Basic studies toward ultrafast soft x-ray photoelectron diffraction; its application to probing local structure in iodobenzene molecules.

Ultrafast x-ray photoelectron diffraction (UXPD) for free molecules has a promising potential to probe the local structures of the molecules in an element-specific fashion. Our UXPD scheme consists of three steps: (1) near-infrared laser (NIR) with ns pulse duration aligns sample molecules, (2) ultra-violet laser with fs pulse duration pumps the aligned molecules, and (3) soft x-ray free-electron laser (SXFEL) with fs pulse duration probes the molecules by measuring x-ray photoelectron diffraction (XPD) profiles. Employing steps of (1) and (3), we have measured I 3d XPD profiles from ground state iodobenzene aligned by the NIR laser with the SXFEL. Then, we have intensively calculated I 3d XPD profiles with density functional theory, taking degrees of alignments of the molecules into account, to extract a distance between C and I atoms in iodobenzene from the experimental I 3d XPD profiles. Although we have failed to determine the distance from the comparison between the experimental and theoretical results, we have succeeded in concluding that the degeneracies of the initial state eliminate the sensitivity on molecular structure in the I 3d XPD profiles. Thus, the observation of fine structures in the XPD profiles could be expected, if a nondegenerate molecular orbital is selected for a probe of UXPD. Finally, we have summarized our criteria to perform UXPD successfully: (1) to use SXFEL, (2) to prepare sample molecules with the degree of alignment higher than 0.8, and (3) to select a photoemission process from a nondegenerate inner-shell orbital of sample molecules.

Fast-ion-induced secondary ion emission from submicron droplet surfaces studied using a new coincidence technique with forward-scattered projectiles.

A mass spectrometric study of secondary ions emitted from droplet surfaces by MeV-energy heavy ion impact was performed to investigate fast-ion-induced molecular reaction processes on liquid surfaces. Herein, a new coincidence technique was developed between secondary ions and scattered projectile ions at a small forward angle. The advantages of this technique were demonstrated by measurement of the collision between 4-MeV C3+ and ethanol droplets. Secondary ion emission probabilities were obtained directly from the coincidence data. Notably, this technique enabled positive fragment ions that had not been identified in previous measurements to be observed by suppressing the strong background originating from gas-phase molecules more than 104-fold. H+, H3O+, C2H5 +, and C2H5O+ were found to be produced as major positive fragment ions, in addition to minor fragments H2 +, C2H3 +, and CH2OH+. Production of these ions suggests that competition between rapid hydrogen ion emission from multiply ionized states and intermolecular proton transfer accompanied by fragmentation through protonated ethanol occurs after fast heavy-ion collisions. Clarification of the positive fragment ions also revealed the characteristic features of negative ions. Negative ions were realized to exhibit higher degrees of fragmentation and reactivity compared with positive ions. Furthermore, the energy loss by forward-scattered ions during droplet penetration was used to evaluate the target thickness at a submicron level. Variations in secondary ion yield, mass distribution, and kinetic energies depending on the penetration length were observed below 1 µm. These results highlight the unknown mechanism of these "submicron effects" observed in secondary ion emission processes as a new phenomenon.

A cryogenic linear ion trap beamline for providing keV ion bunches.

A new cryogenic linear ion trap beamline has been constructed and commissioned, which serves to inject cold molecular and cluster ions into the RIKEN cryogenic electrostatic ring (RICE). Ions are created with an electrospray ion source, and a quadrupole mass filter is used for mass-selection prior to trap injection. The radio frequency octupole ion trap can be continuously loaded with ions and features a fast ion extraction mode to create short ion bunches with tens of µs duration. We report here on the simulations and development of the ion trap beamline and validate performance with the moderately heavy molecular cation methylene blue. Characterization of the novel trap design with additional wedge-shaped electrodes was carried out, which includes the determination of the temporal and spatial shape of the ion bunch and the total number of ions after extraction. Finally, these ion bunches are synchronized with the switching of a pulsed high-voltage acceleration device downstream of the trap, where the ions obtain a kinetic energy of up to 20 keV. The preparation and control of the keV ion beam are demonstrated for the ion injection into RICE.

