bluuebunny/crossref_metadata_2025_split huggingface dataset:
Characterization of FAB1 phosphatidylinositol kinases in Arabidopsis pollen tube growth and fertilization
Authors: Serrazina Susana , Dias Fernando Vaz , Malhó Rui 
Published: May 2014

Summary

In yeast and animal cells, phosphatidylinositol‐3‐monophosphate 5‐kinases produce phosphatidylinositol (3,5)‐bisphosphate (PtdIns(3,5)P2) and have been implicated in endomembrane trafficking and pH control in the vacuole. In plants, PtdIns(3,5)P2 is synthesized by the Fab1 family, four orthologs of which exist in Arabidopsis: FAB1A and FAB1B, both from the PIKfyve/Fab1 family; FAB1C and FAB1D, both without a PIKfyve domain and of unclear role.

Using a reverse genetics and cell biology approach, we investigated the function of the Arabidopsis genes encoding FAB1B and FAB1D, both highly expressed in pollen.

Pollen viability, germination and tube morphology were not significantly affected in homozygous mutant plants. In vivo, mutant pollen fertilized ovules leading to normal seeds and siliques. The same result was obtained when mutant ovules were fertilized with wild‐type pollen. Double mutant pollen for the two genes was able to fertilize and develop plants no different from the wild‐type.

At the cellular level, fab1b and fab1d pollen tubes were found to exhibit perturbations in membrane recycling, vacuolar acidification and decreased production of reactive oxygen species (ROS). Subcellular imaging of FAB1B‐GFP revealed that the protein localized to the endomembrane compartment, whereas FAB1D‐GFP localized mostly to the cytosol and sperm cells. These results were discussed considering possible complementary roles of FAB1B and FAB1D.
A Comparison of Fentanyl and Flurbiprofen Axetil on Serum VEGF‐C, TNF‐α, and IL‐1ß Concentrations in Women Undergoing Surgery for Breast Cancer
Authors: Wen Yiyun , Wang Mingde , Yang Jinfeng , Wang Yichun , Sun Huiping , Zhao Jianghong , Liu Weizhen , Zhou Zhengyu , Deng Hongwu , Castillo‐Pedraza Catalina , Zhang Yi , Candiotti Keith A. 
Published: May 2014

AbstractBackgroundVascular endothelial growth factor‐C (VEGF‐C), tumor necrosis factor‐α (TNF‐α), and interleukin‐1ß(IL‐1ß) have been shown to be associated with the recurrence and metastasis of breast cancer after surgery. This study tested the hypothesis that patients undergoing surgery for breast cancer, who received postoperative analgesia with flurbiprofen axetil combined with small doses of fentanyl (FA), exhibited reduced levels of VEGF‐C, TNF‐α, and IL‐1ß compared with those patients receiving fentanyl alone (F).MethodForty‐women with primary breast cancer undergoing a modified radical mastectomy were randomized to receive postoperative analgesia with flurbiprofen axetil combined with fentanyl or fentanyl alone. Venous blood was sampled before anesthesia, at the end of surgery, and at 48 hours after surgery, and the serum was analyzed. The primary endpoint was changes in the VEGF‐C concentrations in serum.ResultsGroup FA patients reported similar analgesic effects as group F patients at 2, 24, and 48 hours. At 48 hours, mean postoperative concentrations of VEGF‐C in group F patients were higher than in group FA patients, 730.9 versus. 354.1 pg/mL (P = 0.003), respectively. The mean postoperative concentrations of TNF‐α in group F patients were also higher compared with group FA patients 27.1 vs. 15.8 pg/mL (P = 0.005). Finally, the mean postoperative concentrations of IL‐1ß in group F were also significantly higher than in group FA 497.5 vs. 197.7 pg/mL (P = 0.001).ConclusionIn patients undergoing a mastectomy, postoperative analgesia with flurbiprofen axetil, combined with fentanyl, were associated with decreases in serum concentrations of VEGF‐C, TNF‐α, and IL‐1ß compared with patients receiving doses of only fentanyl.
Genetic and phenotypic characterization of Botrytis calthae
Authors: Plesken C. , Westrich L.‐D. , Hahn M. 
Published: May 2014

