Extended basis sets for Pb to be used in conjunction with the Stevens et al. (denoted by SKBJ) and the Stuttgart effective core potentials (ECPs) have been optimized. An assessment of different ab ...initio and density functional procedures showed that all methods based on the use of the SKBJ ECP exaggerate the Pb
+ binding enthalpies, which are systematically 35–50 kJ mol
−1 larger than those obtained when the Stuttgart ECP is employed. The G96LYP/Stutt method yields values in close agreement with those obtained at the QCISD(T)/Stutt and CCSD(T)/Stutt levels of theory, but at a significantly lower cost. Also the B3LYP/Stutt, the BLYP/Stutt and the MP2/Stutt approaches provide a good compromise between accuracy and computational cost for the calculation of binding enthalpies.
Departments of 1 Cellular and Molecular Physiology and 2 Medicine, Milton S. Hershey Medical Center, Pennsylvania State University, Hershey; 3 Weis Center for Research, Geisinger Medical Center, ...Danville, Pennsylvania; and 4 Department of Internal Medicine (Cardiovascular Division), University of Virginia Health Sciences Center, Charlottesville, Virginia
Submitted 27 June 2005
; accepted in final form 26 September 2005
Messenger RNA levels of phospholemman (PLM), a member of the FXYD family of small single-span membrane proteins with putative ion-transport regulatory properties, were increased in postmyocardial infarction (MI) rat myocytes. We tested the hypothesis that the previously observed reduction in Na + -K + -ATPase activity in MI rat myocytes was due to PLM overexpression. In rat hearts harvested 3 and 7 days post-MI, PLM protein expression was increased by two- and fourfold, respectively. To simulate increased PLM expression post-MI, PLM was overexpressed in normal adult rat myocytes by adenovirus-mediated gene transfer. PLM overexpression did not affect the relative level of phosphorylation on serine 68 of PLM. Na + -K + -ATPase activity was measured as ouabain-sensitive Na + -K + pump current (Ip). Compared with control myocytes overexpressing green fluorescent protein alone, Ip measured in myocytes overexpressing PLM was significantly ( P < 0.0001) lower at similar membrane voltages, pipette Na + (Na + pip ) and extracellular K + (K + o ) concentrations. From 70 to +60 mV, neither Na + pip nor K + o required to attain half-maximal Ip was significantly different between control and PLM myocytes. This phenotype of decreased V max without appreciable changes in K m for Na + and K + in PLM-overexpressed myocytes was similar to that observed in MI rat myocytes. Inhibition of Ip by PLM overexpression was not due to decreased Na + -K + -ATPase expression because there were no changes in either protein or messenger RNA levels of either 1 - or 2 -isoforms of Na + -K + -ATPase. In native rat cardiac myocytes, PLM coimmunoprecipitated with -subunits of Na + -K + -ATPase. Inhibition of Na + -K + -ATPase by PLM overexpression, in addition to previously reported decrease in Na + -K + -ATPase expression, may explain altered V max but not K m of Na + -K + -ATPase in postinfarction rat myocytes.
primary cardiac myocyte culture; patch clamp; ion transport; Western blots
Address for reprint requests and other correspondence: J. Y. Cheung, Dept. of Cellular and Molecular Physiology, Milton S. Hershey Medical Center MC-H166, Hershey, PA 17033 (e-mail: jyc1{at}psu.edu )
This study explored a strategy to convert agricultural and forestry residues into microbial lipid, which could be further transformed into biodiesel. Among the 250 yeast strains screened for xylose ...assimilating capacity, eight oleaginous yeasts were selected by Sudan Black B test. The lipid content of these 8 strains was determined by soxhlet extraction method. One strain (T216) was found to produce lipids up to 36.6%, and it was identified as Rhodotorula glutinis. The optimal fermentation conditions were obtained as follows: glucose as carbon source 100 g/L; yeast extract and peptone as nitrogen sources at, respectively, 8 and 3 g/L; initial pH of 5.0; inoculation volume of 5%; temperature at 28 degree C, shaking speed of 180 r/min, cultivated for 96 h. Under these conditions, R. glutinis accumulated lipids up to 49.25% on a cellular biomass basis and the corresponding lipid productivity reached 14.66 g/L. Experiments with a 5-L bioreactor under the optimal culture conditions showed that R. glutinis accumulated lipids up to 60.69%, resulting in 23.41 g/L in lipid productivity. More encouraging results were observed for the lipid production with alternative carbon sources. Corn stalk and Populus euramevicana leaves hydrolysate could be used to substitute glucose. Chemical analysis indicated that biodiesel obtained by transesterification possessed similar composition to that from vegetable oil, one of the widely used feedstock for biodiesel.
