bluuebunny/crossref_metadata_2025_split huggingface dataset:
Metabolic network reconstruction and flux variability analysis of storage synthesis in developing oilseed rape (Brassica napus L.) embryos
Authors: Hay Jordan , Schwender Jörg 
Published: April 2011

SummaryComputational simulation of large‐scale biochemical networks can be used to analyze and predict the metabolic behavior of an organism, such as a developing seed. Based on the biochemical literature, pathways databases and decision rules defining reaction directionality we reconstructed bna572, a stoichiometric metabolic network model representing Brassica napus seed storage metabolism. In the highly compartmentalized network about 25% of the 572 reactions are transport reactions interconnecting nine subcellular compartments and the environment. According to known physiological capabilities of developing B. napus embryos, four nutritional conditions were defined to simulate heterotrophy or photoheterotrophy, each in combination with the availability of inorganic nitrogen (ammonia, nitrate) or amino acids as nitrogen sources. Based on mathematical linear optimization the optimal solution space was comprehensively explored by flux variability analysis, thereby identifying for each reaction the range of flux values allowable under optimality. The range and variability of flux values was then categorized into flux variability types. Across the four nutritional conditions, approximately 13% of the reactions have variable flux values and 10–11% are substitutable (can be inactive), both indicating metabolic redundancy given, for example, by isoenzymes, subcellular compartmentalization or the presence of alternative pathways. About one‐third of the reactions are never used and are associated with pathways that are suboptimal for storage synthesis. Fifty‐seven reactions change flux variability type among the different nutritional conditions, indicating their function in metabolic adjustments. This predictive modeling framework allows analysis and quantitative exploration of storage metabolism of a developing B. napus oilseed.
Perceived information needs and non‐adherence: evidence from Greek patients with hypertension
Authors: Stavropoulou Charitini 
Published: April 2011

AbstractBackground  The role of information on patients’ decision to non‐adhere is important, yet not well explored.Objective  To identify differences between perceived information needs for hypertension and medication to treat it, to explore the information channels used by patients and to test what type of information is more important to adhere to medication.Design, setting and participants  A questionnaire study was designed and conducted by telephone in the Centre for the Treatment of Hypertension in Athens, Greece, among 743 individuals.Main variables studied  The main variables included perceived information needs, information channels, non‐adherence to medication and socio‐demographic characteristics.Main outcome measures  Non‐adherence to medication was measured using the Morisky scale.Results  Patients reported feeling better informed about hypertension (90%) than medication to treat it (80%). The doctor remains the dominant information source, while the Media and magazines on health issues were reported more frequently than the family and the pharmacist. Feeling well informed about medication for hypertension was a predictor of better adherence. Other determinants of adherence were the use of the Internet and the Media.Discussion  The results confirm the importance of patients leaving the consultation feeling well informed about their medication as this improves adherence. They also show that the use of the Internet and the Media can be beneficial for adherence.Conclusions  Given the restricted time the doctor can usually spend with the patient, it is important to know that more emphasis on the information regarding medication is important.
Which anti‐platelet therapies might be beneficial in xenotransplantation?
Authors: Schmelzle Moritz , Cowan Peter J. , Robson Simon C. 
Published: April 2011

