-HLA-B55 antibodies, successfully treated with immunoglobulin and platelet transfusion. The anti-HLA-B55 antibody was detected in the newborn's serum, but disappeared on the 20th day, which was followed by an increase of the platelet count. These findings suggested the potential involvement of an anti-HLA antibody in the pathogenesis of FNAIT.

Objectives The aim of this study was to evaluate the effectiveness of scatter correction in the portable chest radiography. Methods Digital radiographies were performed without anti-scatter grid (grid), with the scatter correction and with the grid ratio of 3 : 1 in this study. The scatter fraction and the detectability of low contrast signals were measured using the four acrylic phantoms of different thicknesses. The chest phantom radiographs were assessed subjectively, in random order, by six radiologic technologists. Results The scatter fraction was higher in the no-grid technique, and was lower for the grid technique. The detectability of low contrast signals did not significantly differ between the scatter correction and the grid technique (p>0.05). The area under the receiver operating characteristic curve for the grid technique was higher than that for the scatter correction technique (0.888 vs. 0.855), although no significant difference was found between the grid and the scatter correction technique (p> 0.05). The ability to detect the nasogastric tube was significantly better in the grid technique (p Discussion In the scatter correction technique, the ability of scatter removal increased as the scatter fraction increased. The scatter correction technique was unnecessary to extremely accurate alignment. In addition, patient dose can be reduced by the scatter correction technique. Conclusions It seemed to be effective for the scatter correction in the portable chest radiography.

We have developed a new method to determine the N-terminal amino acid sequences of proteins, regardless of whether their N-termini are modified. This method consists of the following five steps: (1) reduction, S-alkylation and guanidination for targeted proteins; (2) coupling of sulfo-NHS-SS-biotin to N(alpha)-amino groups of proteins; (3) digestion of the modified proteins by an appropriate protease followed by oxidation with performic acid; (4) specific isolation of N-terminal peptides from digests using DITC resins; (5) de novo sequence analysis of the N-terminal peptides by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) using the CAF (chemically assisted fragmentation) method or tandem mass spectrometric (MS/MS) analysis according to unblocked or blocked peptides, respectively. By employing DITC resins instead of avidin resins used in our previous method (Yamaguchi et al., Rapid Commun. Mass Spectrom. 2007; 21: 3329), it has been possible to isolate selectively N-terminal peptides from proteins regardless of modification of N-terminal amino acids. Here we propose a universal method for N-terminal sequence analysis of proteins.

Use of a bis(terpyridine)ruthenium(II) derivative as an N-terminal labeling reagent resulted in the simultaneous detection and individual determination of all the N-terminal fragments of the proteins in a mixture without requiring any separation. All of the N-termini of the guanidinated proteins were labeled selectively by the ruthenium complex (Ru-CO-labeling). After chymotryptic digestion, the fragments were analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) and post-source decay (PSD). TheRu-CO moiety exclusively enhanced N-terminal fragment ions in mass spectra and enabled easy N-terminal sequencing. In a mixture containing three different proteins (lysozyme, ubiquitin, and insulin), all of the N-terminal fragment ions labeled with the ruthenium complex were found to produce uniformly intense peaks without the detection of the other unlabeled fragments. The N-terminal sequences of these ions were determined individually by PSD analysis. Application to unknown proteins from Thermus thermophilus HB8 with two-dimensional electrophoretic separation resulted in the successful determination of the N-terminal sequence and easy identification of the target protein.

A new method to determine N-terminal amino acid sequences of multiple proteins at low pmol level by a parallel processing has been developed. The method contains the following five steps: (1) reduction, S-alkylation and guanidination for targeted proteins; (2) coupling with sulfosucccimidyl-2-(biotinamido)ethyl-1,3-dithiopropionate(sulfo-NHS-SS-biotin) to N(alpha)-amino groups of proteins; (3) digestion of the modified proteins by an appropriate protease; (4) specific isolation of N-terminal fragments of proteins by affinity capture using the biotin-avidin system; (5) de novo sequence analysis of peptides by MALDI-TOF-/MALDI-TOF-PSD mass spectrometry with effective utilization of the CAF (chemically assisted fragmentation) method.1 This method is also effective for N-terminal sequencing of each protein in a mixture of several proteins, and for sequencing components of a multiprotein complex. It is expected to become an essential proteomics tool for identifying proteins, especially when used in combination with a C-terminal sequencing method.

