Robustness of large-area suspended graphene under interaction with intense laser.

Graphene is known as an atomically thin, transparent, highly electrically and thermally conductive, light-weight, and the strongest 2D material. We investigate disruptive application of graphene as a target of laser-driven ion acceleration. We develop large-area suspended graphene (LSG) and by transferring graphene layer by layer we control the thickness with precision down to a single atomic layer. Direct irradiations of the LSG targets generate MeV protons and carbons from sub-relativistic to relativistic laser intensities from low contrast to high contrast conditions without plasma mirror, evidently showing the durability of graphene.

Molecular dynamics simulation of ß-adrenoceptors and their coupled G proteins.

OBJECTIVE: G protein-coupled receptors (GPCRs) constitute the largest membrane proteins superfamily. However, the interactions between them and the coupled heterotrimeric G proteins were little known. To get a deeper view of how the receptor bound to the G protein, we carried out the molecular dynamics' simulations of human Beta2 adrenoceptors (ß1 and ß2) and G protein (s and I) alpha subunit complexes by homology modeling. MATERIALS AND METHODS: For homology modeling, the program modeller 9.11 was used with automodel module. Before dynamics simulation, the homology models were prepared by Protein Preparation Wizard module in Maestro 9.3. The Desmond program was used to perform molecular minimization and molecular dynamics simulation under OPLS-All atom 2005 force field with default parameters. RESULTS: The results offered us the mechanism vividly in molecular level: (1) GPCR-G protein complex can be simulated without specific nanobody; (2) the G protein activation ability of GPCR can be explained by molecular dynamics simulation. CONCLUSIONS: It is suggested that we could do molecular dynamics simulation of complex of GPCR-G protein without bound nanobody. Secondly, the simulation time reduced greatly by using homology modeling to generate complex of proteins. Thirdly, the molecular dynamics simulation will help us to know or even predict further protein-protein interactions.

Cytosolic PKM2 stabilizes mutant EGFR protein expression through regulating HSP90-EGFR association.

This corrects the article DOI: 10.1038/onc.2015.397.

Cytosolic PKM2 stabilizes mutant EGFR protein expression through regulating HSP90-EGFR association.

Secondary mutation of epidermal growth factor receptor (EGFR) resulting in drug resistance is one of the most critical issues in lung cancer therapy. Several drugs are being developed to overcome EGFR tyrosine kinase inhibitor (TKI) resistance. Here, we report that pyruvate kinase M2 (PKM2) stabilized mutant EGFR protein by direct interaction and sustained cell survival signaling in lung cancer cells. PKM2 silencing resulted in markedly reduced mutant EGFR expression in TKI-sensitive or -resistant human lung cancer cells, and in inhibition of tumor growth in their xenografts, concomitant with downregulation of EGFR-related signaling. Mechanistically, PKM2 directly interacted with mutant EGFR and heat-shock protein 90 (HSP90), and thus stabilized EGFR by maintaining its binding with HSP90 and co-chaperones. Stabilization of EGFR relied on dimeric PKM2, and the protein half-life of mutant EGFR decreased when PKM2 was forced into its tetramer form. Clinical levels of PKM2 positively correlated with mutant EGFR expression and with patient outcome. These results reveal a previously undescribed non-glycolysis function of PKM2 in the cytoplasm, which contribute to EGFR-dependent tumorigenesis and provide a novel strategy to overcome drug resistance to EGFR TKIs.

Enabling high solubility of ZnO in TiO2 by nanolamination of atomic layer deposition.

