The dynamics of NO rebinding in hemoglobin (Hb) was directly observed using femtosecond mid-IR spectroscopy after photodeligation of NO from HbNO in D2O at 283 K. Time-resolved spectra of bound NO appeared to have a single feature peaked at 1616 cm-1 but were much better described by two Gaussians with equal intensities but different rebinding kinetics, where the feature at 1617 cm-1 rebinds faster than the one at 1614 cm-1. It is possible that the two bands each correspond to one of two subunit constituents of the tetrameric Hb. Transient absorption spectra of photodeligated NO revealed three evolving bands near 1858 cm-1 and their red-shifted replicas. The red-shifted replicas arise from photodeligated NO in the vibrationally excited v = 1 state. More than 10% of the NO was dissociated into the vibrationally excited v = 1 state when photolyzed by a 580-nm pulse. The three absorption bands for the deligated NO could be attributed to three NO sites in or near the heme pocket. The kinetics of the three transient bands for the deligated NO, as well as the recovery of the bound NO population, was most consistent with a kinetics scheme that incorporates time-dependent rebinding from one site that rapidly equilibrates with the other two sites. The time dependence results from a time-dependent rebinding barrier due to conformational relaxation of protein after deligation. By assigning each absorption band to a site in the heme pocket of Hb, a pathway for rebinding of NO to Hb was proposed.

Diabetic kidney disease (DKD) is the leading cause of end-stage renal disease worldwide. Excess accumulation of extracellular matrix and the epithelial-to-mesenchymal transition contribute to renal fibrosis, which is associated with DKD. The present study examined whether delayed treatment with human umbilical cord blood-derived stem cells (hUCB-SC) showed a therapeutic effect on DKD progression. Experimental diabetes was induced by intraperitoneal injection of streptozotocin (STZ; 50 mg/kg) into 6-week-old male Sprague-Dawley rats. Age-matched control rats received an equivalent volume of sodium citrate buffer alone. At 4 weeks after the STZ injection when diabetic renal injury had developed, hUCB-SC were administered (1 × 10(6) cells/rat) through the tail vein. Four weeks after administering the hUCB-SC, rats were sacrificed and we measured indices of DKD, including urinary protein excretion as well as fibronectin, α-smooth muscle actin (α-SMA), and E-cadherin mRNA, and protein expression. Diabetic rats developed significantly increased urinary protein excretion and renal hypertrophy compared to those in control rats. Renal expression of fibronectin and α-SMA mRNA, and protein were increased significantly in diabetic rats compared to those in the controls. E-cadherin protein expression in diabetic kidneys decreased significantly. Intravenously administered hUCB-SC effectively reduced proteinuria, renal fibronectin, and α-SMA up-regulation, as well as renal E-cadherin down-regulation in diabetic rats without a significant effect on blood glucose. Engrafted hUCB-SC in diabetic kidneys were confirmed by human DNA PKcs. The results demonstrated that delayed treatment with hUCB-SC attenuated the progression of diabetic renal injury.

Fractalkine (CX3CL1) is a unique chemokine that functions not only as a chemokine but also as an adhesion molecule. Fractalkine plays an important role in the recruitment of macrophages into the kidneys by binding to its specific receptor CX3CR1, and renal fractalkine expression was shown to be increased in chronic renal allograft rejection. Considering that microcapillary inflammation is a key feature of chronic renal allograft rejection, the present study examined whether monocytes bind to mesangial cells cultured in the presence of lipopolysaccharide (LPS) through fractalkine/CX3CR1 in order to understand their regulation with respect to inflammation-induced renal allograft dysfunction. Mouse mesangial cells were stimulated with LPS in the presence or absence of fractalkine or CX3CR1 siRNA. Calcein-AM-labeled monocytes were used to evaluate monocyte binding. Fractalkine and CX3CR1 mRNA and protein expression were measured by real-time quantitative polymerase chain reaction and enzyme-linked immunosorbent assay, respectively. LPS at 100 ng/mL significantly increased monocyte binding to mesangial cells. Each siRNA against fractalkine or CX3CR1 effectively inhibited LPS-induced monocyte-mesangial cell binding. Fractalkine and CX3CR1 mRNA expression were enhanced in mesangial cells stimulated with LPS. Fractalkine protein synthesis in media and lysate of mesangial cells were also induced by LPS. These results demonstrated that LPS induces monocyte-mesangial cell binding through the fractalkine/CX3CR1 system and suggested that fractalkine/CX3CR1 system may contribute to renal inflammation leading to chronic renal allograft rejection.