A full-length cDNA clone encoding osteocalcin from the bullfrog, Rana catesbeiana (bone Gla-protein, BGP) has been isolated, and the complete coding sequence for the 100-amino-acid pre-pro-osteocalcin protein was determined. The amino acid sequence of Rana catesbeiana osteocalcin, especially the mature 49-amino acid sequence, is closer to the mammalian than to the fish, Sparus osteocalcin. Rana mature osteocalcin has a similarity of 67% with human or 59% with rat osteocalcin, and only 42% with fish mature osteocalcin. The 51-amino-acid pre-pro-peptide contains the expected hydrophobic leader sequence and the dibasic Arg-Arg sequence preceding the NH 2 -terminal Ser of the mature 49-amino-acid Rana osteocalcin. The pro-peptide sequence also contains the expected motif of polar and hydrophobic residues, which targets vitamin K-dependent gamma-carboxylation of three specific Glu residues at positions 17, 21, and 24 in the mature protein. At the native protein expression levels, extraction from Rana cortical bone in the presence of protease inhibitor cocktail resulted in the isolation of two distinct forms of osteocalcin, P-1 and P-2, with a 3:2 distribution. Using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) and amino acid sequence analysis of the N-terminal domain, we confirmed that P-1 is the intact 49-residue osteocalcin with N-terminal SNLRNAVFG·, and that P-2 lacks four amino acids from the N-terminus, (NAVFG·). These results demonstrate the existence of a form of osteocalcin lacking four N-terminal amino acids in Ran a bone, and that mature Rana osteocalcins remained highly conserved in their molecular evolution, especially with respect to the conservation of the C-terminal domain (residues 14–49).

We have developed a novel method for enhancing the response of a peptide in matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) by activating the C-terminal carboxyl group through an oxazolone with which is coupled an amine containing a functional group to help ionize the peptide. The reactions consist of dehydration with acetic anhydride to give an oxazolone, followed by aminolysis with an appropriate amino acid derivative such as arginine methyl ester. The MALDI signal of Ac-Tyr-Gly-Gly-Phe-Leu-Arg-OMe, thus converted from leucine-enkephalin, was detected while completely excluding the responses of arginine-deficient peptides coexisting in the reaction mixture. Some less intense peaks corresponding to a few sequential degradation products, also terminated with the arginine derivative, were also observed. The side-chain groups potentially that are reactive were conveniently protected by acetylation simultaneous with the C-terminal activation, and those that remained unprotected were reduced to virtually negligible proportions when the reaction was conducted in a peptide solution of concentration less than 1 mM. The greatly increased responses of such arginine-terminated peptides could possibly be exploited to discern the C-terminal tryptic peptide of a protein that is otherwise almost insensitive to MALDI-MS in general. The simplicity of the post-source decay spectrum of enkephalin derivatized by arginine methyl ester characteristically accentuated z- and b-type ions, and this should facilitate sequencing of such derivatized peptides. Remaining problems with practical applications of this approach are discussed.

This paper proposes a new control scheme for a coordinated motion and force control of multi-fingered robot hand. Proposed control scheme can achieve simultaneous control of the position of a target object and a so-called ‘internal force’ which does not affect the motion of the object. Based on ‘knetostatic filtering method’ which eliminates a element of internal force from joint angle feedback signal, proposed controller can achieve internal force regulation for the fingers which have kinematical redundancy. Computer simulation results and experimental results illustrates the effectiveness of proposed scheme.

The phase separation temperatures of dilute solutions of 4He in liquid 3He in the Fermi-liquid region have been measured by detecting the 4He film formation sensitively. From the obtained solubility curve, the effective mass of 4He atom and the difference in binding energy of 4He in liquid 4He and in liquid 3He are proposed.

Boiling behavior of liquid 3He below 1 K was visualized as shadowgraph image. A light source and a video camera (or still camera) were arranged at room temperature. The light was guided to the 3He cell by an optical fiber, and the shadowgraph image was transferred to the camera by an image fiber. The 3He bubble formed on the heated copper surface was spheroid-like due to the low surface tension. The size at departure from the surface and the relative bubble formation rate were measured as a function of heat flux. The previously obtained heat transfer curve could be explained qualitatively by this boiling behavior.

