bluuebunny/crossref_metadata_2025_split huggingface dataset:
New pOp/LhG4 vectors for stringent glucocorticoid‐dependent transgene expression in Arabidopsis
Authors: Craft Judith , Samalova Marketa , Baroux Celia , Townley Helen , Martinez Alberto , Jepson Ian , Tsiantis Miltos , Moore Ian 
Published: March 2005

SummaryTo facilitate glucocorticoid‐inducible transgene expression from the pOp promoter in Arabidopsis the ligand‐binding domain of a rat glucocorticoid receptor (GR LBD) was fused to the amino terminus of the synthetic transcription factor LhG4 to generate LhGR‐N. Fusions bearing the GR LBD at other positions in LhG4 exhibited incomplete repression or inefficient induction. LhGR‐N was stringently repressed in the absence of exogenous glucocorticoid but was fully activated by addition of 2 μm dexamethasone which resulted in 1000‐fold increase in GUS reporter activity. Half maximal induction was achieved with 0.2 μm dexamethasone. Reporter transcripts were detectable within 2 h of dexamethasone application and peaked 4–10 h later. Neither LhGR‐N nor dexamethasone affected seedling development although ethanol retarded development when used as a solvent for dexamethasone. The efficiency of the pOp target promoter was improved 10‐ to 20‐fold by incorporating six copies of the ideal lac operator with sufficient inter‐operator spacing to allow simultaneous occupancy. Introduction of the TMV Ω sequence into the 5′UTR resulted in a further 10‐fold increase in dexamethasone‐inducible reporter activity and an increase in the induction factor to 104. Although promoters containing the TMV Ω sequence exhibited slightly increased basal expression levels in the absence of dexamethasone, stringent regulation of the cytokinin biosynthetic gene ipt was achieved with all promoters. Despite the severity of the induced ipt phenotypes, transcripts for the KNOX homoeodomain transcription factors BREVIPEDICELLUS and SHOOTMERISTEMLESS were not significantly increased within 48 h of dexamethasone application to seedlings.
Senescence‐associated vacuoles with intense proteolytic activity develop in leaves of Arabidopsis and soybean
Authors: Otegui Marisa S. , Noh Yoo‐Sun , Martínez Dana E. , Vila Petroff Martin G. , Andrew Staehelin L. , Amasino Richard M. , Guiamet Juan J. 
Published: March 2005

SummaryVacuolar compartments associated with leaf senescence and the subcellular localization of the senescence‐specific cysteine‐protease SAG12 (senescence‐associated gene 12) were studied using specific fluorescent markers, the expression of reporter genes, and the analysis of high‐pressure frozen/freeze‐substituted samples. Senescence‐associated vacuoles (SAVs) with intense proteolytic activity develop in the peripheral cytoplasm of mesophyll and guard cells in Arabidopsis and soybean. The vacuolar identity of these compartments was confirmed by immunolabeling with specific antibody markers. SAVs and the central vacuole differ in their acidity and tonoplast composition: SAVs are more acidic than the central vacuole and, whereas the tonoplast of central vacuoles is highly enriched in γ‐TIP (tonoplast intrinsic protein), the tonoplast of SAVs lacks this aquaporin. The expression of a SAG12‐GFP fusion protein in transgenic Arabidopsis plants shows that SAG12 localizes to SAVs. The analysis of ProSAG12:GUS transgenic plants indicates that SAG12 expression in senescing leaves is restricted to SAV‐containing cells, for example, mesophyll and guard cells. A homozygous sag12 Arabidopsis mutant develops SAVs and does not show any visually detectable phenotypical alteration during senescence, indicating that SAG12 is not required either for SAV formation or for progression of visual symptoms of senescence. The presence of two types of vacuoles in senescing leaves could provide different lytic compartments for the dismantling of specific cellular components. The possible origin and functions of SAVs during leaf senescence are discussed.
Large‐scale screening of Arabidopsis enhancer‐trap lines for seed germination‐associated genes
Authors: Liu Po‐Pu , Koizuka Nobuya , Homrichhausen Tanja M. , Hewitt Jessica R. , Martin Ruth C. , Nonogaki Hiroyuki 
Published: March 2005

