bluuebunny/crossref_metadata_2025_split huggingface dataset:
WRKY46 functions as a transcriptional repressor of ALMT1, regulating aluminum‐induced malate secretion in Arabidopsis
Authors: Ding Zhong Jie , Yan Jing Ying , Xu Xiao Yan , Li Gui Xin , Zheng Shao Jian 
Published: October 2013

SummaryAluminum (Al) toxicity is the major limiting factor for crop production on acid soils, but the transcriptional regulation of Al tolerance genes is largely unknown. Here, we found that the expression of a WRKY domain‐containing transcription factor WRKY46 is inhibited by Al and expressed in root stele, whereas the expression of ALMT1, which encodes a malate efflux transporter, is induced by Al stress and spatially co‐localized with WRKY46 in root stele, indicating the possible interaction between WRKY46 and ALMT1 in Arabidopsis. Mutation of WRKY46 by T–DNA insertion leads to better root growth under Al stress, and lower root Al content compared with the wild‐type Col–0. The wrky46 mutant shows increased root malate secretion, which is consistent with the higher ALMT1 expression in the mutant. Transient expression analysis using truncated promoter of ALMT1 showed that ALMT1 expression can be inhibited by WRKY46 in tobacco leaves. The yeast one‐hybrid assay and ChIP‐qPCR analysis revealed that WRKY46 directly binds to ALMT1 promoter through specific W–boxes. Taken together, we demonstrated that WRKY46 is a negative regulator of ALMT1, mutation of WRKY46 leads to increased malate secretion and reduced Al accumulation in root apices, and thus confers higher Al resistance.
A deficiency in chloroplastic ferredoxin 2 facilitates effective photosynthetic capacity during long‐term high light acclimation in Arabidopsis thaliana
Authors: Liu Jun , Wang Peng , Liu Bing , Feng Dongru , Zhang Jie , Su Jianbin , Zhang Yang , Wang Jin–Fa , Wang Hong–Bin 
Published: October 2013

SummaryPhotosynthetic electron transport is the major energy source for cellular metabolism in plants, and also has the potential to generate excess reactive oxygen species that cause irreversible damage to photosynthetic apparatus under adverse conditions. Ferredoxins (Fds), as the electron‐distributing hub in the chloroplast, contribute to redox regulation and antioxidant defense. However, the steady‐state levels of photosynthetic Fd decrease in plants when they are exposed to environmental stress conditions. To understand the effect of Fd down‐regulation on plant growth, we characterized Arabidopsis thaliana plants lacking Fd2 (Fd2–KO) under long‐term high light (HL) conditions. Unexpectedly, Fd2–KO plants exhibited efficient photosynthetic capacity and stable thylakoid protein complexes. At the transcriptional level, photoprotection‐related genes were up‐regulated more in the mutant plants, suggesting that knockout Fd2 lines possess a relatively effective photo‐acclimatory responses involving enhanced plastid redox signaling. In contrast to the physiological characterization of Fd2–KO under short‐term HL, the plastoquinone pool returned to a relatively balanced redox state via elevated PGR5‐dependent cyclic electron flow during extended HL. fd2 pgr5 double mutant plants displayed severely impaired photosynthetic capacity under HL treatment, further supporting a role for PGR5 in adaptation to HL in the Fd2–KO plants. These results suggest potential benefits of reducing Fd levels in plants grown under long‐term HL conditions.
TCP3 interacts with R2R3‐MYB proteins, promotes flavonoid biosynthesis and negatively regulates the auxin response in Arabidopsis thaliana
Authors: Li Shutian , Zachgo Sabine 
Published: October 2013