Mesangial cell proliferation is one of the main features of chronic renal allograft rejection. One unique feature of fractalkine (CX3CL1) is its existence as both a membrane-tethered and a soluble form. Fractalkine expression is increased in acute and chronic allograft rejection. However, its role in mesangial cell proliferation has not yet been clearly explored. Thus, the present study examined whether fractalkine induced mesangial cell proliferation through production of reactive oxygen species (ROS) and activation of mitogen-activated protein kinase (MAPK), two known mediators of mesangial cell proliferation. Growth-arrested and synchronized mouse mesangial cells were stimulated with fractalkine in the presence versus absence of inhibitors against ROS, extracellular signal-regulated protein kinase (ERK), and p38 MAPK. Cell proliferation was assessed by methylthiazoletetrazolium assay, dichlorofluorescein (DCF)-sensitive cellular ROS production by a fluorometer, and MAPK activation by Western blot analysis. Fractalkine (10-50 ng/mL) significantly increased mesangial cell proliferation at 24 hours in a dose-dependent manner, an effect that was abrogated by the ROS and MAPK inhibitors. Fractalkine (50 ng/mL) also induced cellular ROS production and activation of ERK1/2 and p38 MAPK in mesangial cells. These results demonstrated that fractalkine can induce mesangial cell proliferation through production of cellular ROS and activation of MAPK.

Diabetes, whether it occurs before or after transplantation, plays an important role to decrease graft function and survival. In addition renal lipid accumulation has been suggested to play a role in the development and progression of chronic renal allograft rejection. Intracellular lipid accumulation is governed by a balance between the influx and efflux of lipid. Cholesterol transporters, such as scavenger receptor (SR)-A1, CD36, and ATP binding cassette (ABC) A1 and G1 (ABCG1), coordinate to regulate cellular lipid status. Therefore, in the present study, we examined whether high glucose caused lipid accumulation in mesangial cells as a result of altered cholesterol transporters. Mouse mesangial cells were stimulated with 30 mmol/L D-glucose (high glucose); 100 μmol/L oleic acid (OA) used as a positive control. Cellular lipid accumulation was measured by Oil Red O staining. Protein and mRNA expression of cholesterol influx (SR-A1 and CD36) and efflux (ABCA1 and ABCG1) transporters were evaluated using Western blot analysis and real-time quantitative polymerase chain reaction, respectively. High glucose was shown to significantly increase lipid accumulation in mesangial cells at 24 hours as was observed for OA. SR-A1 and CD36 mRNA expression levels were 1.5-fold and 3.5-fold higher, respectively, in high glucose-stimulated than control mesangial cell, whereas ABCG1 mRNA expression decreased to 60% of controls; however, there was no decrease in ABCA1 mRNA. Altered protein expression of each transporter in mesangial cells cultured under conditions of high glucose concentrations was consistent with mRNA expression. Osmotic control using mannitol did not significantly affect any of the measured parameters in the present study. These results demonstrated that high glucose, in itself, can induce mesangial lipid accumulation; this effect may be associated with an impaired balance between the influx and efflux of cholesterol.

In this study, we tested several hypotheses related to changes in finger interaction and multi-finger synergies during multi-finger force production tasks in Parkinson's disease. Ten patients with Parkinson's disease, mostly early-stage, and eleven healthy control subjects participated in the study. Synergies were defined as co-varied adjustment of commands to fingers that stabilized the total force produced by the hand. Both Parkinson's disease patients and control subjects performed accurate isometric force production tasks with the fingers of both the dominant and non-dominant hand. The Parkinson's disease patients showed significantly lower maximal finger forces and higher unintended force production (enslaving). These observations suggest that changes in supraspinal control have a major effect on finger individuation. The synergy indices in the patients were weaker in both steady-state and cyclic force production tasks as compared to the controls. These indices also were stronger in the left (non-dominant) hand in support of the dynamic-dominance hypothesis. Half of the patients could not perform the cyclic task at the highest frequency (2 Hz). Anticipatory adjustments of synergies prior to a quick force pulse production were delayed and reduced in the patients as compared to the controls. Similar differences were observed between the asymptomatic hands of the patients with symptoms limited to one side of the body and matched hands of control subjects. Our study demonstrates that the elusive changes in motor coordination in Parkinson's disease can be quantified objectively, even in patients at a relatively early stage of the disease. The results suggest an important role of the basal ganglia in synergy formation and demonstrate a previously unknown component of impaired feed-forward control in Parkinson's disease reflected in the reduced and delayed anticipatory synergy adjustments.