Effects of herpes simplex virus vectors encoding poreless TRPV1 or protein phosphatase 1α in a rat cystitis model induced by hydrogen peroxide.

Enhanced afferent excitability is considered to be an important pathophysiological basis of interstitial cystitis/bladder pain syndrome (IC/BPS). In addition, transient receptor potential vanilloid-1 (TRPV1) receptors are known to be involved in afferent sensitization. Animals with hydrogen peroxide (HP)-induced cystitis have been used as a model exhibiting pathologic characteristics of chronic inflammatory condition of the bladder. This study investigated the effect of gene therapy with replication-defective herpes simplex virus (HSV) vectors encoding poreless TRPV1 (PL) or protein phosphatase 1 α (PP1α), a negative regulator of TRPV1, using a HP-induced rat model of cystitis. HSV vectors encoding green fluorescent protein, PL or PP1α were inoculated into the bladder wall of female rats. After 1 week, 1% HP or normal saline was administered into the bladder, and the evaluations were performed 2 weeks after viral inoculation. In HP-induced cystitis rats, gene delivery of PL or PP1α decreased pain behavior as well as a reduction in the intercontraction interval. Also, both treatments reduced nerve growth factor expression in the bladder mucosa, reduced bladder inflammation characterized by infiltration of inflammatory cells and increased bladder weight. Taken together, HSV-mediated gene therapy targeting TRPV1 receptors could be effective for the treatment of IC/BPS.

Cooling dynamics of photoexcited C6(-) and C6H(-).

We report conclusive evidence of an efficient cooling mechanism via the electronic radiative transitions of hot small molecular anions isolated in vacuum. We stored C6(-) and C6H(-) in an ion storage ring and observed laser-induced electron detachment with delays up to several milliseconds. The terminal hydrogen atom caused a drastic change in the decay profiles. The decay of photoexcited C6H(-) is slow and nonexponential, which can be explained by depletion cooling, whereas that for C6(-) occurs extremely fast, on a time scale below 0.1 ms and can only be explained by electronic radiative cooling via low-lying electronic excited states.

(99m)Tc-mercaptoacetyl triglycine renography to monitor renal transplant function among kidneys from donors after cardiac death.

PURPOSE: Delayed graft function usually occurs after kidney transplantation from donors after cardiac death, It is important to monitor graft function during the anuric period, but there have been few useful tools. Consequently, we evaluated the availability of (99m)-Tc mercaptoacetyltriglycine (MAG3) renography. METHODS: Thirty-two patients underwent renal transplantation from donors after cardiac death between June 2, 2005, and April 14, 2011. One patient was excluded due to an acute rejection episode which developed during the dialysis period. The first (99m)Tc-MAG3 renogram was performed as early as possible after the operation and repeated until the patient was weaned from dialysis. The corrected tubular extraction rate (cTER; mL/min/1.73 m(2)) was calculated; it represents the MAG3 clearance corrected by body surface area. RESULTS: cTER was low immediately after transplantation, but increased gradually until the patient was weaned from dialysis. A significant correlation was observed between early cTER and the period of dialysis-dependence (r = -0.677, P < .001) as well as the short-term best corrected creatinine clearance (r = 0.526, P = .002). CONCLUSION: We observed that graft function can be monitored by routinely performing (99m)Tc-MAG3 renography after transplantation of kidneys from donors after cardiac death.

Atelocollagen-associated autologous chondrocyte implantation for the repair of chondral defects of the knee: a prospective multicenter clinical trial in Japan.