Botrytis calthae is a necrotrophic plant pathogen, closely related to the ubiquitous broad host range fungus Botrytis cinerea, but highly host specific. Botrytis isolates from lesions of Caltha palustris grown at different locations were classified with genetic markers as either B. calthae or Botrytis pseudocinerea, or less frequently as B. cinerea. A PCR‐based identification of B. calthae was developed. Seven haplotypes of B. calthae could be distinguished. Compared to B. cinerea, mycelium growth of B. calthae was similar, but conidiation less abundant, and sclerotia formation was only partially repressed by light. Conidia of B. calthae germinated more slowly, and showed a highly acidic optimum (pH 2·5) compared to B. cinerea conidia (pH 5·3). All B. calthae isolates were sensitive to common anti‐Botrytis fungicides, but showed partial resistance to the succinate dehydrogenase inhibitors boscalid, fluopyram and carboxin. Infection experiments revealed a weak capability of B. calthae to induce necrotic lesions on plants that are hosts for B. cinerea. On C. palustris leaves, B. calthae induced similar lesions to B. cinerea. These data provide a basis for comparative molecular investigation of the physiology and host specificity of B. calthae and closely related Botrytis species.
Aligning mouse models of asthma to human endotypes of disease
Authors: Martin Rebecca A. , Hodgkins Samantha R. , Dixon Anne E. , Poynter Matthew E. 
Published: May 2014

AbstractSubstantial gains in understanding the pathophysiologic mechanisms underlying asthma have been made using preclinical mouse models. However, because asthma is a complex, heterogeneous syndrome that is rarely due to a single allergen and that often presents in the absence of atopy, few of the promising therapeutics that demonstrated effectiveness in mouse models have translated into new treatments for patients. This has resulted in an urgent need to characterize T helper (Th)2‐low, non‐eosinophilic subsets of asthma, to study models that are resistant to conventional treatments such as corticosteroids and to develop therapies targeting patients with severe disease. Classifying asthma based on underlying pathophysiologic mechanisms, known as endotyping, offers a stratified approach for the development of new therapies for asthma. In preclinical research, new models of asthma are being utilized that more closely resemble the clinical features of different asthma endotypes, including the presence of interleukin‐17 and aTh17response, a biomarker of severe disease. These models utilize more physiologically relevant sensitizing agents, exacerbating factors and allergens, as well as incorporate time points that better reflect the natural history and chronicity of clinical asthma. Importantly, some models better represent non‐classical asthma endotypes that facilitate the study of non‐Th2‐driven pathology and resemble the complex nature of clinical asthma, including corticosteroid resistance. Placing mouse asthma models into the context of human asthma endotypes will afford a more relevant approach to the understanding of pathophysiological mechanisms of disease that will afford the development of new therapies for those asthmatics that remain difficult to treat.
Global assessment of exposure to faecal contamination through drinking water based on a systematic review
Authors: Bain Robert , Cronk Ryan , Hossain Rifat , Bonjour Sophie , Onda Kyle , Wright Jim , Yang Hong , Slaymaker Tom , Hunter Paul , Prüss‐Ustün Annette , Bartram Jamie 
Published: May 2014