Schmelzle M, Cowan PJ, Robson SC. Which anti‐platelet therapies might be beneficial in xenotransplantation? Xenotransplantation 2011; 18: 79–87. © 2011 John Wiley & Sons A/S.Abstract:  Xenotransplantation could provide an unlimited and elective supply of grafts, once mechanisms of graft loss and vascular injury are better understood. The development of α‐1,3‐galactosyltransferase gene‐knockout (GalT‐KO) swine with the removal of a dominant xeno‐antigen has been an important advance; however, delayed xenograft and acute vascular reaction in GalT‐KO animals persist. These occur, at least in part, because of humoral reactions that result in vascular injury. Intrinsic molecular incompatibilities in the regulation of blood clotting and extracellular nucleotide homeostasis between discordant species may also predispose to thrombophilia within the vasculature of xenografts. Although limited benefits have been achieved with currently available pharmacological anti‐thrombotics and anti‐coagulants, the highly complex mechanisms of platelet activation and thrombosis in xenograft rejection also require potent immunosuppressive interventions. We will focus on recent thromboregulatory approaches while elucidating appropriate anti‐platelet mechanisms. We will discuss potential benefits of additional anti‐thrombotic interventions that are possible in transgenic swine and review recent developments in pharmacological anti‐platelet therapy.
Versatile co‐expression of graft‐protective proteins using 2A‐linked cassettes
Authors: Fisicaro Nella , Londrigan Sarah L. , Brady Jamie L. , Salvaris Evelyn , Nottle Mark B. , O’Connell Philip J. , Robson Simon C. , d’Apice Anthony J. F. , Lew Andrew M. , Cowan Peter J. 
Published: April 2011

Fisicaro N, Londrigan SL, Brady JL, Salvaris E, Nottle MB, O’Connell PJ, Robson SC, d’Apice AJF, Lew AM, Cowan PJ. Versatile co‐expression of graft‐protective proteins using 2A‐linked cassettes. Xenotransplantation 2011; 18: 121–130. © 2011 John Wiley & Sons A/S.Abstract:  Background:  Expression of multiple graft‐protective proteins targeted to different locations (i.e., intracellular, cell surface, and secreted) has become an increasingly important goal in xenotransplantation. The 2A “ribosome skip” signal is used as a linker to enable transgene co‐expression, but some studies have shown that post‐translational modification and trafficking of 2A‐linked proteins may be adversely affected depending on their position relative to 2A. We tested whether several relevant proteins, subject to a range of processing and localization mechanisms, could be efficiently co‐expressed using the 2A system.Methods:  Six expression cassettes were constructed, each containing up to four 2A‐linked open reading frames, encoding combinations of human CD55, thrombomodulin (TBM), CD39, CTLA4‐Ig and hygromycin resistance. Each linker incorporated a furin cleavage site to remove the carboxy‐terminal extension that remains on upstream proteins after 2A processing. The cassettes were used to produce vectors for transfection, adenoviral transduction and transgenesis. Expression was detected by flow cytometry and/or Western blotting.Results:  All proteins were expressed in the appropriate location following transient transfection of COS‐7 cells, irrespective of the number of linked genes. The percentage of stable transfectants expressing a linked gene was increased 10‐fold (from 4–5% to 58–67%) by incorporating the hygromycin resistance gene into the cassette. Stable transfection of transgenic GalT KO pig fibroblasts with a hygromycin‐ TBM‐CD39 construct resulted in surface expression of both TBM and CD39 by the majority of hygromycin‐resistant cells. Expression was maintained after flow cytometric sorting and expansion. Adenoviral transduction of NIT‐1 mouse insulinoma cells with a TBM‐CD39 construct resulted in strong expression of both genes on the cell surface. Mice transgenic for 3‐gene (CD55‐ TBM‐CD39) or 4‐gene (CD55‐ TBM‐CTLA4Ig‐CD39) constructs expressed all genes except CD55.Conclusions:  These results confirm the versatility of the 2A system, and demonstrate that careful construct design can minimize potential problems with post‐translational modification and trafficking. In addition, incorporation of a selection marker into the 2A‐linked chain can dramatically increase the proportion of stable transfectants expressing proteins of interest. This provides a powerful method for the rapid modification of existing genetically modified pigs.
Absence of Gal epitope prolongs survival of swine lungs in an ex vivo model of hyperacute rejection
Authors: Nguyen Bao‐Ngoc H. , Azimzadeh Agnes M. , Schroeder Carsten , Buddensick Thomas , Zhang Tianshu , Laaris Amal , Cochrane Megan , Schuurman Henk‐Jan , Sachs David H. , Allan James S. , Pierson III Richard N. 
Published: April 2011