SummaryTCP proteins belong to the plant‐specific bHLH transcription factor family, and function as key regulators of diverse developmental processes. Functional redundancy amongst family members and post‐transcriptional down‐regulation by miRJAW of several TCP genes complicate their functional characterization. Here, we explore the role of TCP3 by analyzing transgenic plants expressing miRJAW‐resistant mTCP3 and dominant‐negative TCP3SRDX. Seedlings and seeds of mTCP3 plants were found to hyper‐accumulate flavonols, anthocyanins and proanthocyanidins, whereas levels of proanthocyanidins were slightly reduced in TCP3SRDX plants. R2R3‐MYB proteins control not only early flavonoid biosynthetic steps but also activate late flavonoid biosynthetic genes by forming ternary R2R3‐MYB/bHLH/WD40 (MBW) complexes. TCP3 interacted in yeast with R2R3‐MYB proteins, which was further confirmed in planta using BiFC experiments. Yeast three‐hybrid assays revealed that TCP3 significantly strengthened the transcriptional activation capacity of R2R3‐MYBs bound by the bHLH protein TT8. Transcriptome analysis of mTCP3 and TCP3SRDX plants supported a role for TCP3 in enhancing flavonoid biosynthesis. Moreover, several auxin‐related developmental abnormalities were observed in mTCP3 plants. Transcriptome data coupled with studies of an auxin response reporter and auxin efflux carriers showed that TCP3 negatively modulates the auxin response, probably by compromising auxin transport capacity. Genetic experiments revealed that the chalcone synthase mutant tt4‐11 lacking flavonoid biosynthesis abrogated the auxin‐related defects caused by mTCP3. Together, these data suggest that TCP3 interactions with R2R3‐MYBs lead to enhanced flavonoid production, which further negatively modulates the auxin response.
The Fate of PrP GPI‐Anchor Signal Peptide is Modulated by P238S Pathogenic Mutation
Authors: Guizzunti Gianni , Zurzolo Chiara 
Published: October 2013

AbstractGlycosylphosphatidylinositol (GPI)‐anchored proteins are localized to the plasma membrane via a C‐terminally linked GPI anchor. The GPI anchor is added concomitantly to the cleavage of the carboxy‐terminal GPI‐anchor signal sequence, thereby causing the release of a C‐terminal hydrophobic peptide, whose fate has not yet been investigated. Here we followed the fate of the GPI‐attachment signal of the prion protein (PrP), a protein implicated in various types of transmissible neurodegenerative spongiform encephalopathies (TSE). The PrP GPI‐anchor signal sequence shows a remarkable and unusual degree of conservation across the species and contains two point mutations (M232R/T and P238S) that are responsible for genetic forms of prion disorders. We show that the PrP GPI‐anchor signal peptide (SP), but not the one from an unrelated GPI‐anchored protein (folate receptor), undergoes degradation via the proteasome. Moreover, the P238S point mutation partially protects the PrP GPI‐anchor SP from degradation. Our data provide the first attempt to address the fate of a GPI‐anchor SP and identify a role for the P238S mutation, suggesting the possibility that the PrP GPI‐anchor SP could play a role in neurodegenerative prion diseases.
External quality assessment in molecular immunohematology: the INSTAND proficiency test program
Authors: Flegel Willy A. , Chiosea Ion , Sachs Ulrich J. , Bein Gregor 
Published: October 2013

BackgroundGenotyping for red blood cell (RBC), platelet (PLT), and granulocyte antigens is a new tool for clinical pathology, transfusion medicine services, and blood banks. Proficiency in laboratory tests can be established by external quality assessments (EQAs), which are required for clinical application in many health care systems. There are few EQAs for molecular immunohematology.Study Design and MethodsWe analyzed the participation and pass rates in an EQA for RBC, PLT, and granulocyte antigens. This EQA was distributed by INSTAND, a large nonprofit provider of proficiency tests, twice per year since Fall 2006 as EQA Number 235 Immunohematology A (molecular diagnostic). The coordinators defined at the outset which alleles are mandatory for detection.ResultsThe number of participants steadily increased from 51 to 73 per proficiency by Fall 2012. More than 60 institutions utilized this EQA at least once a year. Approximately 80% of them participated in RBC, 68% in PLT, and 22% in granulocyte systems. With the exceptions of RHD (82%) and granulocytes (85%), pass rates exceeded 93%. While the pass rate increased for granulocyte and decreased for the ABO system, the pass rates for the other systems changed little over 6½ years.ConclusionsThe INSTAND proficiency test program was regularly used for EQA by many institutions, particularly in Central Europe. While the technical standards and pass rates in the participating laboratories were high, there has been little improvement in pass rates since 2006.
Development of a validated exam to assess physician transfusion medicine knowledge
Authors: Haspel Richard L. , Lin Yulia , Fisher Patrick , Ali Asma , Parks Eric ,   Biomedical Excellence for Safer Transfusion (BEST) Collaborative
Published: October 2013