SummaryEnhancer trap is a powerful approach for identifying tissue‐ and stage‐specific gene expression in plants and animals. For Arabidopsis research, β‐glucuronidase (GUS) enhancer‐trap lines have been created and successfully used to identify tissue‐specific gene expression in many plant organs. However, limited applications of these lines for seed germination research have been reported. This is probably due to the impermeability of the testa to the GUS substrate. By focusing on the stages between testa and endosperm rupture, we were able to circumvent the testa barrier to the GUS substrate and observe diverse tissue‐specific gene expression during germination sensu stricto. One hundred and twenty‐one positive subpools of 10 lines out of 1130 were isolated. Approximately 4500 plants from these subpools were grown in a greenhouse and one to seven individual plants exhibiting GUS expression in seeds were isolated for each subpool. This library of the Arabidopsis seed enhancer‐trap lines is an efficient tool for identifying seed germination‐associated genes. The individual lines from this library will be provided to the international seed biology research community. International collaboration to identify the trapped genes using genome‐walking PCR and to characterize the gene functions using knockout plants will significantly enhance our understanding of the molecular mechanisms of seed germination.
Endosomal proteases facilitate the fusion of endosomes with vacuoles at the final step of the endocytotic pathway
Authors: Yamada Kenji , Fuji Kentaro , Shimada Tomoo , Nishimura Mikio , Hara‐Nishimura Ikuko 
Published: March 2005

SummaryThe mechanism by which plasma membrane proteins are transported to vacuoles for degradation has not been well characterized in plants. To clarify how plasma membrane proteins are degraded, we monitored the endocytotic pathway in tobacco suspension‐cultured BY‐2 cells with a fluorescent endocytosis marker, FM4‐64. Because of the efficient and rapid delivery of endosomes to the vacuoles, endosomes were scarcely detectable. Interestingly, we found that E‐64d, an inhibitor of papain family proteases, caused the accumulation of a large number of endosomes in the cells under the sucrose‐starved condition. This result indicates that E‐64d attenuates the fusion of endosomes with vacuoles. We identified two papain homologues, which are localized in the endosomes, with a biotinylated inhibitor. We designated them as endosome‐localized papains (ENPs). Immunofluorescent analysis revealed that vacuolar sorting receptor, a marker of prevacuolar compartment (PVC), was localized in the endosomes. This result and their acidic nature show that the endosomes correspond to PVC. These results suggest that ENPs facilitate the final step in the vacuolar trafficking pathway under the sucrose‐starved condition. We further examined the effects of E‐64d on two transgenic Arabidopsis plants that constitutively express a fusion protein composed of green fluorescent protein (GFP) and a plasma membrane protein (GFP‐PIP2a or GFP‐LTI6b). GFP fluorescence was observed on the plasma membrane of root cells in these transgenic plants. Treatment with E‐64d induced the accumulation of GFP‐fluorescent endosomes and inhibited the degradation of these fusion proteins. No GFP fluorescence was observed in vacuoles in E‐64d‐treated transgenic plants. Taken together, these results suggest that endosomal proteases are required for the fusion of endosomes with vacuoles at the final step in the endocytotic pathway for degradation of plasma membrane proteins in plants.
The stability of the Arabidopsis transcriptome in transgenic plants expressing the marker genes nptII and uidA
Authors: Ouakfaoui Souad El , Miki Brian 
Published: March 2005

SummaryThe ATH1 Arabidopsis GeneChip from Affymetrix was used to search for transcriptome changes in Arabidopsis associated with the strong expression of transgenes regulated by constitutive promoters. The insertion and expression of the commonly used marker genes, uidA and nptII, did not induce changes to the expression patterns of the approximately 24 000 genes that were screened under optimal growth conditions and under physiological stress imposed by low temperatures. Approximately 8000 genes (35% of the Arabidopsis genome) underwent changes in gene expression in both wild‐type and transgenic plants under abiotic stresses such as salt, dehydration, cold, and heat. This study provides detailed information on the extent of non‐targeted or pleiotropic effects of transgenes on plants and shows that the transgenic and non‐transgenic plants were equivalent in their global patterns of transcription. This information may help to extend our understanding and interpretation of the principle of substantial equivalence which is used as a first step in the biosafety evaluation of transgenic crops.
Moisturizing effects of topical nicotinamide on atopic dry skin
Authors: Soma Yoshinao , Kashima Masato , Imaizumi Akiko , Takahama Hideto , Kawakami Tamihiro , Mizoguchi Masako 
Published: March 2005