Forest ecosystems in the Northern Hemisphere function as carbon (C) sinks for atmospheric carbon dioxide; however, the magnitude, location, and cause of the sinks remain uncertain. A number of field ...measurements of forest biomass and systematic national forest inventories in Japan make it possible to quantify the C sinks and their distribution. Allometric relationships between forest biomass and stem volume were obtained for the major forest types in Japan from 945 sets of direct field measurements across the country. These relationships were used to estimate the changes in C accumulations of aboveground biomass and total living biomass from 1947 to 1995 from the national forest inventories of 1947, 1956, 1961, 1965, 1975, 1980, 1985, 1990, and 1995. The results showed that the C accumulations have significantly increased during the last 50 years. The C density (C stock per hectare) and total C stock of aboveground biomass increased from 27.6 Mg C/ha and 611.7 Tg C in 1947 to 43.2 Mg C/ha and 1027.7 Tg C in 1995, respectively, and those of total living biomass increased from 33.9 Mg C/ha and 751.8 Tg C in 1947 to 53.6 Mg C/ha and 1274.8 Tg C in 1995. These increases were remarkable during 1976–1995, with a net increase of 5.6 Mg C/ha and 369 Tg C for the C density and total living biomass. These results suggest that Japan's forest vegetation is a significant C sink. In the past 20 years, living vegetation has sequestered 18.5 Tg C annually, 14.6 Tg C of which was accumulated in aboveground biomass. The total C sink for the whole forest sector (including nonliving biomass) of Japan was estimated as 36 Tg C/yr if using the net change ratio of nonliving biomass C to living biomass C derived from the United States and Europe. On the basis of average C sink per hectare, Japan's forests have a higher sequestration rate (0.77 Mg C ha−1 yr−1) than the average of the other northern countries (0.14–0.19 Mg C ha−1 yr−1). The expansion and regrowth of planted forests are two major causes for this increased C uptake; planted forests contribute ∼80% of the total C sink in Japan. The suitable oceanic climate for fast forest growth and effective forest management practice may be the principal factors for such a large sink.
: Background/Aim: Recent data have shown that the clinical outcome of hepatitis C virus (HCV) infection may be influenced by the host genetic factor. The aim of this study was to investigate whether ...particular human leukocytes antigen (HLA) molecules are associated with the susceptibility to HCV infection in the Korean population.
Methods: One hundred and thirty‐seven patients with chronic HCV infection and 206 normal individuals were examined for HLA class I and II molecules.
Results: In class I antigens, the frequencies of HLA‐A3 (relative risk (RR)=3.5, P<0.04), HLA‐B35 (RR=2.0, P<0.03), and HLA‐B46 (RR=2.5, P<0.02) significantly increased in chronic HCV carriers compared with the controls. The frequencies of DRB1*0803, DQB1*0601 and DQB1*0604 were significantly higher in chronic HCV carriers than in controls (RR=2.5, P<0.005; RR=1.8, P<0.05; RR=1.9, P<0.04, respectively). On the other hand, the frequencies of DRB1*0301, DQA1*0501 and DQB1*0201 were significantly lower in chronic HCV carriers than in normal controls (RR=0.2, P<0.03; RR=0.4, P<0.004; RR=0.5, P<0.02, respectively). The haplotype DRB1*0803−DQB1*0601 significantly increased (RR=2.5, P<0.02) while the DQA1*0501−DQB1*0201 significantly decreased (RR=0.2, P<0.03) in chronic HCV carriers compared with normal controls. In stratification analysis to investigate the interrelationships among the associated alleles, DRB1*0803 and DQB1*0601 were associated with HLA‐B46, particularly in patients with chronic HCV carriers.