We propose a novel IP over AWG-STAR network with dynamically relocatable wavelength paths via communication node-side control. Experimental results demonstrate that the transmission capacity between communication nodes can be dynamically increased from 1 Gbps to 3 Gbps via adding looped-back wavelength paths to the internode in the proposed IP over AWG-STAR network we constructed experimentally.

Effects of “eddy viscosity” on superfluid turbulence associated with the vortex tangle are discussed in terms of the interpretation of pressure difference in various types of capillary flow and also in regard to the velocity profiles of normal- and superfluid over the capillary cross section. The eddy viscosity coefficient and the boundary value of superfluid velocity on the capillary wall are treated as parameters that are determined by fitting to the observed data of pressure differences in the pure superflow and thermal counter flow. Using these parameters, it is shown that in the well-developed turbulent state the relative velocity between the normal fluid and the superfluid shows almost uniform profile over the cross section of the capillary except very near the boundary wall. This situation partially supports recent successful analysis of temperature difference, which relied on the dynamical scaling theory in the assumed homogeneous velocity fields and distributions for the vortex tangle.

The cyanobacterium Synechocystis sp. strain PCC6803 is an interesting model organism for preoteome study because it is a photosynthetic procaryote and its genomic sequence has already been determined at our institute. We thus initiated characterization of this organism from a proteomic viewpoint by exploiting two-dimensional (2-D) gel electrophoresis coupled with N-terminal protein sequencing. In a previous study, we linked 130 protein spots on two dimensional gels with the genes that encoded them. As an extension of the previous study, the number of protein spots linked to their corresponding genes was increased to 227 in this study by separately analyzing cyanobacterial proteins in four different fractions (soluble, insoluble, thylakoid membrane, and secretory protein fractions). The resultant updated 2-D protein-gene linkage database, named Cyano2Dbase, will serve as an indispensable tool in future cyanobacterial proteomic studies. From the data compiled in the Cyano2Dbase, we can extract many items of information concerning translation, posttranslational processing including characteristics of cyanobacterial signal sequences and modification of cyanobacterial proteins. The Cyano2Dbase is available to the public through the World Wide Web (http://www.kazusa.or.jp/tech/sazuka/cyano/pr oteome.html).

Fly ash from a municipal solid waste incinerator is regulated under the hazardous waste regulation “Waste under Special Control”, according to the Amendment of the Waste Disposal and Public Cleaning Law in Japan, because it contains high concentrations of heavy metals which are available for leaching.To evaluate the toxicity of fly ash, at first, a fly ash leachate was prepared according to the Japanese standard leaching procedure. The chemical analysis of the leachate showed that one of the possibly most toxic substances was cadmium. The toxicity of the leachate and cadmium was determined by algal assay and Daphnia acute toxicity test. The results showed that the leachate was about 10 times more toxic to the growth of algae, and 20 to 30 times more toxic to the survival of Daphnia than expected from its cadmium concentration.The toxicity interaction between cadmium and the other constituents in the leachate was also examined. The toxicity of cadmium showed additive effect with the other constituents in the leachate in algal assay. In Daphnia test, however, cadmium showed antagonistic effect.

The viscosity of liquid 4He has been measured by means of a high-Q torsional oscillator at temperatures between 1.5K and 2.7K under saturated vapor pressure. The viscosity data derived from the resonant frequency difference between the empty cell and the cell filled with liquid 4He show satisfactory agreement with the results obtained by Welber. When the velocity of cell-wall exceeded the critical velocity below the superfluid transition temperature, the decrease of the resonant frequency caused by the growth of the quantized-vortex was observed. In the case of a cell with rough wall, the critical velocity was low compared with a cell with smooth wall. The rate of the growth of vortex line length also depends on the wall roughness.