Nguyen B‐NH, Azimzadeh AM, Schroeder C, Buddensick T, Zhang T, Laaris A, Cochrane M, Schuurman H‐J, Sachs DH, Allan JS, Pierson RN. Absence of Gal epitope prolongs survival of swine lungs in an ex vivo model of hyperacute rejection. Xenotransplantation 2011; 18: 94–107. © 2011 John Wiley & Sons A/S.Abstract:  Background:  Galactosyl transferase gene knock‐out (GalTKO) swine offer a unique tool to evaluate the role of the Gal antigen in xenogenic lung hyperacute rejection.Methods:  We perfused GalTKO miniature swine lungs with human blood. Results were compared with those from previous studies using wild‐type and human decay‐accelerating factor‐transgenic (hDAF+/+) pig lungs.Results:  GalTKO lungs survived 132 ± 52 min compared to 10 ± 9 min for wild‐type lungs (P = 0.001) and 45 ± 60 min for hDAF+/+ lungs (P = 0.18). GalTKO lungs displayed stable physiologic flow and pulmonary vascular resistance (PVR) until shortly before graft demise, similar to autologous perfusion, and unlike wild‐type or hDAF+/+ lungs. Early (15 and 60 min) complement (C3a) and platelet activation and intrapulmonary platelet deposition were significantly diminished in GalTKO lungs relative to wild‐type or hDAF+/+ lungs. However, GalTKO lungs adsorbed cytotoxic anti‐non‐Gal antibody and elaborated high levels of thrombin; their demise was associated with increased PVR, capillary congestion, intravascular thrombi and strong CD41 deposition not seen at earlier time points.Conclusions:  In summary, GalTKO lungs are substantially protected from injury but, in addition to anti‐non‐Gal antibody and complement, platelet adhesion and non‐physiologic intravascular coagulation contribute to Gal‐independent lung injury mechanisms.
Identification of soluble and membrane‐bound isoforms of porcine tumor necrosis factor receptor 2
Authors: Uribe‐Herranz Mireia , Casinghino Sandra R. , Bosch‐Presegué Laia , Fodor William L. , Costa Cristina 
Published: April 2011

Uribe‐Herranz M, Casinghino SR, Bosch‐Presegué L, Fodor WL, Costa C. Identification of soluble and membrane‐bound isoforms of porcine tumor necrosis factor receptor 2. Xenotransplantation 2011; 18: 131–146. © 2011 John Wiley & Sons A/S.Abstract:  Background:  TNF and its receptors TNF‐Receptor 1 (TNFR1, CD120a) and TNF‐Receptor 2 (TNFR2, CD120b) have been implicated in the rejection of transplanted cells and organs. Although pig TNFR1 (pTNFR1) is known to mediate the effects of human TNF in a xenogeneic setting, it is unclear whether pig TNFR2 (pTNFR2) could contribute to xenograft rejection.Methods:  We have cloned the cDNA of various pTNFR2 variants by reverse transcription–polymerase chain reaction (RT‐PCR) and rapid amplification of cDNA ends. We have characterized the predicted proteins with bioinformatic tools and conducted expression, affinity, and functional studies to investigate their roles.Results:  We have identified four isoforms of pTNFR2: one comprising the four cysteine‐rich domains (CRD) conserved between species, a shorter variant (pTNFR2ΔE7‐10) encoding for a soluble isoform, another with only three CRD due to the lack of exon 4 (pTNFR2ΔE4), and a fourth variant containing both modifications. Accordingly, multiple mRNA transcripts were observed by northern blotting. Quantitative RT‐PCR determined high pTNFR2 expression in lung and immune cells and detected the two alternative splicings in all cells/tissues examined. The full receptor was moderately expressed on the surface of pig cells such as porcine aortic endothelial cells and PK‐15 and was regulated by TNF. On the contrary, the membrane‐bound pTNFR2ΔE4 was located only intracellularly. Plasmon resonance studies showed that pTNFR2 binds pig and human TNFα with high affinity, but pTNFR2ΔE4 interacts poorly with pig TNFα and does not bind to the human cytokine. Moreover, pull‐down experiments with the two recombinant soluble isoforms consistently demonstrated that the two bound together and soluble pTNFR2ΔE4 was able to modulate the TNF inhibitory activity of pTNFR2‐GST in a cell‐based assay.Conclusion:  The pTNFR2 may participate in the process of xenograft rejection and other related events, as well as be used in soluble form to block TNF in this setting. In addition, we have discovered other pTNFR2 isoforms that may affect the pig immune responses and have an impact on rejection of xenografts.
Plastidic isoprenoid biosynthesis in tomato: physiological and molecular analysis in genotypes resistant and sensitive to drought stress
Authors: Loyola J. , Verdugo I. , González E. , Casaretto J. A. , Ruiz‐Lara S. 
Published: April 2011