BackgroundThere is evidence that physicians lack adequate transfusion medicine knowledge. To design needs‐based educational interventions to address this gap, a validated assessment tool is required. Previously published exams have not been created or validated using rigorous psychometric methods.Study Design and MethodsA modified Delphi method was used to achieve consensus regarding the essential knowledge and skills for physicians who transfuse blood products. To ensure content validity, members of an international organization of transfusion medicine experts (Biomedical Excellence for Safer Transfusion [BEST] Collaborative) participated in the exam design process. An exam, based on the most highly rated topics, was created and administered to individuals with a priori expected basic, intermediate, and expert levels of transfusion medicine knowledge. Rasch analysis, a psychometric technique used in high‐stakes medical licensure and board testing, was used to determine exam accuracy and precision.ResultsThirty‐six topics achieved ratings sufficient to be considered for inclusion in the exam (content validity index > 0.8). A 23‐question exam was administered to 49 individuals. Mean scores for individuals with expected basic, intermediate, and expert knowledge were 42, 62, and 82%, respectively (p < 0.0001). The exam achieved good fit with the Rasch model.ConclusionA validated exam has now been created to accurately assess transfusion medicine knowledge. This exam can be used to determine knowledge deficits and assist in the design of curricula to improve blood product utilization.
Treatment of refractory thrombotic thrombocytopenic purpura with N‐acetylcysteine: a case report
Authors: Li Gloria W. , Rambally Siayareh , Kamboj Jasmine , Reilly Sean , Moake Joel L. , Udden Mark M. , Mims Martha P. 
Published: October 2013

BackgroundThrombotic thrombocytopenic purpura (TTP) is a life‐threatening disease resulting in systemic microvascular thrombosis. The disease is caused by excessive platelet (PLT) adhesion to ultra‐large (UL) von Willebrand factor (VWF) multimers inadequately cleaved by the processing enzyme ADAMTS‐13. While many cases respond to plasma exchange performed with or without concurrent corticosteroids, treatment of the 10% to 20% of patients with refractory disease is difficult. Experimental studies demonstrating that N‐acetylcysteine (NAC) inhibits PLT binding to endothelial cell–secreted and anchored UL VWF multimers suggest that NAC may be useful in the treatment of TTP.Case ReportA 44‐year‐old woman presented with malaise, confusion, chest and abdominal pain, and transient visual loss. Laboratory results and peripheral blood smear were consistent with TTP. The patient was begun on plasma exchange and corticosteroid treatment, but after 10 days the PLT count was still less than 10.0 × 109/L and she developed a fever. Rituximab was initiated, but the patient's condition worsened and she became comatose. Antibiotics were initiated, but cultures remained sterile. After 3 days of coma and further clinical deterioration, treatment with NAC was begun. The patient received a loading dose of 150 mg/kg NAC intravenously (IV) over 1 hour. Within 18 hours the patient awakened abruptly and began communicating with medical personnel. Plasma exchange, corticosteroids, rituximab, and NAC infusion (150 mg/kg IV over 17 hr daily × 10 days) were continued and by Day 17 the PLT count was more than 50 × 109/L. The patient fully recovered and was discharged on Day 31.ConclusionThis is the first complete report of a TTP patient treated with NAC. NAC was a safe and effective supplementary treatment for refractory TTP in this patient.
An innovative biologic recycling process of leukoreduction filters to produce active human antimicrobial peptides
Authors: Vossier Ludivine , Leon Fanny , Bachelier Cécile , Marchandin Hélène , Lehmann Sylvain , Leonetti Jean‐Paul , Brodolin Konstantin , Coste Joliette , Fournier‐Wirth Chantal 
Published: October 2013

BackgroundHuman neutrophil peptides (HNPs) 1 to 3 are the major antimicrobial peptides of the azurophilic granules of neutrophils. They represent an important arm of the innate immune system. Their production by chemical synthesis and recombinant technologies is expensive and limited by technical constraints due to their composition and the presence of three disulfide bonds.Study Design and MethodsWe have developed an original approach based on the purification of the natural human defensins HNPs 1 to 3 from neutrophils trapped on leukoreduction filters used in blood processing. The purification of HNPs 1 to 3 from these filters is performed in two steps: extraction of HNPs 1 to 3 retained in the filters followed by their immunoprecipitation. Studies were performed to determine the stability of defensins in the filters stored at room temperature. The activity of HNPs 1 to 3 obtained by our rapid protocol was validated by determining minimal inhibitory concentrations (MICs) against six reference bacterial strains and 12 clinical isolates.ResultsThe human defensins HNPs 1 to 3 extracted from leukoreduction filters displayed high antimicrobial activity against tested strains, with MIC values between 0.12 and 1 μg/mL. Kinetics assays showed the appearance of activity 15 minutes after peptide addition. Moreover, we found that the HNPs 1 to 3 purified from leukoreduction filters that had been stored for 45 days at room temperature remained active.ConclusionLeukoreduction filters provide a rich and safe source of active human defensins HNPs 1 to 3. Moreover, the stability of the peptides in filters stored at room temperature allows envisaging a large‐scale development of the process.
The efficacy of cetirizine hydrochloride on the pruritus of cats with atopic dermatitis: a randomized, double‐blind, placebo‐controlled, crossover study
Authors: Wildermuth Kerstin , Zabel Sonja , Rosychuk Rod A. W. 
Published: October 2013