BACKGROUND: New tissue-engineering technology was developed to create a cartilage-like tissue in a three-dimensional culture using atelocollagen gel. The minimum 2-year followup outcome of transplanting autologous chondrocytes cultured in atelocollagen gel for the treatment of full-thickness defects of cartilage in knees was reported from the single institution. The present multicenter study was conducted to determine clinical and arthroscopic outcomes in patients who underwent atelocollagen-associated autologous chondrocyte implantation for the repair of chondral defects of the knees. METHODS: At six medical institutes in Japan, we prospectively evaluated the clinical and arthroscopic outcomes of transplanting autologous chondrocytes cultured in atelocollagen gel for the treatment of full-thickness defects of cartilage in 27 patients (27 knees) with cartilage lesions on a femoral condyle or on a patellar facet over 24 months. RESULTS: The Lysholm score significantly increased from 60.0 +/- 13.7 points to 89.8 +/- 9.5 points (P = 0.001). Concerning the ICRS grade for arthroscopic appearance, 6 knees (24%) were assessed as grade I (normal) and 17 knees (68%) as grade II (nearly normal). There were few adverse features, except for detachment of the graft in two cases. CONCLUSIONS: We concluded that transplanting chondrocytes in a newly formed matrix of atelocollagen gel can promote restoration of the articular cartilage of the knee.

Alteration of N-glycans related to articular cartilage deterioration after anterior cruciate ligament transection in rabbits.

OBJECTIVE: Osteoarthritis (OA) is the most common of all joint diseases, but the molecular basis of its onset and progression is controversial. Several studies have shown that modifications of N-glycans contribute to pathogenesis. However, little attention has been paid to N-glycan modifications seen in articular cartilage. The goal of this study was to identify disease specific N-glycan expression profiles in degenerated cartilage in a rabbit OA model induced by anterior cruciate ligament transection (ACLT). METHODS: Cartilage samples were harvested at 7, 10, 14, and 28 days after ACLT and assessed for cartilage degeneration and alteration in N-glycans. N-Glycans from cartilage were analyzed by high performance liquid chromatography and mass spectrometry. RESULTS: Histological analysis showed that osteoarthritic changes in cartilage occurred 10 days after ACLT. Apparent alterations in the N-glycan peak pattern in cartilage samples were observed 7 days after ACLT, and overall N-glycan changes in OA reflected alterations in both sialylation and fucosylation. These changes apparently preceded histological changes in cartilage. CONCLUSION: These results indicate that changes in the expression of N-glycans are correlated with OA in an animal model. Understanding mechanisms underlying changes in N-glycans seen in OA may be of therapeutic value in treating cartilage deterioration.

Chitosan-based hyaluronan hybrid polymer fibre scaffold for ligament and tendon tissue engineering.

To establish medical use of tissue engineering technology for ligament and tendon injuries, a scaffold was developed which has sufficient ability for cell growth, cell differentiation, and mechanical properties. The scaffold made from chitosan and 0.1 per cent hyaluronic acid has adequate biodegradability and biocompatibility. An animal experiment showed that the scaffold has less toxicity and less inflammation induction. Furthermore, in-vivo animal experiments showed that the mechanical properties of the engineered ligament or tendon had the possibility to stabilize the joint. It was shown that newly developed hybrid-polymer fibre scaffold has feasibility for joint tissue engineering.

Inverted V-shaped high tibial osteotomy compared with closing-wedge high tibial osteotomy for osteoarthritis of the knee. Ten-year follow-up result.

We compared the results ten years after an inverted V-shaped high tibial osteotomy with those of a historical series of conventional closing-wedge osteotomies. The closing-wedge series consisted of 56 knees in 51 patients with a mean follow-up of 11 years (10 to 15). The inverted V-shaped osteotomy was evaluated in 48 knees in 43 patients at a mean follow-up of 14 years (10 to 19). All the patients were scored using the Japanese Orthopaedic Association rating scale for osteoarthritis of the knee and radiological assessment. The pre-operative grade of osteoarthritis was similar in both groups. Post-operatively, the knee function score was graded as satisfactory in 63% (35) of the closing-wedge group but in 89% (43) of the inverted V-shaped osteotomy group. Post-operative radiological examination showed that delayed union and loss of correction occurred more often after a closing-wedge osteotomy than after an inverted V-shaped procedure. Our study suggests that the inverted V-shaped osteotomy may offer more dependable long-term results than traditional closing-wedge osteotomy.