AbstractIsoprenoid compounds synthesised in the plastids are involved in plant response to water deficit. The functionality of the biosynthetic pathway of these compounds under drought stress has been analysed at the physiological and molecular levels in two related species of tomato (Solanum chilense and Solanum lycopersicum) that differ in their tolerance to abiotic challenge. Expression analysis of the genes encoding enzymes of these pathways (DXS, IPI, GGPPS, PSY1, NCED and HPT1) in plants at different RWC values shows significant differences for only GGPPS and HPT1, with higher expression in the tolerant S. chilense. Chlorophyll, carotenoids, α‐tocopherol and ABA content was also determined in both species under different drought conditions. In agreement with HPT1 transcriptional activity, higher α‐tocopherol content was observed in S. chilense than in S. lycopersicum, which correlates with a lower degree of lipoperoxidation in the former species. These results suggest that, in addition to lower stomatal conductance, α‐tocopherol biosynthesis is part of the adaptation mechanisms of S. chilense to adverse environmental conditions.
Sulfur dioxide derivatives depress L‐type calcium channel in rat cardiomyocytes
Authors: Zhang Rong‐Yuan , Du Jun‐Bao , Sun Yan , Chen Stella , Tsai Hao‐Jan , Yuan Lan , Li Lin , Tang Chao‐Shu , Jin Hong‐Fang 
Published: April 2011

Summary1. Sulfur dioxide (SO2) has recently been found to have various biological effects on the cardiovascular system. The present study was designed to explore the effects of SO2 derivatives on the L‐type calcium current (I Ca, L) in isolated rat ventricular cardiomyocytes.2. A Langendorf system was used to dissociate single ventricular cells. SO2 derivatives from 5 to 1000 μmol/L were incubated with cardiomyocytes. The whole‐cell patch‐clamp technique was used to record I Ca, L. The effect of SO2 derivatives on intracellular calcium concentration ([Ca2+]i) was detected by confocal microscopy.3. Concentrations of 5 or 10 μmol/L SO2 derivatives could not change I Ca, L evoked by a single pulse from −40 to 0 mV for 200 ms in rat ventricular cardiomyocytes; however, 50, 100, 500 or 1000 μmol/L SO2 derivatives could depress the peak amplitudes of calcium currents in 6 min, and the I Ca, L was attenuated by 13.19%, 16.59%, 21.23% and 24.72%, respectively, as compared with corresponding controls (P < 0.05). The 50, 100, 500 or 1000 μmol/L SO2 derivatives also depressed the peak I‐V curves, without altering the reversal potential and the voltage dependence of the peak I Ca, L. Therefore, 1000 μmol/L SO2 derivatives could reduce [Ca2+]i in cardiomyocytes.4. The results of the present study suggest that SO2 derivatives can depress I Ca, L in cardiomyocytes, which might have a protective effect in cardiovascular diseases.
Co‐administration of meso 2,3‐dimercaptosuccinic acid monoesters reduces arsenic concentration and oxidative stress in gallium arsenide exposed rats
Authors: Flora Swaran JS , Bhatt Kapil , Dwivedi Nidhi , Pachauri Vidhu , Kushwah Pramod K 
Published: April 2011