Zn-doped TiO2 nanotubes were fabricated by nanolaminated packing of alternating layers of TiO2 and ZnO by atomic layer deposition (ALD) using a polycarbonate (PC) membrane as a template. With 400 cycles of ALD, the nanotubes with a thickness of 28 nm and an outer diameter of 220 nm were obtained after removing the PC membrane by annealing at 450 °C. The doping concentration of ZnO in TiO2 depends on the precursor cycle ratio of ZnO to TiO2. With the precursor cycle ratio of ZnO : TiO2 at 0.04, a uniform bulk solubility of ∼8 at% is obtained, and the surface concentration of Zn is even higher, ∼16 at%. From the depth profiles measured by secondary ion mass spectrometry, Zn is uniformly distributed across the thickness, which is further confirmed by analyses of X-ray photoelectron spectroscopy, X-ray diffraction, and Raman spectroscopy. Additionally, from the transmission electron microscopic observation, the highly doped anatase TiO2 exhibits some regions of severe deformation that results in localized solid-state amorphization.

MTAP is an independent prognosis marker and the concordant loss of MTAP and p16 expression predicts short survival in non-small cell lung cancer patients.

OBJECTIVES: Methylthioadenosine phosphorylase (MTAP), a ubiquitously expressed protein, plays important roles in purine biosynthesis. Locating near to each other on chromosome 9p21-22, codeletion of the MTAP and p16(Ink4A) genes have been reported in non-small cell lung cancer (NSCLC). The aim of this study is to determine the respective prognostic value of MTAP and p16 by considering their correlation in NSCLC patients. MATERIALS AND METHODS: We analyzed MTAP and p16 protein expression by immunohistochemical staining on 99 NSCLC tissue microarray samples. The association between MTAP and p16 expression levels and prognosis were analyzed using the Kaplan-Meier method and Cox proportional hazards model for prognosis. RESULTS: Patients with a low MTAP expression level had poor overall survival (P = 0.010) and disease-free survival (P = 0.002). Low p16 expression indicated a trend toward poor overall survival (P = 0.138) and disease-free survival (P = 0.199). There was a significant positive correlation between MTAP and p16 expression levels (Spearman's ρ = 0.402, P < 0.001). By multivariate analyses, the MTAP expression level retained its independent prognostic power and p16 expression loss of the correlation with prognosis. Concordant loss of MTAP and p16 expression was observed in 24 out of 99 patients (24.2%). Patients with concordant loss of MTAP and p16 expression had the worst prognosis compared to patients with high expression of both markers. CONCLUSION: MTAP expression is an independent prognostic factor and has greater prognostic significance than p16 expression in NSCLC. Concordant loss of MTAP and p16 expression indicates poor outcomes in lung cancer patients.

Bacteria killing nanotechnology Bio-Kil effectively reduces bacterial burden in intensive care units.

A contaminated hospital environment has been identified as an important reservoir of pathogens causing healthcare-associated infections. This study is to evaluate the efficacy of bacteria killing nanotechnology Bio-Kil on reducing bacterial counts in an intensive care unit (ICU). Two single-bed rooms (S-19 and S-20) in the ICU were selected from 7 April to 27 May 2011. Ten sets of new textiles (pillow cases, bed sheets, duvet cover, and patient clothing) used by patients in the two single-bed rooms were provided by the sponsors. In the room S-20, the 10 sets of new textiles were washed with Bio-Kil; the room walls, ceiling, and air-conditioning filters were treated with Bio-Kil; and the surfaces of instruments (respirator, telephone, and computer) were covered with Bio-Kil-embedded silicon pads. Room S-19 served as the control. We compared the bacterial count on textiles and environment surfaces as well as air samples between the two rooms. A total of 1,364 samples from 22 different sites in each room were collected. The mean bacterial count on textiles and environmental surfaces in room S-20 was significantly lower than that in room S-19 (10.4 vs 49.6 colony-forming units [CFU]/100 cm(2); P < 0.001). Room S-20 had lower bacterial counts in air samples than room S-19 (33.4-37.6 vs 21.6-25.7 CFU/hour/plate; P < 0.001). The density of microbial isolations was significantly greater among patients admitted to room S-19 than those to room S-20 (9.15 vs 5.88 isolates per 100 patient-days, P < 0.05). Bio-Kil can significantly reduce bacterial burden in the environment of the ICU.