AbstractBackground  Certain moisturizers can improve skin barrier function in atopic dermatitis. The effect of topical nicotinamide on atopic dry skin is unknown. We examined the effect of topical nicotinamide on atopic dry skin and compared the results with the effect of white petrolatum in a left–right comparison study.Methods  Twenty‐eight patients with atopic dermatitis, with symmetrical lesions of dry skin on both forearms, were enrolled, and were instructed to apply nicotinamide cream containing 2% nicotinamide on the left forearm and white petrolatum on the right forearm, twice daily over a 4‐ or 8‐week treatment period. Transepidermal water loss and stratum corneum hydration were measured by instrumental devices. The amount of the stratum corneum exfoliated by tape stripping (desquamation index) was determined by an image analyzer.Results  Nicotinamide significantly decreased transepidermal water loss, but white petrolatum did not show any significant effect. Both nicotinamide and white petrolatum increased stratum corneum hydration, but nicotinamide was significantly more effective than white petrolatum. The desquamation index was positively correlated with stratum corneum hydration at baseline and gradually increased in the nicotinamide group, but not in the white petrolatum group.Conclusions  Nicotinamide cream is a more effective moisturizer than white petrolatum on atopic dry skin, and may be used as a treatment adjunct in atopic dermatitis.
Effects of mycophenolate mofetil on donor‐specific antibody formation in renal transplantation
Authors: Lederer Stephan R , Friedrich Nicole , Banas Bernhard , von Welser Georg , Albert Ekkehard D , Sitter Thomas 
Published: March 2005

Abstract:  Background:  Mycophenolate mofetil (MMF) is a routinely used immunosuppressive agent that selectively blocks T‐ and B‐lymphocyte proliferation. The present study was designed to investigate the effects of this drug on human leukocyte(HLA) antibody production in general and donor‐specific antibody (DSA) formation in particular in 154 recipients of renal allografts.Patients and methods:  Renal allograft recipients were subdivided into three groups. Group 1 patients (n = 60) had received MMF since transplantation in combination with either cyclosporin A or tacrolimus and steroids. Group 2 patients (n = 29) had received an MMF‐free immunosuppressive regimen initially followed by addition of MMF some time later. Group 3 patients (n = 65) had received no MMF. Cyclosporin A or tacrolimus in combination with azathioprine and/or steroids were used for immunosuppression. DSA were demonstrated by enzyme‐linked immunosorbent assay (ELISA) for detection of panel‐reactive antibodies of HLA class I and II specificity.Results:  The HLA antibodies were found in 16.7%, 27.6% and 30.8% of transplant recipients in groups 1, 2 and 3, respectively. DSA were found in 8.3%, 17.2% and 20.0%, and non‐DSA in 10.0%, 20.7% and 24.6%, of patients in groups 1, 2 and 3, respectively.Conclusion:  The MMF reduces anti‐HLA class I and II antibody production and consequently DSA production in renal allograft recipients. Our data indicate this effect to be more pronounced in patients given MMF immediately after transplantation than in those in whom MMF is introduced some time later. The presence of DSA in the serum of renal allograft recipients is associated with poorer graft function (higher serum creatinine and more rejection episodes).
Rapamycin in patients with chronic renal allograft dysfunction
Authors: Wu Mai‐Szu , Chang Chiz‐Tzung , Hung Cheng‐Chieh 
Published: March 2005