Summary1. Gallium arsenide (GaAs), a semiconductor, exerts toxicity as a result of its constitutive moieties; that is, gallium and arsenic that becomes dissociated after exposure. The present study focuses on reducing arsenic concentration from the target organs using monoesters of meso 2,3‐dimercaptosuccinic acid (DMSA) either individually or in combination.2. Animals were exposed to GaAs (0.0014 mol/kg, orally for 8 weeks) and then treated with monoisoamyl DMSA (MiADMSA), monocyclohexyl DMSA (MchDMSA) or monomethyl DMSA (MmDMSA) either individually (0.3 mmol/kg, orally) or in combination (0.15 mmol/kg each, orally) for five consecutive days.3. GaAs exposure significantly inhibited blood δ‐aminolevulinic acid dehydrogenase (ALAD), suggesting alterations in the heme synthesis pathway. Whereas a significant increase in blood, liver and kidney reactive oxygen species accompanied by an increase in lipid peroxidation points to the involvement of oxidative stress in GaAs toxicity.4. GaAs also significantly disturbed glutathione metabolism. Hepatic and renal catalase activity decreased significantly, whereas hepatic and renal superoxide dismutase activity, as well as serum transaminases activity, showed marginal increase. Treatment with MiADMSA in combination with MchDMSA showed better therapeutic efficacy compared with other treatments in the aforementioned variables.5. Co‐administration of MiADMSA with MchDMSA provided better therapeutic effects, including reduction of arsenic burden, compared with all other treatments.
Ferritin/alanine aminotransferase ratio as a possible marker for predicting the prognosis of acute liver injury
Authors: Ozawa Eisuke , Abiru Seigo , Nagaoka Shinya , Yano Koji , Komori Atsumasa , Migita Kiyoshi , Yatsuhashi Hiroshi , Taura Naota , Ichikawa Tatsuki , Ishibashi Hiromi , Nakao Kazuhiko 
Published: April 2011

AbstractBackground and Aims:  Serum levels of ferritin and heme oxygenase (HO)‐1 are both markers of macrophage activation. We evaluated simple markers for predicting the prognosis of severe acute liver injury in which macrophage activation plays an important role.Methods:  Subjects comprised 114 patients with acute liver injury, admitted to the liver unit of Nagasaki Medical Center between January 2001 and September 2010. Subjects included 11 patients with fulminant hepatic failure (FHF), 82 patients with ordinary acute hepatitis (AH), and 21 patients with severe‐form AH (AHS). We determined serum levels of ferritin, HO‐1 and other biochemical makers, and analyzed relationships between clinical outcomes of patients and each of these parameters alone and in combination.Results:  Median serum ferritin levels were significantly higher in FHF (25 900 ng/mL) and AHS (3060 ng/mL) than in AH (700 ng/mL; P < 0.01 each). Median HO‐1 levels were also significantly higher in FHF (123 ng/mL) and AHS (51 ng/mL) than in AH (19 ng/mL; P < 0.01 each). Similarly, median ferritin/alanine aminotransferase (F/A) ratio was significantly higher in FHF (6.7) than in AHS (1.6, P < 0.05) or AH (0.5, P < 0.01). Among the 11 FHF patients, three recovered, seven died and one underwent liver transplantation. The ability of F/A ratio to distinguish non‐survivors from survivors was analyzed using receiver operating characteristics curves. A cut‐off level of 3.12 provided high sensitivity (87.5%) and specificity (81.2%).Conclusion:  These results suggest that F/A ratio offer a quick and simple marker for predicting the prognosis of acute liver injury.
Association of a single nucleotide polymorphism near the interleukin‐28B gene with response to hepatitis C therapy in Asian patients
Authors: Sinn Dong H , Kim Yu J , Lee Seung‐Tae , Gwak Geum‐Youn , Choi Moon S , Lee Joon H , Koh Kwang C , Yoo Byung C , Paik Seung W 
Published: April 2011