A bacteria injection scheme for in situ bioaugmentation.

This article presents an innovative design for inoculating the desired organisms to stratified geological layers at desired rates during in-situ bioaugmentation. The new delivery system consists of intermittent porous tubes connected in series with impermeable polyethylene tubes that run horizontally in each stratified layer of a contaminated aquifer. A bioaugmentation test using the new delivery system was conducted to inject an enriched culture of Escherichia coli (E. coli). Results of the test indicated that the distribution of E. coli through each porous tube was fairly uniform. A mathematical model previously developed to calculate the distribution of water flow through each porous tube was modified to calculate the distribution of E. coli. Geological layers often have different hydraulic conductivities. By controlling the permeability and the length of porous tubes placed in stratified layers, the new design provides a means to selectively deliver aqueous bacteria to various layers at desired rates according to aquifer heterogeneity.

A nutrient injection scheme for in situ bio-remediation.

Geological layers often have different hydraulic conductivities. This paper presents an innovative design for delivering aqueous substrates and nutrients to various stratified layers at desired rates during in-situ bio-stimulation. The new delivery system consists of intermittent porous tubes connected in series with impermeable polyethylene tubes that run horizontally in each stratified layer of a contaminated aquifer. Results of the tracer test indicated that the distribution of tritium through each porous tube was fairly uniform. A mathematical model was also developed to calculate the distribution of water flow through each porous tube. By controlling the permeability and the length of porous tubes placed in stratified layers, the new design provides a means to selectively deliver nutrients to various layers at desired rates according to aquifer heterogeneity.

Psychometric comparisons of three measures for assessing motor functions in preschoolers with intellectual disabilities.

BACKGROUND: Deficit in motor performance is common in children with intellectual disabilities (ID). A motor function measure with sound psychometric properties is indispensable for clinical and research use. The purpose of this study was to compare the psychometric properties of three commonly used clinical measures for assessing motor function in preschoolers with ID: the Bruininks-Oseretsky Test of Motor Proficiency-Second Edition, the Movement Assessment Battery for Children-Second Edition and the Peabody Developmental Motor Scale-Second Edition (PDMS-2). METHOD: One hundred and ninety-one children aged 3-6 years with ID were evaluated with the three measures at three time points: two baseline measurements with a 1-week interval before the intervention, and a follow-up measurement after 6 months of paediatric rehabilitation programme. One hundred and forty-one participants completed all of the assessments. The distribution (ceiling and floor effects) and reliability (internal consistency and test-retest reliability) of each measure were examined. Concurrent validity, predictive validity, and responsiveness were examined as well. RESULTS: All measures, except for the PDMS-2, had significant floor effects or ceiling effects at one or more time points. The three measures had good internal consistency (Cronbach α ≥ 0.86) and test-retest reliability (intraclass correlation coefficient ≥ 0.96). The Spearman ρ correlation coefficient for each pair of the three measures was ≥ 0.80, indicating high concurrent validity. The predictive validity of the three measures was satisfactory (Spearman ρ ≥ 0.52). The responsiveness of the three measures was moderate (0.47 ≤ effect size ≤ 0.74). The minimal detectable changes of the three measures were satisfactory. CONCLUSIONS: All three measures showed sufficient reliability, validity and responsiveness in preschoolers with ID, but the PDMS-2 is recommended for its superior psychometric properties.

Impact of solvent/detergent treatment of plasma on transfusion-relevant bacteria.