AbstractObjectivesTo estimate exposure to faecal contamination through drinking water as indicated by levels of Escherichia coli (E. coli) or thermotolerant coliform (TTC) in water sources.MethodsWe estimated coverage of different types of drinking water source based on household surveys and censuses using multilevel modelling. Coverage data were combined with water quality studies that assessed E. coli or TTC including those identified by a systematic review (n = 345). Predictive models for the presence and level of contamination of drinking water sources were developed using random effects logistic regression and selected covariates. We assessed sensitivity of estimated exposure to study quality, indicator bacteria and separately considered nationally randomised surveys.ResultsWe estimate that 1.8 billion people globally use a source of drinking water which suffers from faecal contamination, of these 1.1 billion drink water that is of at least ‘moderate’ risk (>10 E. coli or TTC per 100 ml). Data from nationally randomised studies suggest that 10% of improved sources may be ‘high’ risk, containing at least 100 E. coli or TTC per 100 ml. Drinking water is found to be more often contaminated in rural areas (41%, CI: 31%–51%) than in urban areas (12%, CI: 8–18%), and contamination is most prevalent in Africa (53%, CI: 42%–63%) and South‐East Asia (35%, CI: 24%–45%). Estimates were not sensitive to the exclusion of low quality studies or restriction to studies reporting E. coli.ConclusionsMicrobial contamination is widespread and affects all water source types, including piped supplies. Global burden of disease estimates may have substantially understated the disease burden associated with inadequate water services.
Advances in the healing of flexor tendon injuries
Authors: Sammer Douglas M. , Chung Kevin C. 
Published: May 2014

AbstractThe intrasynovial flexor tendons of the hand are critical for normal hand function. Injury to these tendons can result in absent finger flexion, and a subsequent loss of overall hand function. The surgical techniques used to repair these tendons have improved in the past few decades, as have the postoperative rehabilitation protocols. In spite of these advances, intrasynovial flexor tendon repairs continue to be plagued by postoperative scar formation, which limits tendon gliding and prevents a full functional recovery. This paper describes the current challenges of flexor tendon repair, and evaluates the most recent advances and strategies for achieving an excellent functional outcome.
Rhesus monkeys and baboons develop clotting factor VIII inhibitors in response to porcine endothelial cells or islets
Authors: Stewart John M. , Tarantal Alice F. , Hawthorne Wayne J. , Salvaris Evelyn J. , O'Connell Philip J. , Nottle Mark B. , d'Apice Anthony J. F. , Cowan Peter J. , Kearns‐Jonker Mary 
Published: May 2014

AbstractBackgroundXenotransplantation of porcine organs holds promise of solving the human organ donor shortage. The use of α‐1,3‐galactosyltransferase knockout (GTKO) pig donors mitigates hyperacute rejection, while delayed rejection is currently precipitated by potent immune and hemostatic complications. Previous analysis by our laboratory suggests that clotting factor VIII (FVIII) inhibitors might be elicited by the structurally restricted xenoantibody response which occurs after transplantation of either pig GTKO/hCD55/hCD59/hHT transgenic neonatal islet cell clusters or GTKO endothelial cells.MethodsA recombinant xenoantibody was generated using sequences from baboons demonstrating an active xenoantibody response at day 28 after GTKO/hCD55/hCD59/hHT transgenic pig neonatal islet cell cluster transplantation. Rhesus monkeys were immunized with GTKO pig endothelial cells to stimulate an anti‐non‐Gal xenoantibody response. Serum was collected at days 0 and 7 after immunization. A two‐stage chromogenic assay was used to measure FVIII cofactor activity and identify antibodies which inhibit FVIII function. Molecular modeling and molecular dynamics simulations were used to predict antibody structure and the residues which contribute to antibody‐FVIII interactions. Competition ELISA was used to verify predictions at the domain structural level.ResultsAntibodies that inhibit recombinant human FVIII function are elicited after non‐human primates are transplanted with either GTKO pig neonatal islet cell clusters or endothelial cells. There is an apparent increase in inhibitor titer by 15 Bethesda units (Bu) after transplant, where an increase greater than 5 Bu can indicate pathology in humans. Furthermore, competition ELISA verifies the computer modeled prediction that the recombinant xenoantibody, H66K12, binds the C1 domain of FVIII.ConclusionsThe development of FVIII inhibitors is a novel illustration of the potential impact the humoral immune response can have on coagulative dysfunction in xenotransplantation. However, the contribution of these antibodies to rejection pathology requires further evaluation because “normal” coagulation parameters after successful xenotransplantation are not fully understood.
Recombinant human antithrombin prevents xenogenic activation of hemostasis in a model of pig‐to‐human kidney transplantation
Authors: Ramackers Wolf , Friedrich Lars , Klose Johannes , Vondran Florian , Bergmann Sabine , Schüttler Wolfgang , Johanning Kai , Werwitzke Sonja , Trummer Arne , Bröcker Verena , Klempnauer Jürgen , Winkler Michael , Tiede Andreas 
Published: May 2014