Abstract:  Purpose:  Nephrotoxicity of calcineurin inhibitors (CNI) complicates the management of chronic renal allograft dysfunction. Rapamycin is a promising immunosuppressive agent free of nephrotoxicity. The effect of conversion from CNI to rapamycin in recipients with chronic allograft dysfunction is still unclear. We investigated the effect of rapamycin in patients with chronic allograft dysfunction.Methods:  We conducted a prospective study on kidney transplant recipients with chronic allograft dysfunction. The patients were under classic CNI, mycophenolate mofetil, and prednisolone triple therapy. They had progressive deterioration of the allograft function. They were converted from CNI to rapamycin directly and observed for 6 months. The CNI serum levels before the conversion were within recommended range. Allograft function, clinical features and adverse effects were evaluated before and after the rapamycin conversion.Results:  A total of 16 patients were enrolled. Six of them (37.5%) failed to have a smooth conversion because of deterioration of allograft function and intractable adverse effects. Ten patients (62.5%) went through the 6‐month observation period with improved graft function. The average reduction of serum creatinine was 27.7% (p < 0.001) in successful conversion. There were no significant differences on age, gender, lipid profile, sugar control, and rapamycin levels between successful and failed conversion. Anemia and diastolic blood pressure were significantly improved after successful conversion. The failed patients had a longer transplantation period (6.1 ± 4.1 vs. 11.2 ± 3.4 yr, p < 0.05). Two of them (12.5%) developed bacteria pneumonia. Self‐limited diarrhea developed in three patients (18.8%).Conclusion:  We suggested that conversion from CNI to rapamycin was beneficial in some kidney transplant recipients with chronic allograft dysfunction.
Arterial reconstruction with donor iliac vessels during pancreas transplantation: an intraoperative approach to arterial injury or inadequate flow
Authors: Moon Jang IL , Ciancio Gaetano , Burke George W 
Published: March 2005

Abstract: Background: Outcome of pancreas transplantation (PTX) has improved because of use of novel immunosuppression and advances in surgical technique. It is not uncommon for severe atherosclerosis in patients with type 1 insulin‐dependent diabetes mellitus or the presence of a previously transplanted organ to limit the options for vascular anastomosis. Herein we report the novel application of donor iliac arterial interposition grafts for arterial reconstruction in patients with severe iliac artery arteriosclerosis, and/or previous transplant who develop an arterial injury or stenosis during surgery.Methods: In five patients undergoing PTX, the external iliac artery was severely atherosclerotic and/or occupied by a previous vascular anastomosis. In four of the five patients, an arterial intimal dissection became apparent. The external iliac artery was excised and reconstructed with donor iliac artery interposition graft (end‐to‐end anastomosis). Pancreas or kidney was engrafted onto this arterial interposition graft (end‐to‐side anastomosis).Results: There was no operative morbidity related to this surgical approach. All grafts functioned well after transplantation. Distal lower extremities have no evidence of vascular insufficiency with mean follow‐up of 26 months (7–45 months).Conclusion: This surgical technique is an acceptable option during PTX for the patient with severe iliac artery arteriosclerosis with intraoperative intimal dissection or stenosis, or perhaps those with challenging arterial access.
Clinical utility of the Handicap Assessment and Resource Tool: An investigation of its use with the aged people in hospital
Authors: Barbara Anita , Whiteford Gail 
Published: March 2005

Aims:  This article presents the findings of a qualitative study investigating the clinical utility of the Handicap Assessment and Resource Tool (HART). Clinicians’ perceptions of the HART were explored in its use in acute hospital settings in a regional health service in New South Wales, Australia. Methods and Results:  The study, which used in‐depth, structured interviews with clinicians working in acute aged care, identified key themes, which point to an overall positive perception of the HART's clinical utility. The themes, identified through thematic analysis were: responses to structural features of the HART; processes of administration; perceived benefits for clients; and clinical utility in context. The article concludes with recommendations for the assessment of aged people in acute hospitals and the need for occupational therapists to continue analysing and establishing the clinical utility of the profession's assessment tools. Conclusion:  The findings of the study support the clinical utility of the HART when used with aged people in acute care settings. Overall, though participants found it time consuming to administer, they judged the HART to be a context sensitive tool that addressed the complex needs of their patients.
Using recombinant human thyroid‐stimulating hormone for the diagnosis of recurrent thyroid cancer
Authors: Blamey Stephen , Barraclough Bruce , Delbridge Leigh , Mernagh Paul , Standfield Lachlan , Weston Adele 
Published: March 2005