Conclusions: These results suggest that particular HLA alleles may have an influence on chronic HCV infection as a host genetic factor in the Korean population.
This study is conducted to evaluate the feasibility of percutaneous transhepatic gallbladder drainage prior to laparoscopic cholecystectomy for the treatment of gallbladder empyema. We also determine ...the sonographic findings, causative organism, clinical signs and symptoms, laboratory data, associated underlying medical disorders and the complications related to both cholecystostomy and laparoscopic cholecystectomy.
One hundred and forty-five cases of gallbladder empyema were included in this study which was composed of 80 males and 65 females, aged 22-94 years with a mean age of 71-years. All patients underwent percutaneous transhepatic gallbladder drainage under ultrasound and fluoroscopic guidance, and laparoscopic cholecystectomy was carried out thereafter. We analyzed the clinical presentations (signs, symptoms, laboratory and ultrasonographic findings, concomitant medical disorders), causative organisms and the complications related to percutaneous cholecystostomy and laparoscopic cholecystectomy.
Percutaneous transhepatic gallbladder drainage was performed successfully in all patients within 48 hours after clinical diagnosis of acute cholecystitis. Complications related to percutaneous transhepatic gallbladder drainage were bile leakage after tract dilatation noted in 2 patients (1.4%), and 20 (14%) patients had pain at the puncture site which radiated to the right shoulder during the procedure, but resolved spontaneously within an hour later. On admission, 102 (70%) patients presented as right upper quadrant pain, 39 (27%) as epigastric pain, 90 (62%) as fever, 108 (74%) patients had leukocytosis, and 33 (22.7%) patients were septic. AST and ALT were elevated in 57% and 51% of patients, respectively. Alkaline phosphatase was elevated in 56% of patients, and 34% of those patients had combined common bile duct stones. Gallbladder stones were documented in 135 (93%) patients, while the remaining 10 (7%) cases were acalculous. Five (3.4%) patients had combined gallbladder adenocarcinoma, 7 (4.8%) had liver abscess, while 13 (9%) had biliary pancreatitis. The ultrasonographic findings included gallbladder distension (93%), wall thickening (90%), pericholecystic fluid accumulation (15%), intraluminal sludge or stone (93%) and intraluminal air (13.9%). Bile culture were positive in 83% of the cases and showed gram-negative bacteria in 75%, gram-positive in 30%, anaerobes in 7%, while no growth in the remaining 17% of the cases. The common pathogens were Escherichia coli (57%), Enterococcus (27%), Klebsiella pneumonia (18%), Morganella morganii (7.6%), Pseudomonas aeruginosa (4.1%) and Salmonella (0.7%). The total postoperative complication rate was 17%, which included wound infection, bleeding, subhepatic abscess, cystic duct stump leak, common bile duct injury and pneumonia. Postoperative mortality was 2.6%. Conversion rate to open cholecystectomy was 27%. Clinical conditions improved within 48 hours after cholecystostomy in 93% of patients. Time interval between cholecystostomy and elective cholecystectomy was 2-21 days with a mean of 4 days. Total hospital stay was 5-38 days (mean: 11 days).
Percutaneous transhepatic gallbladder drainage is a safe and effective procedure for the initial management of gallbladder empyema. We highly recommend this preoperative drainage procedure in patient with sepsis, and for those high-risk patients such as old age and with underlying medical illnesses. This procedure can stabilized the patient so that an appropriate therapeutic planning can be achieved.