AbstractBackgroundXenogenic activation of hemostasis (XAH) represents a major hurdle for the transplantation of discordant animal organs into humans as it results in thrombotic microangiopathy (TMA). We have previously shown that recombinant human‐activated protein C (rhAPC) mitigates XAH and TMA in an ex vivo model of pig‐to‐human kidney transplantation. However, the use of rhAPC may not be feasible in a perioperative setting due to possible bleeding complications.MethodsHere, we explored the effects of another natural inhibitor of coagulation, human recombinant antithrombin (rhAT), in comparison with rhAPC. Unmodified porcine kidneys (n = 25) were perfused ex vivo with porcine blood, human blood, or human blood supplemented with rhAPC or rhAT. Surrogate parameters of organ survival, markers of XAH (D‐ Dimer, thrombin‐antithrombin complex [TAT], fibrinogen, antithrombin activity, plasminogen), endothelial cell and platelet activation (E‐selectin, P‐selectin), platelet function tests and histological signs of TMA were evaluated.ResultsPerfusion was feasible for > 240 min in all experiments with autologous porcine blood, but limited to 126 ± 78 min with human blood due to increased vascular resistance. Addition of rhAT protected from TMA and allowed for perfusion times > 240 min. In addition, there were less signs of XAH with reduced release of P‐selectin and overexpression of E‐selectin, whereas the progressive loss of platelet function, observed during discordant perfusion, was prevented. The effect of rhAT was dose‐dependent with maximum protection obtained at 3 IU/ml.ConclusionIn conclusion, in this ex vivo model of discordant xenotransplantation, rhAT reduced XAH and prevented TMA in doses that appear feasible for use in clinical or preclinical transplantation settings.
Bipolar fresh osteochondral allograft of the shoulder
Authors: Giannini Sandro , Sebastiani Elisa , Shehu Alba , Baldassarri Matteo , Maraldi Susanna , Vannini Francesca 
Published: May 2014

Purpose: to describe an original technique and preliminary results of bipolar fresh osteochondral allograft implantation for the treatment of end-stage glenohumeral osteoarthritis.
Methods: three patients underwent bipolar fresh osteochondral allograft implantation to the shoulder. Clinical and radiographical evaluations were carried out periodically through to final follow-up.
Results: constant Score increased from 38.3 ± 2.9 preoperatively to 78.7 ± 16.2 at 12 months, 72.3 ±15.3 at 24 months, and 59.3 ± 22.0 at 34 months. Arthritis and partial reabsorption of the implanted surfaces were evident radiographically.
Conclusions: the clinical results obtained in these patients seem to support the applicability of bipolar fresh osteochondral allograft implantation in the shoulder in subjects with severe post-traumatic arthritis and intact rotator cuff. The development of arthritis of the implanted surfaces, while not impacting the clinical result, is a cause of concern.
Level of Evidence: level IV, therapeutic case series.
Neisseria Adhesin A Variation and Revised Nomenclature Scheme
Authors: Bambini Stefania , De Chiara Matteo , Muzzi Alessandro , Mora Marirosa , Lucidarme Jay , Brehony Carina , Borrow Ray , Masignani Vega , Comanducci Maurizio , Maiden Martin C. J. , Rappuoli Rino , Pizza Mariagrazia , Jolley Keith A. 
Published: May 2014