BACKGROUND: A solvent/detergent (S/D) treatment in a medical device has been developed for pathogen reduction of plasma for transfusion. Impact of S/D on bacterial growth and on the capacity of complement to kill bacteria has been investigated in this study. STUDY DESIGN AND METHODS: A pool of apheresis plasma from four donors was spiked with eight transfusion-relevant bacteria. Plasma was treated with 1% tri(n-butyl) phosphate and 1% Triton X-45 at 31°C for 90 min and then extracted by oil at 31°C for 70 min. Decomplemented plasma and Phosphate Buffer Saline were used as controls. Bacterial count was determined in samples taken immediately after spiking, or after S/D and oil treatment. Similar experiments were conducted using three individual recovered plasma donations. Bacteria growth inhibition tests were performed using discs soaked with plasma samples whether containing the S/D agents or not. RESULTS: The mean reduction factors of Escherichia coli, Pseudomonas aeruginosa, and Klebsiella pneumoniae due to complement during S/D treatment were >8·75, 4·71, and 4·18 log in pooled plasma and >7·42, 2·24 and >6·08 log in individual plasmas, respectively. Bacillus cereus and Bacillus subtilis were inactivated by S/D (>7·04 and 1·60 log in pooled, and >6·06 and 2·39 in individual plasmas, respectively). Staphylococcus aureus, Staphylococcus epidermidis and Enterobacter cloacae did not multiply during S/D treatment of plasma. Growth inhibition tests revealed an inhibition of three gram-negative bacteria by complement and all gram-positive by S/D. CONCLUSION: The S/D treatment of plasma does not alter the bactericidal activity of complement, and inactivates some gram-positive bacteria.

Interleukin 10 and residual kidney function are associated with risk of vascular calcification in patients undergoing peritoneal dialysis.

AIMS: Vascular calcification is a common complication among dialysis patients and its pathogenesis involves a variety of factors. The roles of pro-inflammatory cytokines and residual kidney function (RKF) in peritoneal dialysis (PD) patients with vascular calcification have not been investigated. MATERIALS AND METHODS: 157 stable PD patients were enrolled. All patients had plain X-ray film examination including chest (posterior-anterior view, CXR) and pelvis. Vascular calcification was interpreted as calcified deposit over aortic arch and linear calcification of pelvic arteries. Relevant biochemical data, pro-inflammatory markers, and PD-related factors were measured and collected. RESULTS: Vascular calcification prevalence in CXRs was higher than that in pelvis films (38.2% vs. 22.3%, p < 0.05). Patients with vascular calcification in CXR had higher incidence of calcification in pelvis films (p < 0.05). Only a minor portion (14.6%) had two calcification sites. Regression analysis revealed that age, PD duration, body mass index, and RKF were independent factors associated with vascular calcification in CXR. Age, diabetes, IL-10 and RKF were factors associated in pelvis films. Factors independently related to vascular calcification in both films were age, duration, diabetes, IL-10, and RKF. CONCLUSIONS: Besides traditional risk factors, IL-10 and RKF were important factors associated with vascular calcification in PD patients.

A chromatographically purified human TGF-ß1 fraction from virally inactivated platelet lysates.

BACKGROUND AND OBJECTIVES: TGF-ß1 exerts important physiological functions in osteogenesis and chondrogenesis and may be of therapeutic interest. The aim of this work was to develop a scalable purification process of TGF-ß1 from virally inactivated human platelets. STUDY DESIGN AND METHODS: Apheresis platelet concentrates (N=12) were solvent/detergent (S/D) treated (1% TnBP/1% Triton X-45; 31°C) and the resulting platelet lysates were clarified by oil extraction and centrifugation, then chromatographed on an anion-exchange DEAE-Sepharose Fast-Flow column equilibrated in a PBS buffer, pH 7.5. The column was washed to eliminate unbound proteins and the S/D agents. Bound proteins were eluted using a 1 M NaCl-PBS buffer pH 7.5 (DEAE-eluate). The content in TGF-ß1, PDGF-AB, VEGF, IGF-1, EGF, and b-FGF was measured by ELISA. Proteins, lipids, and S/D agents were assessed. Protein profile was determined by SDS-PAGE under reduced or non-reduced conditions. RESULTS: Most proteins, including albumin and immunoglobulins G, A, and M did not bind to the DEAE column as evidenced also by SDS-PAGE. Essentially all PDGF, VEGF, and IGF were in the breakthrough. The DEAE-eluate contained close to 60% of the TGF-ß1 at a mean concentration of about 102 ng/ml, whereas EGF, b-FGF were at about 0.72 and 0.18 ng/ml, respectively. The content in TnBP and Triton X-45 was <2 ppm. CONCLUSION: A fraction enriched in TGF-ß1 can be prepared from virally inactivated human platelet lysates using an easily scale process. Its interest in regenerative medicine and cell therapy will be evaluated in further studies.