溶出試験は廃棄物の有害性評価の一指標として広く用いられている。しかし, その方法が多様であるため, 結果の科学的な解釈が難しい, 異なる方法による結果の比較ができないなどの問題を抱えている。これを解決する手段として最大溶出可能量を測定することを目的とした溶出試験が注目されている。本研究では, 都市ごみ焼却飛灰中に含まれる重金属を対象として攪拌強度, 固液比, 接触時間, pHの溶出操作条件をパラメータとした溶出実験を行い, それぞれの条件下で溶出する金属量を調べることによって最大溶出可能量を調べるための溶出操作条件について検討した。その結果, カドミウム, 銅, 亜鉛およびマンガンについては最大溶出可能量を把握する溶出条件を設定できた。しかし, この条件では鉄の溶出量は接触時間の影響を, 鉛の溶出量は固液比および酸の種類による影響を受けることが明らかとなった。このように成分によって溶出の制限となる操作因子が異なるため, 最大溶出可能量を把握するための溶出試験方法を決めるには, 対象成分に応じて適当な溶出操作条件を検討する必要があることが示された。

Two HLA-B*3501 binding self-peptides, LPFDFTPGY (37F) and LPGPKFLQY (28H), were isolated from HLA-B*3501 molecules expressed by cultured human B lymphoid cells. Both sequences were consistent with previously reported motifs of HLA-B*3501 binding peptides which carry proline at position 2 and tyrosine at position 9 as anchor residues. Direct binding of these peptides to HLA-B*3501 molecules was quantitated by flow cytometry analysis of RMA-S cells. transfected with the HLA-B*3501 gene (RMA-S-B*3501). Both 37F and 28H peptides bound effectively to HLA-B*3501 molecules. Substitution of amino acids at position 2 and/or 9 of HLA-B*3501 binding peptides markedly reduced their binding to HLA-B*3501 molecules. These results indicate that two anchor residues, proline at position 2 and tyrosine at position 9 are critical in binding of peptides to HLA-B*3501 molecules. Insertion of up to four glycine residues at position 8 of the peptide 37F did not affect its binding affinity to HLA*3501 molecules. These results indicate that long peptides can effectively bind to HLA class I molecules provided that anchor residues are conserved.

Iodine deficiency in pregnant and lactating women results in serious damage to their fetuses, newborns, and weaning infants. The effect of dietary iodine intake on maternal and infantile thyroid function has not been well studied in iodine-sufficient areas, and there are few data on appropriate gestational age-specific reference ranges for urinary iodine excretion during pregnancy and lactation.The aim of the study was to characterize the gestational change of urinary iodine excretion in Japanese women and to assess the effects of iodine status on thyroid function in mother and infant.A total of 934 Japanese women and their 722 newborn infants were enrolled in the study. Iodine and creatinine concentrations were determined in spot urine samples in the three trimesters of pregnancy and the postpartum period at 34.0 d after delivery. Serum thyroperoxidase antibody and thyroglobulin antibody, TSH, and free T(4) were measured in each trimester, and neonatal TSH was measured on postnatal d 4.The overall median urinary iodine concentration (UIC) during pregnancy was 219.0 μg/liter, higher than that in postpartum women (135.0 μg/liter). The prevalence of pregnant women with low UIC less than 100 μg/liter or high UIC greater than 500 μg/liter was 16.1 and 22.2%, respectively. Urinary iodine excretion decreased from 221.0 μg/liter in the first trimester to 208.0 μg/liter in the second trimester to 193.0 μg/liter in the third trimester, and then remained at 135.0 μg/liter postpartum. [corrected]. The maternal UIC correlated positively with serum TSH during pregnancy. There was no significant difference in UIC between subjects with positive thyroid autoantibodies and those with negative antibodies.Iodine intake assessed by UIC in Japanese pregnant women is regarded as sufficient and not excessive according to World Health Organization criteria. Although the data are local, our results provide additional information on the reference range for UIC throughout gestation in iodine-sufficient areas.

Prediction of the low Mach number jet noise ejected from rectangular nozzle with high aspect ratio is described. Firstly measurement of the jet noise was conducted in semi-anechoic wind tunnel at the low Mach number flow condition. We were found that the sound power of the jet obeyed 6-th power of jet velocity. This means the jet noise is resulted not from quadrupole distribution in the shear flow of jet, but from dipole distribution on the surface of the exit of the nozzle. The model of vortex sound is applied as the sound generation mechanism at numerical simulation. The sound emission from the vortices in the shear flow is modified with the compact Green’s function representing the scattering effect from the surface of the exit of the nozzle in the lower frequency range. It is also modified with the non-compact Green’s function in the higher frequency range. Lastly calculated sound spectra are compared with measured spectra. The comparison will prove effectiveness of this modeling.