Extracellular matrix modulates expression of cell-surface proteoglycan genes in fibroblasts.

Cell-surface proteoglycans are involved in many functions, including interactions with components of the extracellular microenvironment. They also act as coreceptors that bind and modify the actions of various growth factors, cytokines, and the extracellular matrix (ECM). This study investigated the regulation by the ECM of the expression of cell-surface proteoglycans (CD44, syndecan-1-4, betaglycan, glypican-1). We examined the changes in the expression levels of cell-surface proteoglycan genes in intact tendon, monolayer culture, and under various culture conditions. There was a significant increase in the expression of CD44 and syndecan-4 mRNAs during cell isolation from the tendon. With the switch to a 3D culture environment, there was a significant increase in the expression of CD44 at each passage point relative to its expression in 2D at those passage points. Syndecan-4 mRNA also increased steadily at each passage point in 3D culture environment. This influence on cell surface proteoglycans gene expression may indicate that collagen gel culture mimics in vivo tendon environment. This study provides further insight into the regulation of cell-surface proteoglycans in ligament and tendon fibroblasts by the ECM and 3D culture conditions.

Negative correlation between bone mineral density and TSH receptor antibodies in male patients with untreated Graves' disease.

INTRODUCTION: Although it has been established that hyperthyroidism leads to reduced bone mineral density (BMD), with accelerated bone turnover promoting bone resorption in female patients, there is a dearth of data for male patients with hyperthyroidism. This study evaluated BMD and bone metabolism in male patients with Graves' disease. METHODS: The study included 56 Japanese male patients with newly diagnosed Graves' disease and 34 normal Japanese male control subjects of similar age and body mass index. We used dual energy x-ray absorptiometry to measure BMD at sites with different cortical/cancellous bone ratios (lumbar spine, femoral neck, and distal radius). RESULTS: At the lumbar spine and the distal radius, BMD and T-scores were significantly lower for patients than for controls. At the femoral neck, on the other hand, the same values were relatively, but not significantly, lower in patients than in controls. However, Z-scores at all three sites were significantly lower for patients than for controls. The Z -score at the distal radius of patients was significantly lower than that at their lumbar spine and femoral neck. In addition, Z-score at the distal radius correlated negatively with age, free thyroxine, thyroid stimulating hormone receptor antibodies, thyroid stimulating antibody, and urinary N-terminal telopeptide of type I collagen normalized by creatinine. CONCLUSIONS: These results indicate a high prevalence of cortical bone loss in male patients with Graves' disease, especially elderly patients. We conclude that BMD measurement is crucial in all Graves' disease patients regardless of their gender and that the radial BMD as well as BMD at the lumbar spine and femoral neck should be monitored to effectively prevent bone loss and subsequent fracture.

Decreased bone mineral density at the distal radius, but not at the lumbar spine or the femoral neck, in Japanese type 2 diabetic patients.

The purpose of this study is to assess the association between type 2 diabetes and bone mineral density. This study included 145 Japanese patients (64 men and 81 women) with type 2 diabetes and 95 non-diabetic control subjects (41 men and 54 women) of similar age. We measured bone mineral density (BMD) at the sites with different cortical/cancellous bone ratio (lumbar spine, femoral neck, and distal radius) using dual-energy X-ray absorptiometry. BMD and Z score at the distal radius were significantly lower in type 2 diabetic patients than those in control subjects, and in type 2 diabetic patients, the Z score at the distal radius was lower than that at their own lumbar spine and femoral neck. In type 2 diabetic patients, negative correlation between BMD and the mean HbA1c during the previous 2 years was found significantly at the distal radius in both genders and at the femoral neck in women. These results indicate the selective cortical bone loss in type 2 diabetes and suggest the importance of also determining BMD at the radius and keeping good metabolic control to prevent bone loss in type 2 diabetic patients.