Hospital-based surveillance and molecular epidemiology of rotavirus infection in Taiwan, 2005-2007.

To determine the distribution of rotavirus strains and facilitate vaccine policy decisions in Taiwan, active hospital-based gastroenteritis surveillance was conducted in three sentinel hospitals. From 1 January 2005 to 31 December 2007, a total of 3435 children less than 5 years old with gastroenteritis were enrolled. The presence of rotavirus was documented by enzyme immunoassay (EIA), and the G and P genotypes were determined by reverse transcription-polymerase chain reaction (RT-PCR) and sequencing methods. Results confirmed that 856 (25%) of these gastroenteritis admissions were EIA-positive for rotavirus and 448 (52%) of the rotavirus positive admissions were less than 2 years old. The most prevalent rotavirus genotypes were G1P[8] (40%), followed by strains G3P[8] (27%), and G9P[8] (17%). These data will help inform decisions as to whether rotavirus vaccine should be considered for inclusion into Taiwan's National Immunisation Programme.

Marsupialisation and strap muscle transposition laryngoplasty for vocal cysts with vocal fold atrophy.

OBJECTIVES: Vocal cysts with fold atrophy often result in more severe glottal incompetence than vocal cysts along during phonation. Although total excision or marsupialisation are reliable treatments for vocal fold cysts, any post-operative vocal deficit with significant glottal gap will need further treatment. This study aimed to evaluate the efficacy of combined treatment consisting of marsupialisation of the cyst immediately followed by strap muscle transposition laryngoplasty. METHOD: Under direct laryngomicroscopy, microscissors were used to make a disc-shaped incision encircling the equator of the cyst. After marsupialisation of the cyst, a simultaneous medialisation laryngoplasty with strap muscle transposition was performed. RESULTS: Seven patients with vocal cysts and marked vocal fold atrophy were included in the study. After surgery, subjective improvement in voice quality was reported by all patients. Patients' glottal incompetence and vocal performance were markedly improved. CONCLUSION: Marsupialisation is a simple and effective surgical technique for vocal fold cysts. For cases of vocal cysts with marked vocal fold atrophy, marsupialisation followed by medialisation laryngoplasty with strap muscle transposition may be considered.

Overexpression of cyclin E messenger ribonucleic acid in nasopharyngeal carcinoma correlates with poor prognosis.

AIMS: S-phase kinase-associated protein 2 is required for the degradation of p27 protein, which is a negative regulator of cyclin E/cyclin-dependent kinase 2 complex. The present study examined the expression of cyclin E, S-phase kinase-associated protein 2 and p27 protein in nasopharyngeal carcinoma. METHODS: Tissue from 35 cases of nasopharyngeal carcinoma and 10 normal nasopharyngeal tissue samples underwent reverse polymerase chain reaction to detect messenger ribonucleic acid. Immunohistochemical analysis was performed on 29 nasopharyngeal tissue samples in order to detect protein expression. RESULTS: Messenger ribonucleic acid expression in the nasopharyngeal carcinoma tissue samples analysed indicated a 1.75-fold change in the amount of S-phase kinase-associated protein 2, a 0.34-fold change in the amount of cyclin E and a 0.31-fold change in the amount of p27 protein, compared with positive controls. High levels of cyclin E significantly correlated with late-stage nasopharyngeal carcinoma (p = 0.009) and a poor overall survival (p = 0.010). Immunohistochemical analysis indicated positive expression of S-phase kinase-associated protein 2 in 16/29 nasopharyngeal tissue samples (55 per cent), of cyclin E in 13/29 samples (45 per cent) and of p27 protein in 17/29 (59 per cent) samples. CONCLUSIONS: Overexpression of cyclin E messenger ribonucleic acid showed an adverse prognostic significance, correlating with an advanced stage of nasopharyngeal carcinoma and a low overall survival rate.