BackgroundVarious antihistamines have been used in the management of feline atopic dermatitis, with variable reported benefit. To date, there have been no randomized, double‐blind, placebo‐controlled, crossover clinical trials on the use of this drug class in cats.Hypothesis/ObjectivesTo evaluate the clinical efficacy of cetirizine hydrochloride for the control of pruritus and dermatitis in cats diagnosed with atopic dermatitis.MethodsIn this randomized, double‐blind, placebo‐controlled crossover clinical trial, 21 client‐owned cats diagnosed with mild to moderate nonseasonal atopic dermatitis were randomly assigned to two groups. Cats in each group received either 1 mg/kg cetirizine hydrochloride or placebo once daily per os for 28 days followed by a 14 day wash‐out period. Treatments were then crossed over, and cats received placebo or cetirizine hydrochloride for another 28 days. Owners marked a pruritus severity scale before inclusion in the study and weekly throughout the entire study period. Lesions were scored by the clinician using aCanineAtopicDermatitisExtent andSeverityIndex (CADESI)‐03 modified for the cat before enrolment and at day 28 of each treatment.ResultsNineteen cats completed the study. There were no statistically significant differences between treatment with cetirizine hydrochloride and placebo for modifiedCADESI‐03 or pruritus scores.Conclusion and clinical importanceThis study suggests that cetirizine hydrochloride cannot be recommended for the management of feline atopic dermatitis.
Transcription profile of chemokine receptors, cytokines and cytotoxic markers in peripheral blood of dogs with epitheliotropic cutaneous lymphoma
Authors: Chimura Naoki , Iio Aki , Ozaki Eiji , Mori Takashi , Ito Yusuke , Murayama Nobuo , Nagata Masahiko , Ide Kaori , Nishifuji Koji , Kamishina Hiroaki , Maeda Sadatoshi 
Published: October 2013

BackgroundMycosis fungoides (MF) is the most common form of canine epitheliotropic cutaneous lymphoma, which is characterized by the accumulation of neoplastic CD8+ T cells. Given that multifocal skin lesions are commonly seen in MF, neoplastic lymphocytes may actively migrate into the blood circulation.Hypothesis/ObjectivesCytotoxic T cells with a skin‐homing phenotype could be increased in the blood circulation of dogs with MF.AnimalsTen dogs with MF and 10 age‐matched healthy dogs were included.MethodsThe transcription levels of chemokine receptors, cytokines and cytotoxic markers in peripheral blood of dogs with MF were quantified by real‐time RT‐PCR.ResultsThe dogs with MF had lower transcription levels of chemokine receptors associated with skin homing (CCR4), epitheliotropism (CXCR3), lymph node homing (CCR7), a type‐1 cytokine (LT‐α) and cytotoxic markers (perforin and granzyme B) in the circulation than healthy control dogs (P < 0.05).Conclusions and clinical importanceThe present results suggest that the number of peripheral cytotoxic T cells with a skin‐homing phenotype could be decreased in the peripheral blood of dogs with MF, which might be due to the sequestration of cytotoxic T cells in the lesional skin.
An in vitro study to determine the minimal bactericidal concentration of sodium hypochlorite (bleach) required to inhibit meticillin‐resistant Staphylococcus pseudintermedius strains isolated from canine skin
Authors: Pariser Marlene , Gard Sharon , Gram Dunbar , Schmeitzel Lynn 
Published: October 2013