Polymerization of MMA with R3P-R3′Al in toluene at low temperatures was reinvestigated and was found to give highly syndiotactic PMMA. Among the initiators examined Ph3P-Et3Al gave the PMMA with highest syndiotacticity and the polymerization at −93°C yielded PMMA with rr content of 95%. PMMAs prepared with Et3P-Et3Al and Ph3P-Et3Al contained the structural unit comprised of the phosphine components at the chain end as evidenced by 31P and 1H NMR analyses. The results clearly indicate that R3P takes an important role in the initiation process, though R3P alone does not polymerize MMA. Methacrylic acid esters of primary and secondary alcohols were also polymerized by these initiators to give highly syndiotactic polymers. Block and random copolymers of MMA with several methacrylates were also obtained. The effect of syndiotacticity on the glass transition temperature (Tg) of PMMA was examined and Tg for 100% syndiotactic PMMA with infinite molecular weight was estimated to be 141°C.

We studied the use of hepatic parenchyma as a recipient site for pancreatic fragment transplantation. Endocrine and exocrine pancreatic functions were evaluated following pancreatic autotransplantation in 26 mongrel dogs that had undergone total pancreatectomy. The endocrine function of the pancreatic tissue transplanted to hepatic parenchyma was significantly inferior to that of normal controls. Cholecystic bile amylase concentrations were markedly elevated in six dogs that had been implanted with pancreatic fragments in their hepatic parenchyma and had survived more than 2 months. Also, choledochal bile amylase concentrations increased significantly following pancreozymin-secretion (PS) injection. In contrast, cholecystic bile amylase concentrations in normal dogs were low and choledochal bile amylase concentrations did not respond to a PS load. Histological examination of pancreatic autografts in hepatic parenchyma revealed marked proliferation of exocrine tissue with abundant zymogen granules and reconstruction of pancreatic lobules with a few islets.

A facile method for routine sequence analysis of cysteine and cystine containing peptides or proteins is described. An automated gas-phase protein sequencer, Shimadzu PSQ-1 System, consisting of an Edman-reaction unit and an isocratic separation mode for the phenylthiohydantoin amino acid analysis with equivalent sensitivity and resolution to that of the current gradient mode, allows easy determination of the intact second half cystine, of which phenylthiohydantoin derivatives appear as a dithiothreitol adduct of dehydroalanine, cysteine and S- pyridylethylated cysteine. The optimal sequencing protocol for in situ pyridylethylation was established. This protocol gives an N-terminal amino acid sequence with half cystines, which are automatically converted to the S-pyridylcthyl derivative in the sequencer. Sequence analyses both intact and S-pyridyiethylated cysteine can identify a disulfide form if only one disulfide linkage exists in the sample. Sequence analysis of synthetic peptides containing ...

Three sites were evaluated for potential pancreatic fragment autotransplantation. Both endocrine and exocrine functions were evaluated following autotransplantation into splenic pulp, portal vein, or hepatic parenchyma in 44 pancreatectomized dogs. Cholecystic bile amylase concentrations in the hepatic parenchyma group surviving more than 2 months were elevated significantly, and choledochal bile amylase concentrations increased markedly following pancreozymin-secretin injection. In contrast, bile amylase concentrations in dogs with intrasplenic or intraportal implants were low and did not respond to PS injection. Histologically pancreatic autografts in hepatic parenchyma revealed marked proliferation of exocrine tissue with abundant zymogen granules and reconstruction of the acinar lobules with a few islets. These acinar cells in the hepatic parenchyma were ultrastructurally normal. Transplant endocrine function, estimated by K values, was significantly better after splenic pulp and portal vein than after a hepatic parenchyma implantation, but no group improved during 1-year follow-up. Glucose-stimulated initial insulin responses were abnormally low in all recipients. Islet B cells lacked mature insulin granules, such as seen in normal resting B cells. This ultrastructural finding implies a persistent demand on the B cells and may explain the spontaneous recurrence of hyperglycemia and the diminished initial insulin response to a glucose load. This study indicates that euglycemic recipients of pancreatic fragment autotransplantation remain unstable and prediabetic.