A virally inactivated functional growth factor preparation from human platelet concentrates.

BACKGROUND: Human platelet growth factors (HPGF) are essential for tissue regeneration and may replace fetal bovine serum (FBS) in cell therapy. No method for the manufacture of standardized virally inactivated HPGF has been developed yet. STUDY DESIGN AND METHODS: Platelet concentrates (PC) were subjected to solvent/detergent (S/D) treatment (1% TnBP/1% Triton X-45), oil extraction, hydrophobic interaction chromatography and sterile filtration. Platelet-derived growth factor (PDGF)-AB, -BB and -AA, transforming growth factor-beta1 (TGF-beta1), epidermal growth factor (EGF), insulin-like growth factor-1 (IGF-1) and vascular endothelium growth factor (VEGF) were measured by ELISA. Composition in proteins and lipids was determined, protein profiles were obtained by SDS-PAGE, and TnBP and Triton X-45 were assessed by gas chromatography and high-performance liquid chromatography, respectively. Cell growth promoting activity of HPGF was evaluated by 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) assay using human embryonic kidney (HEK293A) fibroblast and Statens Seruminstitute rabbit corneal (SIRC) epithelial cell lines. RESULTS: The GF preparation contained a mean of 16.66, 2.04, 1.53, 72.19, 0.33, 48.59 and 0.44 ng/ml of PDGF-AB, -BB, -AA, TGF-beta1, EGF, IGF-1 and VEGF, respectively. The protein profile was typical of platelet releasates and had less than 2 p.p.m. of residual S/D agents. MTS assay of HEK293A and SIRC cultures showed that the GF preparation at 10% and 0.1% (v/v), respectively, could successfully replace 10% FBS for cell proliferation. Cell-stimulating activity of HPGF on HEK293A was over twice that of PC releasates. CONCLUSION: STANDARDIZED and functional virally inactivated HPGF can be prepared from human PC for possible applications in cell therapy and regenerative medicine.

Overexpression of beta2-microglobulin is associated with poor survival in patients with oral cavity squamous cell carcinoma and contributes to oral cancer cell migration and invasion.

beta2-Microglobulin (beta2M), a component of MHC class I molecules, is believed to be associated with tumour status in various cancers. In this study, we examined the expression of beta2M at different malignant stages of oral cavity squamous cell carcinoma (OCSCC). To determine the possible correlation between beta2M expression and various clinical characteristics, 256 samples from patients with OCSCC were evaluated by immunohistochemical staining. Strong beta2M expression was significantly correlated with a relatively advanced tumour stage (P<0.001), positive nodal status (P<0.001), and TNM stage (P<0.001). The cumulative 5-year survival rate was significantly correlated with a relatively advanced tumour stage (P<0.001), positive nodal status (P<0.001), TNM stage (P<0.001), and strong expression of beta2M (P<0.001). Thus, elevated beta2M expression is an indicator of poor survival (P<0.001). In addition, we extended our analysis of beta2M expression to the FaDu and SCC25 oral cancer cell lines. beta2-Microglobulin expression was positively correlated with cell migration and invasion in beta2M-overexpressing transfectants in Transwell chambers. The suppression of beta2M expression using small interfering RNA (siRNA) was sufficient to decrease cell migration and invasion in vitro. Taken together, our results suggest that beta2M expression in the tissues is associated with survival and may be involved in tumour progression and metastasis in OCSCC.