AbstractBackground and Aims:  A single nucleotide polymorphism near the interleukin‐28B (IL28B) gene has been shown to predict hepatitis C virus (HCV) treatment response. We aim to determine the role of the IL28B genotype in Asian patients.Methods:  A total of 118 patients (all Korean, 55 patients with genotype 1 infection and 63 patients with genotype 2 infection) were consecutively enrolled and analyzed.Results:  The sustained virological response (SVR) rate was 74% (87/118), while 26 patients (22%) relapsed and five patients were non‐responders (4%). For rs8099917, the frequencies of major homozygotes (TT), heterozygotes (GT), and minor homozygotes (GG) were 0.85, 0.14 and 0.01, respectively. Of the 55 patients with HCV genotype 1 infection, the SVR rate was 67% and 44% (P = 0.19) and the non‐response rate was 2% and 22% (P = 0.015) for the major allele and minor or hetero allele, respectively. Of the 63 patients with HCV genotype 2 infection, the SVR rate was 80% and 100% (P = 0.13) and the non‐response rate was 4% and 0% (P = 0.55) for major allele and hetero allele, respectively.Conclusions:  The IL28B genotype may help identify non‐responding patients in HCV genotype 1, but not in HCV genotype 2. Because of the high frequency of favorable alleles and the low frequency of non‐response, the IL28B polymorphism may play a smaller role in Asian patients.
Efficacy of therapy for advanced hepatocellular carcinoma: Intra‐arterial 5‐fluorouracil and subcutaneous interferon with image‐guided radiation
Authors: Chuma Makoto , Taguchi Hiroshi , Yamamoto Yoshiya , Shimizu Shinichi , Nakanishi Mitsuru , Ogawa Koji , Sho Takuya , Horimoto Hiromasa , Kobayashi Tomoe , Nakai Masato , Terashita Katsumi , Sakuhara Yusuke , Abo Daisuke , Tsukuda Yoko , Tsunematsu Seiji , Hige Shuhei , Kato Mototsugu , Shirato Hiroki , Asaka Masahiro 
Published: April 2011

AbstractBackground and Aim:  To evaluate the efficacy of intra‐arterial 5‐fluorouracil (5‐FU) and subcutaneous interferon (IFN) combined with image‐guided radiation therapy (IGRT) in advanced hepatocellular carcinoma (HCC) with portal vein tumor thrombosis (PVTT).Methods:  Twenty HCC patients with PVTT were treated with 5‐FU and IFN combined with image‐guided radiation therapy (IGRT) (IGRT group), and as controls, 20 patients with PVTT were treated with 5‐FU and IFN alone (non‐IGRT group). Overall survival (OS) time, response rates, time to progression (TTP) and safety were compared across groups.Results:  Complete response (CR), partial response (PR), stable disease (SD) and progressive disease (PD) of PVTT were 5%, 55%, 40% and 0% in the IGRT group and 0%, 30%, 35% and 35%, in the non‐IGRT group, respectively. CR, PR, SD, and PD of the whole tumor were 0%, 35%, 45% and 20% in the IGRT group and 0%, 30%, 35% and 35%, in the non‐IGRT group, respectively. Overall median survival was significantly longer in the IGRT group (12.0 months 95% confidence interval [CI], 9.3–17.6 months) than in the non‐IGRT group (9.1 months [95% CI, 5.5–11.1 months]) (P = 0.041). TTP was significantly longer in the IGRT group (6.9 months [95% CI, 5.6–10.2 months]) than in the non‐IGRT group (4.0 months [95% CI, 3.3–6.4 months]) (P = 0.034).Conclusions:  The response rates, median OS time and TTP in patients with advanced HCC with PVTT who received this novel combination therapy of intra‐arterial 5‐FU and subcutaneous IFN with IGRT are encouraging, and this combination therapy warrants further investigation.