We have measured the heat transfer in superfluid helium in a glass capillary tube. The vortex viscosity calculated from pressure difference across the tube was linear to L_0^ in the range 15ddd and L_0d^2,we could evaluate the vortex-wall interaction parameter ξ' and ξ.

A simple and efficient method for C-terminal sequencing of proteins has long been pursued because it would provide substantial information for identifying the covalent structure, including post-translational modifications. However, there are still significant impediments to both direct sequencing from C termini of proteins and specific isolation of C-terminal peptides from proteins. We describe here a highly successful, de novo C-terminal sequencing method of proteins by employing succinimidyloxycarbonylmethyl tris (2,4,6-trimethoxyphenyl) phosphonium bromide and mass spectrometry.

In proteomics, MS plays an essential role in identifying and quantifying proteins. To characterize mature target proteins from living cells, candidate proteins are often analyzed with PMF and MS/MS ion search methods in combination with computational search routines based on bioinformatics. In contrast to shotgun proteomics, which is widely used to identify proteins, proteomics based on the analysis of N- and C-terminal amino acid sequences (terminal proteomics) should render higher fidelity results because of the high information content of terminal sequence and potentially high throughput of the method not requiring very high sequence coverage to be achieved by extensive sequencing. In line with this expectation, we review recent advances in methods for N- and C-terminal amino acid sequencing of proteins. This review focuses mainly on the methods of N- and C-terminal analyses based on MALDI-TOF MS for its easy accessibility, with several complementary approaches using LC/MS/MS. We also describe problems associated with MS and possible remedies, including chemical and enzymatic procedures to enhance the fidelity of these methods.

For selective C-terminal derivatization of peptides and proteins, we have devised a method for activating the C-terminal carboxyl group by extending the oxazolone chemistry. A mixture of formic acid and acetic anhydride was found to be effective for the formation of an oxazolone, which was converted to an active ester in situ in the presence of a phenol or an N-hydroxide. In particular, the resulting active ester with pentafluorophenol facilitated the subsequent reaction with an amine and the hydrazine derivative to yield the C-terminal amide and hydrazide, respectively. The peptides thus coupled with arginine methyl ester or 2-hydrazino-2-imidazoline containing the guanidino moiety exhibited the positive-ion peaks in matrix-assisted laser desorption/ionization (MALDI) mass spectra with appreciably enhanced intensities. As expected from the reaction mechanism, the carboxyl groups of aspartic and glutamic acid residues were not modified, while the amino groups that could react with the activated peptides were concomitantly protected by formylation. The MALDI peaks corresponding to the C-terminal peptide fragments of proteins were specifically enhanced, discriminating against those from internal peptides that were not tagged with a positive charge. In favorable cases, the C-terminal peptide fragments were clearly discerned by MALDI-MS after chymotryptic digestion and were identified by their MALDI postsource decay analysis. Based on these results, we suggest a method for C-terminal sequencing of a protein.

A high-throughput method for sequencing of N termini of proteins by using postsource decay (PSD) of matrix-assisted laser desorption/ionization mass spectrometry has been developed. After a protein blotted on the PVDF membrane was successively reduced, S-alkylated, and guanidinated, its N-amino group was coupled to biotinylcysteic acid. The protein was then extracted from the membrane and digested with trypsin. The derivatized N-terminal fragment was then specifically isolated from the tryptic digest with avidin resins, and its de novo sequencing was successfully performed by PSD utilizing a sulfonic acid group introduced to the N terminus.

The heat transport characteristics of liquid 3He and 3He 4He liquid mixtures (37% 3He, 15% 3He) were measured under saturated vapour pressure between 0.5 K and 0.7 K. The temperature difference ΔT between the heated copper surface and the liquid just above the surface was measured as a function of the heat flux q ˙ . The transition from the convection and conduction region to the nucleate boiling region was observed clearly for each sample. The curves of q ˙ versus ΔT for increasing and decreasing heat flux reveal no hysteresis effect at 0.5 K, but one at 0.7 K.