Use of bacteriophages as biocontrol agents is a promising tool for controlling pathogenic bacteria including antibiotic-resistant bacteria. Not only bacteriophages but also endolysins, the peptidoglycan hydrolyzing enzymes encoded by bacteriophages, have high potential for applications as biocontrol agents against foodborne pathogens. In this study, a putative endolysin gene was identified in the genome of the bacteriophage BPS13, which infects Bacillus cereus. In silico analysis of this endolysin, designated LysBPS13 showed that it consists of an N-terminal catalytic domain (PRGP domain) and a C-terminal cell wall binding domain (SH3_5 domain). Further characterization of the purified LysBPS13 revealed that this endolysin is an N-acetylmuramyl-L-alanine amidase, the activity of which was not influenced by addition of EDTA. In addition, LysBPS13 demonstrated remarkable thermostability in the presence of glycerol, and it retained its lytic activity even after incubation at 100 °C for 30 min. Taken together, these results indicate that LysBPS13 can be considered a favorable candidate for a new antimicrobial agent to control B. cereus. © 2012 Federation of European Microbiological Societies. Published by Blackwell Publishing Ltd. All rights reserved.

To achieve high performance in brain-computer interfaces (BCIs) using P300, most of the work has been focused on feature extraction and classification algorithms. Although significant progresses have been made in such signal processing methods in the lower layer, the issues in the higher layer, specifically determining the stimulus schedule in order to identify the target reliably and efficiently, remain relatively unexplored. In this article, we propose a systematic approach to compute an optimal stimulus schedule in P300 BCIs. Our approach adopts the partially observable Markov decision process (POMDP), which is a model for planning in partially observable stochastic environments. We show that the thus obtained stimulus schedule achieves a significant performance improvement in terms of the success rate, bit rate, and practical bit rate through human subject experiments.

OBJECTIVE: To evaluate the incidence, clinical characteristics and predicting factors for the development of paradoxical response in human immunodeficiency virus negative patients with isolated pleural tuberculosis (TB).DESIGN: A multicentre, retrospective cohort study including 458 patients who were diagnosed and treated with isolated pleural TB between March 2005 and February 2010.RESULTS: Paradoxical response developed in 72 patients (16%) with isolated pleural TB. The mean time to development of paradoxical response was 8.8 ± 6.4 weeks after initiation of anti-tuberculosis treatment. The main presentation of paradoxical response was aggravation of pre-existing pleural effusion in 58 patients (81%). However, the majority of the patients who developed paradoxical response had no associated symptoms (n = 49, 68%). In multiple logistic regression analysis, development of paradoxical response was independently associated with the proportion of eosinophils (adjusted OR 1.293, 95%CI 1.077-1.553) and protein concentrations (adjusted OR 0.590, 95%CI 0.397-0.878) in the pleural fluid at the time of diagnosis.CONCLUSION: Paradoxical response developed in 16% of the patients approximately 2 months after initiation of anti-tuberculosis treatment, presenting with aggravation of pre-existing pleural effusion. Development of paradoxical response was associated with the proportion of eosinophils and protein concentrations in the pleural fluid at the time of diagnosis.

Microbial fuel cell (MFC) technology offers a sustainable approach to harvest electricity from biodegradable materials. Energy production from MFCs have been demonstrated using external resistors or charge pumps, but such methods can only dissipate energy through heat or receive electrons passively from the MFC without any controllability. This study developed a new approach and system that can actively extract energy from MFC reactors at any operating point without using any resistors, especially at the peak power point to maximize energy production. Results show that power harvesting from a recirculating-flow MFC can be well maintained by the maximum power point circuit (MPPC) at its peak power point, while a charge pump was not able to change operating point due to current limitation. Within 18-hour test, the energy gained from the MPPC was 76.8 J, 76 times higher than the charge pump (1.0 J) that was commonly used in MFC studies. Both conditions resulted in similar organic removal, but the Coulombic efficiency obtained from the MPPC was 21 times higher than that of the charge pump. Different numbers of capacitors could be used in the MPPC for various energy storage requirements and power supply, and the energy conversion efficiency of the MPPC was further characterized to identify key factors for system improvement. This active energy harvesting approach provides a new perspective for energy harvesting that can maximize MFC energy generation and system controllability.