Products of the pts operon of Escherichia coli have multiple physiological roles such as sugar transport, and the operon is controlled by two promoters, P0 and P1. Expression of the pts P0 promoter ...that is increased during growth in the presence of glucose is also activated by cAMP receptor protein·cAMP. Based on the existence of a sequence that has a high similarity with the known Mlc binding site in the promoter, the effects of the Mlc protein on the pts P0 promoter expression were studied. In vivo transcription assays using wild type and mlc-negative E. coli strains grown in the presence and absence of glucose indicate that Mlc negatively regulates expression of the P0 promoter, and Mlc-dependent repression is relieved by glucose in the growth medium. In vitro transcription assay using purified recombinant Mlc showed that Mlc repressed transcription from the P0 but did not affect the activity of the P1. DNase I footprinting experiments revealed that a Mlc binding site was located around +1 to +25 of the promoter and that Mlc inhibited the binding of RNA polymerase to the P0 promoter. Cells overexpressing Mlc showed a very slow fermentation rate compared with the wild type when grown in the presence of various phosphoenolpyruvate-carbohydrate phosphotransferase system sugars but few differences in the presence of non-phosphoenolpyruvate-carbohydrate phosphotransferase system sugars except maltose. These results suggest that the pts operon is one of major targets for the negative regulation by Mlc, and thus Mlc regulates the utilization of various sugars as well as glucose in E. coli. The possibility that the inducer of Mlc may not be sugar or its derivative but an unknown factor is proposed to explain the Mlc induction mechanism by various sugars.
Abstract
Background
Immune system has been known to influence the prognosis of breast cancer (BC). However, the relationship between immune modulating factor (PD-L1) and tumor infiltrating lymphocyte ...(TIL) profiles in breast cancer has yet to be revealed according to breast cancer subtypes. In addition, the effects of circulating cytokines on TILs have not been addressed.
Patients and methods
We investigated the relationship between the profiles of TILs and PD-L1 expression of the primary tumor tissue by immunohistochemistry with clinical outcomes in 253 patients who underwent surgery for early breast cancer at National Cancer Center from January 2001 to December 2005. Besides, the serum cytokines including IL-10, IL-18, IL-6, IFN-g, and TGF-β1 were measured at diagnosis. Clinical data including hormone receptors status, HER2 expression, disease free survival (DFS), and overall survival (OS) were collected.
Results
Median age of patients was 49 years (range, 32-74) and median follow-up was 8.5 years. One hundred eighty five (73.1%) patients had hormone receptor (HR) positive and 101 (39.9%) patients had node positive BC. CD8+ TILs were more abundant in low PD-L1 expressed tumor (P = 0.027), though there was no association between FOXP3+ TILs and PD-L1 expression (P = 0.585). A total number of TILs was higher in HR negative compared with HR positive BC (P = 0.061) and the expression of PD-L1 was more frequent in HR positive BC (P<0.001). In HR negative BC, there was a trend of longer DFS in patients with higher CD8+ TILs and low PD-L1 expression (P = 0.097). However, such association was not detected in HR positive BC patients. Among serum cytokines we examined, the higher levels of IL-18 were significantly associated with shorter DFS in HR negative BC (P = 0.006). In HR negative BC, higher CD8+ TILs with low PD-L1 expression and lower IL-18 were significantly related with better clinical outcomes when adjusted with other clinical factors (DFS, P = 0.032; OS, P = 0.048).
Conclusions
Lower PD-L1 expression in breast tumor was associated with higher CD8+ lymphocyte infiltration. Especially in HR negative BC, increasing CD8+ TILs with lower PD-L1 expression and lower serum IL-18 level were good prognostic factors. Further validation will be needed to establish the role of immune profiles in BC patients.
Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P5-01-12.
Measurements of thermal O+ ion densities, field-aligned velocities, and fluxes from the Thermal Ion Dynamics Experiment (TIDE) on Polar obtained near 5000 km altitude over the Southern Hemisphere are ...compared with auroral images from the Ultraviolet Imager (UVI). Three passes were selected for analysis in this paper based on data availability from the TIDE and UVI instruments. Results indicate upward O+ flows in the cleft region but downward O+ flows in the polar cap region. Also, the O+ ion density follows a decreasing trend from the poleward side of the cusp region into the nightside aurora region. The magnitude of the downward O+ parallel velocities increases from dayside to nightside across the polar cap boundary. The upflows tend to occur over or near auroral forms, while the downflows are seen in relatively dark regions, such as the polar cap. These results are consistent with a cleft ion fountain source for the polar cap O+ ions. In the nightside polar cap, the results indicate a transition from downward to upflowing field-aligned O+ ions near boundaries of bright auroral arcs.
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