Recombinant holo Thermus thermophilus [7Fe-8S] ferredoxin was synthesized by cloning from Thermus thermophilus HB8 gene. A specific sequence (Pro-His-Val-Ile) at the N-terminus of the recombinant ferredoxin was determined by a rapid and highly sensitive mass spectral method using a novel Ru(II) Edman reagent, [(tpy)Ru(tpy-C6H4−NCS)](PF6)2 (tpy = terpyridine). The formation of the recombinant holoTtFd was established by the characteristic absorptions and CD extrema as [7Fe-8S] ferredoxin. The catalytic electron-transfer reactivity of the [7Fe-8S] ferredoxin between ferredoxin-NADP+ reductase and cytochrome c was recognized. Keywords: ESI−MS • protein identification • amino acid sequencing • Ru(II) Edman reagent • [7Fe-8S] ferredoxin

In relation to construction of the copper nuclear demagnetization stage, the residual resistance ratios (RRR) of ultra pure (7N) copper plates after annealing at various conditions were measured. Ultra pure copper is so active that even a small amount of oxygen diffused into the copper decreases the RRR value considerably. However, if oxidized layers were formed on the copper surface at an early stage of the annealing, the RRR value increased up to 10 4 after annealing.

Publisher Summary This chapter helps in constructing a simple dilution refrigerator without 1K pot, and it investigates its basic characteristics extensively. Several features peculiar to this type of the dilution machine have become clear for the first time and they are discussed in detail. The 3He-4He dilution refrigerator without 1K pot liquefies the circulating 3He by Joule-Thomson expansion and by making use of refrigeration capacity of the still and the 3He gas evaporated from it. The highest 3He inlet pressure is less than 1.4 atm, which enables one to circulate 3He without a compressor. The lowest temperature of the mixing chamber is 19.8 mK and highest temperature is 570 mK. The 3He circulation rate is limited in the range of 40 ∼ 70 μ mol/sec. Over this rate, the mixing chamber temperature becomes unstable.

Publisher Summary Among many modifications of the pulse tube refrigerator, the double-inlet type devised has great advantages in its improved performance and its simplicity. This chapter investigates the performance of one-stage double-inlet pulse tube refrigerator in detail. One-stage double-inlet pulse tube refrigerator generates a low temperature 23.9 K. Anomalous temperature variation occurred in the pulse tube against the bypass valve opening, which is considered to be common in this type of the pulse tube refrigerator. In order to make clear the reason for this anomaly, experiments are performed and reasonable conclusion is obtained. With opening the bypass valve, the temperature distribution along the pulse tube changes from the one with low temperature (∼220 K ) at the position T4 of the pulse tube to the other with high temperature ( ∼270 K ) at the position T4.

Publisher Summary This chapter illustrates that a great deal of experimental and theoretical studies on superfluid turbulence have been performed. However, the chapter investigates the influence of the vortex line on pressure dissipation This chapter measures the temperature difference and the pressure difference through He II in a capillary glass tube. It calculates the quantized vortex line density from the temperature difference data using the numerical scaling coefficients. The pressure difference data give the information of interaction between the vortex line and the tube wall. The pressure dissipation caused by the vortex line is only one tenth of mutual friction between the vortex line and the normalfluid. In the terms of eddy viscosity for superfluid velocity field, it calculates the coefficient of the interaction between the vortex line and the superfluid.

A novel N-terminal-labeling reagent [(tpy)Ru I I -(tpyCONSu)] 2 + (= -COONSu) provides an efficient and effective N-terminal sequencing of peptides and proteins in MALDI-TOF-MS/MS (PSD) analysis, which shows a n fragments predominantly without any C-terminal-fragment ions.

Superconducting phase transitions of quasi-one-dimensional compound have been studied by the measurement of AC magnetic susceptibility χ ′ . The χ ′ of powdered sample decreased stepwise, which was characterized by two transition temperatures T C1 and T C2 . The T C1 is considered to be the transition within the grains, and T C2 is the inter-grain transition. Under the magnetic field, T C2 shifted to the lower temperature. The magnetic field dependence of T C2 is presented.