Chemiluminescent visualization of superoxide generated by Candida albicans.

The high toxicity of reactive oxygen species (ROS) suggested a possible role in the pathogenicity of human pathogenic fungi. We previously reported a chemiluminescence method for measuring ROS generation in Candida albicans. In the present study, we attempted to visualize the ROS, superoxide anion radical (O2-), generated by paraquat (PQ)-stimulated C. albicans using methyl-Cypridina-luciferin analog (MCLA) as a chemiluminescence probe. Colonies of a wild-type C. albicans parent strain and its respiration-deficient mutant grown on agar plates were overlaid with a mixture of PQ and MCLA solutions. MCLA-dependent light emission from the colonies was recorded with a Hamamatsu ultralow-light-imaging apparatus with a CCD camera in a light-tight box. In the wild-type strain, marginal regions of growing colonies were strongly illuminated. The light emission from the colonies was extinguished by superoxide dismutase (SOD), proving that the light emission was strictly due to the superoxide anion. However, colonies of the respiration-deficient mutant poorly generated superoxide. Chemiluminecence measurements by a luminometer showed vigorous superoxide generation by the exponential phase cells of the parent strain but weak generation by the stationary phase cells. In the mutant, superoxide generation was weak compared with the parent strain. These results indicate that expansion of the colonies was due to the actively respiring cells located in the marginal regions. To our knowledge, the present report is the first chemiluminescent visualization of ROS including superoxide generated by C. albicans.

[Acute exacerbations due to Streptococcus pneumoniae in chronic lower respiratory tract infections during long-term macrolide therapy].

In Japan, long-term 14-membered macrolide administration is chosen as a first line therapy against chronic lower respiratory tract infections (CLRTIs) such as diffuse panbronchiolitis, bronchiectasis and chronic bronchitis. However, sometimes acute exacerbations occur in these cases, even if therapy is effective. We investigated 18 episodes of CLRTIs exacerbations that were caused by Streptococcus pneumoniae during long-term macrolides therapy from 1991 to 1999 to clarify the clinical features and prevalence of antimicrobial resistance in S. pneumoniae. Exacerbations did not occur only in winter season, but also in other seasons. Among 18 episodes of exacerbation, only 7 episodes (39%) revealed infiltration in chest roentogenogram and few episodes revealed marked elevations of inflammation markers in laboratory data. Intermediate resistance or resistance rates of S. pneumoniae isolated from sputum or transtracheal aspiration were 100% to erythromycin, 67% to clindamycin or minocycline, 11% to ampicillin, and 0% to cephazoline or imipenem. Coresistance to erythromycin, clindamycin and minocycline was seen in a half of the episodes. Resistance was not correlated with the duration of macrolides administration. All episodes were mainly treated with beta-lactam agents or fluoroquinolones and cured successfully. These findings suggest that acute exacerbations in CLRTIs caused by S. pneumoniae during long-term macrolides therapy do not reveal severe clinical aspects and can be treated successfully at present, but attention should be paid to the trend of antibiotic susceptibility in S. pneumoniae.

[Clinical effectiveness of intra-arterial chemotherapy combined with degradable starch microspheres (DSM) for liver metastases and prediction of effectiveness by diagnostic imaging].