ABSTRACTNeisseriaadhesin A (NadA), involved in the adhesion and invasion ofNeisseria meningitidisinto host tissues, is one of the major components of Bexsero, a novel multicomponent vaccine licensed for protection against meningococcal serogroup B in Europe, Australia, and Canada. NadA has been identified in approximately 30% of clinical isolates and in a much lower proportion of carrier isolates. Three protein variants were originally identified in invasive meningococci and named NadA-1, NadA-2, and NadA-3, whereas most carrier isolates either lacked the gene or harbored a different variant, NadA-4. Further analysis of isolates belonging to the sequence type 213 (ST-213) clonal complex identified NadA-5, which was structurally similar to NadA-4, but more distantly related to NadA-1, -2, and -3. At the time of this writing, more than 89 distinctnadAallele sequences and 43 distinct peptides have been described. Here, we present a revised nomenclature system, taking into account the complete data set, which is compatible with previous classification schemes and is expandable. The main features of this new scheme include (i) the grouping of the previously named NadA-2 and NadA-3 variants into a single NadA-2/3 variant, (ii) the grouping of the previously assigned NadA-4 and NadA-5 variants into a single NadA-4/5 variant, (iii) the introduction of an additional variant (NadA-6), and (iv) the classification of the variants into two main groups, named groups I and II. To facilitate querying of the sequences and submission of new allele sequences, the nucleotide and amino acid sequences are available athttp://pubmlst.org/neisseria/NadA/.
Expansion and Evolution of a Virulent, Extensively Drug-Resistant (Polymyxin B-Resistant), QnrS1-, CTX-M-2-, and KPC-2-Producing Klebsiella pneumoniae ST11 International High-Risk Clone
Authors: Andrade Leonardo Neves , Vitali Lúcia , Gaspar Gilberto Gambero , Bellissimo-Rodrigues Fernando , Martinez Roberto , Darini Ana Lúcia Costa 
Published: May 2014

ABSTRACT

In this study, we report the early expansion, evolution, and characterization of a multiresistant
Klebsiella pneumoniae
clone that was isolated with increasing frequency from inpatients in a tertiary-care university hospital in Brazil. Seven carbapenem- and quinolone-resistant and polymyxin B-susceptible or -resistant
K. pneumoniae
isolates isolated between December 2012 and February 2013 were investigated. Beta-lactamase- and plasmid-mediated quinolone resistance (PMQR)-encoding genes and the genetic environment were investigated using PCR, sequencing, and restriction fragment length polymorphism (RFLP). Clonal relatedness was established using XbaI–pulsed-field gel electrophoresis (PFGE), multilocus sequence typing (MLST), and phylogenetic group characterization. Plasmid analyses included PCR-based replicon typing (PBRT) and hybridization of the
S1
-PFGE product, plasmid MLST, and conjugation experiments. Virulence potential was assessed by PCR by searching for 10 virulence factor-encoding genes (
ureA
,
fimH
,
kfuBC
,
uge
,
wabG
,
magA
,
mrkD
,
allS
,
rmpA
, and
cf29a
) and by phenotypic tests to analyze the hypermucoviscous phenotype. The genetic context of a multidrug-resistant and extensively drug-resistant
K. pneumoniae
ST11-KpI clone harboring IncFIIk-Tn
4401a-bla
KPC-2
,
qnrS1
, and
bla
CTX-M-2
was found. Moreover, three isolates displayed high resistance to polymyxin B (MICs = 32, 32, and 128 mg/liter) as well as mucous and hypermucoviscous phenotypes. These bacteria also harbored
ureA
,
fimH
,
uge
,
wabG
, and
mrkD
, which code for virulence factors associated with binding, biofilm formation, and the ability to colonize and escape from phagocytosis. Our study describes the association of important coresistance and virulence factors in the
K. pneumoniae
ST11 international high-risk clone, which makes this pathogen successful at infections and points to the quick expansion and evolution of this multiresistant and virulent clone, leading to a pandrug-resistant phenotype and persistent bacteria in a Brazilian hospital.
Utility of
rpoB
Gene Sequencing for Identification of Nontuberculous Mycobacteria in the Netherlands
Authors: de Zwaan Rina , van Ingen Jakko , van Soolingen Dick 
Published: May 2014