Background:  Recombinant human thyroid‐stimulating hormone (rhTSH) has been suggested as a diagnostic agent in the diagnosis of recurrent thyroid cancer, instead of the current practice of thyroid hormone (THT) withdrawal.Methods:  An evidenced‐based literature review was used as the basis for a cost‐utility, decision‐analytic model. Outcome measures were safety, efficacy (diagnostic performance and quality of life) and cost effectiveness of rhTSH.Results:  The literature search identified six comparative studies of rhTSH versus THT withdrawal. The most common adverse events associated with the use of rhTSH were headache (3.5−11.1%) and nausea (7.7−17%). When used as a diagnostic agent, the unadjusted sensitivity and specificity for rhTSH were 87% and 95%, respectively. Thus the use of rhTSH instead of THT withdrawal would result in a reduction in diagnostic accuracy, with 11% of patients’ disease status being misclassified. Use of rhTSH resulted in a higher quality of life in the period prior to diagnostic testing than THT withdrawal (P < 0.001). When the impact of diagnostic performance, patient compliance to follow‐up and modified quality of life were modelled over a 5 year time‐frame, the incremental cost per QALY of rhTSH relative to THT withdrawal was $51 344.42.Conclusions:  The use of rhTSH as a diagnostic agent appears to be safe but less diagnostically accurate and less cost‐effective (on whole of healthcare cost basis) when used in the follow‐up of patients with thyroid cancer who have had a previous negative radioiodine scan after thyroid hormone withdrawal.
Cost‐effective method for laparoscopic choledochotomy
Authors: Griniatsos John , Karvounis Evangelos , Arbuckle James , Isla Alberto‐Martinez 
Published: March 2005

Background:  Recent reports have noted that postoperative complications following open or laparoscopic choledochotomy for common bile duct (CBD) exploration are mainly related to the T‐tube presence, and that there has been no trend of decrease in the laparoscopic era. Laparoscopic endobiliary stent placement with primary closure of the CBD has been proposed as a safe and effective alternative to T‐tube placement.Methods:  Between January 1999 and January 2003, 53 consecutive patients suffering from proven choledocholithiasis underwent laparoscopic common bile exploration (LCBDE) via choledochotomy. In the early period, a T‐tube was placed at the end of the procedure (group A, n = 32) while, from June 2001 onwards, laparoscopic biliary stent placement and primary CBD closure were chosen as the drainage method (group B, n = 21).Results:  Six patients developed T‐tube‐related complications postoperatively. Univariate analysis revealed statistically significant lower morbidity rate and shorter postoperative hospital stay for the stent group. Although not statistically significant, a median saving of £780 per patient was observed in the stent group.Conclusion:  Biliary endoprosthesis placement following laparoscopic choledochotomy avoids the well‐known complications of a T‐tube, leading to a shorter postoperative hospital stay. The method is safe and effective and it should also be considered as cost‐effective compared to T‐tube placement. Further studies are required in order to document cost‐effectiveness of the method.
Effect of dexamethazone on hepatic vascular response in experimental sepsis
Authors: Tarim Mehmet Akin , Sayek Iskender , Erdemli Inci 
Published: March 2005

Background:  The aim of the present study was to evaluate the effect of dexamethasone‐pretreatment on the hepatic artery and portal vein of septic rats, which were generated by lipopolisaccarides (LPS) intraperitoneal injection.Method:  Thirty‐six albino Wistar rats were used and constructed as LPS (n = 12), control (n = 12), dexamethazone‐pretreatment (n = 6) and dexamethazone‐control (n = 6) groups. Hepatic artery and portal vein rings were excised and placed in Krebs−Henseleit solution. Vessel rings were contracted with phenylephrine adding to the organ chamber in cumulative doses. Then the contraction‐response curves were drawn.Results:  In the LPS group, phenylephrine evoked contractions were reduced in both hepatic artery and portal vein rings in comparison to the control group. In the dexamethasone‐control group, phenylephrine‐evoked contractions were increased but not significantly. Dexamethasone‐pretreatment increased the phenylephrine‐evoked contractions close to the values of control group for both types of rings.Conclusions:  Dexamethasone pretreatment corrected the vascular hyporeactivity to phenyleprine in isolated portal vein and hepatic artery rings prepared from the LPS treated rats in experimental sepsis. This might have occurred as a result of inhibition of inducible nitric oxide synthase expression.