The aim of this study was to evaluate the clinical effectiveness of intra-arterial chemotherapy combined with degradable starch microspheres (DSM) for liver metastases and the possibility of predicting the effectiveness of the chemotherapy by pretreatment diagnostic imaging. The subjects were 67 patients with metastatic liver cancer, treated with Seldinger method via the left brachial artery, and tumor selective hepatic injection using a micro-catheter. The early response rate was 38.7% for colorectal cancer, 42.8% for gastric cancer, 16.7% for bile tract cancer and 80% for uterine cancer. The relationship between effectiveness and the tumor occupation rate in the liver estimated from pretreatment CT images was not significant, but the degree of tumor stain in the early phase of contrast enhancement CT correlated well with early responsiveness of the liver metastases for this treatment. This suggests the possibility of pretreatment prediction of the effectiveness of intra-arterial chemotherapy combined with DSM for metastatic liver tumors.

Interleukin-10 and interferon-gamma levels within the peritoneal cavity of patients with gastric cancer.

BACKGROUND AND OBJECTIVES: Immune status in the peritoneal cavity of patients with gastric cancer remains largely unknown. To clarify the clinical significance of the host immune response within the peritoneal cavity, we examined the levels of interferon-gamma (IFN-gamma), a type 1 cytokine, and interleukin-10 (IL-10), a type 2 cytokine, in peritoneal washings obtained from patients with gastric cancer. METHODS: Both the concentrations of IFN-gamma and of IL-10 in peritoneal washings obtained during surgery from 56 patients with gastric cancer were determined by an enzyme-linked immunosorbent assay. RESULTS: The IFN-gamma level was not correlated with the IL-10 level. The IL-10 level increased in a stage-dependent manner. The high IL-10 level correlated with an unfavorable outcome, whereas there was no relationship between the IFN-gamma level and survival rate. However, among the stage III-IV cancer patients, the high IFN-gamma level correlated with a favorable outcome, while there was no relationship between the IL-10 level and survival rate. CONCLUSION: Although the IL-10 level increases with tumor progression, the outcome of patients with advanced gastric cancer may be affected by the IFN-gamma level, but not by the IL-10 level, in the peritoneal cavity.

Preoperative serum interleukin-18 level as a postoperative prognostic marker in patients with gastric carcinoma.

BACKGROUND: Interleukin-18 (IL-18), a recently described cytokine produced mainly by macrophages, stimulates interferon-gamma (IFN-gamma) production by natural killer cells and T cells. Although it has been reported that serum IL-18 levels are higher in patients with advanced tuberculosis and acute graft-versus-host disease compared with normal controls, the authors found no reports regarding serum IL-18 levels in patients with malignant solid tumors. The purpose of this study was to determine serum IL-18 levels and their clinical significance in patients with gastric carcinoma. METHODS: Peripheral blood samples were obtained from 94 patients with gastric carcinoma who underwent curative surgery and from 50 healthy volunteers. The serum IL-18 level, the IFN-gamma, level, and the Helicobacter pylori (HP) serology status were determined in each sample with an enzyme-linked immunosorbent assay. RESULTS: The mean serum IL-18 level for all patients was significantly higher compared with the mean level in healthy volunteers (P < 0.01). IFN-gamma titers were below the level of detection in all samples tested. When the patients were subdivided into groups, it was found that the serum IL-18 level in patients with Stage II and III disease was significantly higher compared with the level found in healthy volunteers (P < 0.01). The serum IL-18 level decreased after patients underwent surgical resection. However, there was no significant difference in the serum IL-18 level between healthy controls and patients with Stage I or IV disease. Patients with IL-18 levels >or= 310 pg/mL (i.e., equal to or greater than the mean levels +/- 1 standard deviation in the healthy volunteers) experienced a significantly lower survival rate compared with patients who had IL-18 levels < 310 pg/mL after undergoing surgery (P < 0.05) despite a lack of any discernible difference in clinicopathologic factors between the two groups. The serum IL-18 level was identified as an independent postoperative prognostic factor in multivariate survival analysis using a Cox proportional hazards model (hazard ratio, 4.89; P = 0.01). There was no significant correlation between HP serology status and serum IL-18 levels. CONCLUSIONS: The preoperative serum IL-18 level may represent a significant postoperative prognostic determinant in patients with gastric carcinoma. Its function in the host immune system remains to be elucidated.