BackgroundMeticillin‐resistant Staphylococcus pseudintermedius (MRSP) is becoming a more common pathogen in animals treated in veterinary hospitals, raising concerns for transmission and possible contamination of the hospital itself. The identification of effective agents for the decontamination of veterinary patients and hospitals is crucial. Sodium hypochlorite (6.15%) or ‘bleach’ is an inexpensive and frequently used decontamination agent in human and veterinary hospitals and an adjunctive treatment for human patients. There are no published data on the minimal bactericidal concentration (MBC) of sodium hypochlorite (6.15%) for MRSP.Hypothesis/ObjectivesThe objective of this study was to determine the MBC of sodium hypochlorite (6.15%) against MRSP strains isolated from canine skin.MethodsIn this in vitro study, 12 canine skin isolates were obtained from the Antech Diagnostics microbiology department. Twofold serial dilutions of sodium hypochlorite (6.15%) were allowed to react with each of the MRSP strains for 15 min, followed by overnight incubation on agar plates. Colonies on each plate were counted. This process was repeated in triplicate.ResultsThe overall MBC for the canine skin‐isolated MRSP strains was 1:32, but most strains had an MBC between 1:64 and 1:128 dilution.Conclusions and clinical importanceThis study shows that sodium hypochlorite (6.15%) is an effective agent for decontamination at easily achievable concentrations.
Characterization of the complications associated with plasma exchange for thrombotic thrombocytopaenic purpura and related thrombotic microangiopathic anaemias: a single institution experience
Authors: McGuckin S. , Westwood J.‐P. , Webster H. , Collier D. , Leverett D. , Scully M. 
Published: October 2013

BackgroundPlasma exchange (PEX) is a life‐saving therapeutic procedure in patients with thrombotic thrombocytopaenic purpura (TTP) and other thrombotic microangiopathic anaemias (TMAs). However, it may be associated with significant complications, exacerbating the morbidity and mortality in this patient group.Study Design and MethodsWe reviewed all PEX procedures over a 72‐month period, following the exclusive introduction of solvent–detergent double viral‐inactivated plasma in high‐volume users, such as TTP, in the United Kingdom (UK). We documented allergic reactions to plasma, citrate reactions, complications relating to central venous access insertion and venous thrombotic events (VTE) in 155 patient episodes and >2000 PEX procedures.ResultsThe overall complication rate was low. Allergic plasma reactions occurred in 6·45% of the cohort with only one episode of acute anaphylaxis. Similarly, VTEs were 6·45%, not significantly greater than in medical patients receiving thromboprophylaxis, despite added potential risk factors in TTP. Citrate reactions were the most frequent complication documented, but toxicity was significantly reduced by administration of further calcium infusions during the PEX procedure. There were no serious central line infections and no catheter thrombosis.ConclusionOur data confirms that PEX continues to be a life‐saving procedure in the acute TTP setting and, the procedure was not associated with an increased mortality and limited morbidity.
Compliance with the current 12‐month deferral for male‐to‐male sex in Australia
Authors: Seed C. R. , Lucky T. T. , Waller D. , Wand H. , Lee J. F. , Wroth S. , McDonald A. , Pink J. , Wilson D. P. , Keller A. J. 
Published: October 2013

Background and ObjectivesIn Australia since 2000, donors are deferred for 12 months since last male‐to‐male sexual contact. There is no estimate of the prevalence of non‐compliance (i.e. failure to disclose a risk during the predonation interview which would lead to deferral) with the policy in Australia; however, published studies elsewhere indicate a range of 0·8–11%. We investigated the rate of, timing and motivation for non‐compliance.Materials and MethodsA nationally representative sample of donors who had made a recent donation negative for transfusion‐transmissible infection testing was surveyed using an anonymous, online instrument. Non‐compliance was considered as a ‘yes’ response to the current screening question. Non‐compliers were requested to define the timing of the last sexual contact relevant to their most recent donation. Univariate and multivariate regression analyses were used to define factors associated with non‐compliance.ResultsOf 14 476 responses from male donors, 34 (0·23%, 95% CI: 0·16–0·33%) were non‐compliant of whom 24 (0·17%, 95% CI: 0·11–0·25%) had contact within 6 months of donation. Factors significantly associated with non‐compliance included: multiple sexual partners, history of injecting drug use, perception of a lack of privacy during interview and preference for a computer‐based questionnaire.ConclusionOur study confirms high compliance (>99·7%) to the 12‐month deferral for male‐to‐male sex in Australia providing reassuring evidence for the efficacy of the screening question. Issues of ‘privacy’ and ‘discomfort’ associated with disclosure suggest the use of validated audio computer‐assisted structured interview as a possible option for improving compliance with the donor questionnaire.