ABSTRACT

In the Netherlands, clinical isolation of nontuberculous mycobacteria (NTM) has increased over the past decade. Proper identification of isolates is important, as NTM species differ strongly in clinical relevance. Most of the currently applied identification methods cannot distinguish between all different
Mycobacterium
species and complexes within species.
rpoB
gene sequencing exhibits a promising level of discrimination among rapidly and slowly growing mycobacteria, including the
Mycobacterium avium
complex. In this study, we prospectively compared
rpoB
gene sequencing with our routine algorithm of reverse line blot identification combined with partial 16S rRNA gene sequencing of 455 NTM isolates.
rpoB
gene sequencing identified 403 isolates to species level as 45 different known species and identified 44 isolates to complex level, and eight isolates remained unidentifiable to species level. In contrast, our reference reverse line blot assay with adjunctive 16S rRNA gene sequencing identified 390 isolates to species level (30 distinct species) and identified 56 isolates to complex level, and nine isolates remained unidentified. The higher discriminatory power of
rpoB
gene sequencing results largely from the distinction of separate species within complexes and subspecies. Also,
Mycobacterium gordonae
,
Mycobacterium kansasii
, and
Mycobacterium interjectum
were separated into multiple groupings with relatively low sequence similarity (98 to 94%), suggesting that these are complexes of closely related species. We conclude that
rpoB
gene sequencing is a more discriminative identification technique than the combination of reverse line blot and 16S rRNA gene sequencing and could introduce a major improvement in clinical care of NTM disease and the research on the epidemiology and clinical relevance of NTM.
Evaluation of Three Rapid Diagnostic Methods for Direct Identification of Microorganisms in Positive Blood Cultures
Authors: Martinez Raquel M. , Bauerle Elizabeth R. , Fang Ferric C. , Butler-Wu Susan M. 
Published: May 2014

ABSTRACT

The identification of organisms from positive blood cultures generally takes several days. However, recently developed rapid diagnostic methods offer the potential for organism identification within only a few hours of blood culture positivity. In this study, we evaluated the performance of three commercial methods to rapidly identify organisms directly from positive blood cultures:
Quick
FISH (AdvanDx, Wolburn, MA), Verigene Gram-Positive Blood Culture (BC-GP; Nanosphere, Northbrook, IL), and matrix-assisted laser desorption ionization–time of flight mass spectrometry (MALDI-TOF MS) with Sepsityper processing (Bruker Daltonics, Billerica, MA). A total of 159 blood cultures (VersaTREK Trek Diagnostic Systems, Cleveland, OH) positive for Gram-positive and Gram-negative bacteria as well as yeast were analyzed with
Quick
FISH and MALDI-TOF MS. In all, 102 blood cultures were analyzed using the BC-GP assay. For monomicrobial cultures, we observed 98.0% concordance with routine methods for both
Quick
FISH (143/146) and the BC-GP assay (93/95). MALDI-TOF MS demonstrated 80.1% (117/146) and 87.7% (128/146) concordance with routine methods to the genus and species levels, respectively. None of the methods tested were capable of consistently identifying polymicrobial cultures in their entirety or reliably differentiating
Streptococcus pneumoniae
from viridans streptococci. Nevertheless, the methods evaluated in this study are convenient and accurate for the most commonly encountered pathogens and have the potential to dramatically reduce turnaround time for the provision of results to the treating physician.