bluuebunny/crossref_metadata_2025_split huggingface dataset:
Human bone marrow processing using a new continuous‐flow cell separation device
Authors: Guttridge Martin G. , Bailey Christopher , Sidders Christopher , Nichols Jennifer , Bromham Jane , Watt Suzanne M. 
Published: December 2015

BACKGROUNDProcessing of bone marrow (BM) is often required to remove incompatible red blood cells (RBCs) or to reduce the volume before transplantation or cryopreservation. We have evaluated the Spectra Optia apheresis system to determine its effectiveness in volume reduction and RBC depletion of human BM before transplantation.STUDY DESIGN AND METHODSBM from 30 donations (28 allogeneic and two autologous) were processed using the Spectra Optia over a 12‐month period. The mean BM collection volume was 1094 ± 337 mL and RBC volume was 374 ± 148 mL. Processing using the Spectra Optia was as described by the manufacturer.RESULTSVolume reduction achieved was 93.0 ± 1.2%; RBC depletion was 98.8 ± 0.4%; and mononuclear, CD34+, and CD3+ cell recoveries were 79.12 ± 14.03, 88.36 ± 13.76, and 79.84 ± 16.27%, respectively. In total 26 of 28 processed allografts were transplanted; 24 achieved neutrophil engraftment in 20.7 ± 5.9 days and 18 achieved platelet engraftment in 19.6 ± 8.9 days. Time in transit significantly affected the Spectra Optia's ability to recover mononuclear, CD34+, and CD3+ cells, and the overall age of the BM at the time of processing significantly affected the recovery of mononuclear and CD3+ cells, but not CD34+ cells. Time in storage at 2 to 6°C had no adverse effect on processing.CONCLUSIONThis study demonstrates that the Spectra Optia can effectively volume reduce and RBC deplete human BM before transplantation. Time in transit should be as short as possible but may be extended up to 24 hours if the donation is refrigerated during transit.
Metabolomics in transfusion medicine
Authors: Nemkov Travis , Hansen Kirk C. , Dumont Larry J. , D'Alessandro Angelo 
Published: December 2015

Biochemical investigations on the regulatory mechanisms of red blood cell (RBC) and platelet (PLT) metabolism have fostered a century of advances in the field of transfusion medicine. Owing to these advances, storage of RBCs and PLT concentrates has become a lifesaving practice in clinical and military settings. There, however, remains room for improvement, especially with regard to the introduction of novel storage and/or rejuvenation solutions, alternative cell processing strategies (e.g., pathogen inactivation technologies), and quality testing (e.g., evaluation of novel containers with alternative plasticizers). Recent advancements in mass spectrometry–based metabolomics and systems biology, the bioinformatics integration of omics data, promise to speed up the design and testing of innovative storage strategies developed to improve the quality, safety, and effectiveness of blood products.Here we review the currently available metabolomics technologies and briefly describe the routine workflow for transfusion medicine–relevant studies. The goal is to provide transfusion medicine experts with adequate tools to navigate through the otherwise overwhelming amount of metabolomics data burgeoning in the field during the past few years.Descriptive metabolomics data have represented the first step omics researchers have taken into the field of transfusion medicine. However, to up the ante, clinical and omics experts will need to merge their expertise to investigate correlative and mechanistic relationships among metabolic variables and transfusion‐relevant variables, such as 24‐hour in vivo recovery for transfused RBCs. Integration with systems biology models will potentially allow for in silico prediction of metabolic phenotypes, thus streamlining the design and testing of alternative storage strategies and/or solutions.
Endothelin regulates intermittent hypoxia‐induced lipolytic remodelling of adipose tissue and phosphorylation of hormone‐sensitive lipase
Authors: Briançon‐Marjollet Anne , Monneret Denis , Henri Marion , Hazane‐Puch Florence , Pepin Jean‐Louis , Faure Patrice , Godin‐Ribuot Diane 
Published: December 2015

Key points
Endothelin‐1 (ET‐1) is upregulated upon intermittent hypoxia and has lipolytic effects.
Intermittent hypoxia induces adipose tissue lipolysis that may be mediated by ET‐1.
In the present study, we show that ET‐1 is involved in adipose tissue remodelling induced by intermittent hypoxia and that phosphorylation of hormone‐sensitive lipase could be one mechanism mediating this effect.
We also show that ET‐1 upregulates its own and its type A endothelin receptor expression, possibly leading to an autoactivatory loop.
These results help us better understand the mechanisms of dyslipidaemia in disorders associated with intermittent hypoxia, such as obstructive sleep apnoea.
AbstractObstructive sleep apnoea syndrome is characterized by repetitive episodes of upper airway collapse during sleep resulting in chronic intermittent hypoxia (IH). Obstructive sleep apnoea syndrome, through IH, promotes cardiovascular and metabolic disorders. Endothelin‐1 (ET‐1) secretion is upregulated by IH, and is able to modulate adipocyte metabolism. Therefore, the present study aimed to characterize the role of ET‐1 in the metabolic consequences of IH on adipose tissue in vivo and in vitro. Wistar rats were submitted to 14 days of IH‐cycles (30 s of 21% FiO2 and 30 s of 5% FiO2; 8 h day−1) or normoxia (air–air cycles) and were treated or not with bosentan, a dual type A and B endothelin receptor (ETA‐R and ETB‐R) antagonist. Bosentan treatment decreased plasma free fatty acid and triglyceride levels, and inhibited IH‐induced lipolysis in adipose tissue. Moreover, IH induced a 2‐fold increase in ET‐1 transcription and ETA‐R expression in adipose tissue that was reversed by bosentan. In 3T3‐L1 adipocytes, ET‐1 upregulated its own and its ETA‐R transcription and this effect was abolished by bosentan. Moreover, ET‐1 induced glycerol release and inhibited insulin‐induced glucose uptake. Bosentan and BQ123 inhibited these effects. Bosentan also reversed the ET‐1‐induced phosphorylation of hormone‐sensitive lipase (HSL) on Ser660. Finally, ET‐1‐induced lipolysis and HSL phosphorylation were also observed under hypoxia. Altogether, these data suggest that ET‐1 is involved in IH‐induced lipolysis in Wistar rats, and that upregulation of ET‐1 production and ETA‐R expression by ET‐1 itself under IH could amplify its effects. Moreover, ET‐1‐induced lipolysis could be mediated through ETA‐R and activation of HSL by Ser660 phosphorylation.
Increasing taurine intake and taurine synthesis improves skeletal muscle function in the mdx mouse model for Duchenne muscular dystrophy
Authors: Terrill Jessica R. , Pinniger Gavin J. , Graves Jamie A. , Grounds Miranda D. , Arthur Peter G. 
Published: December 2015

Key points
Duchenne muscular dystrophy (DMD) is a fatal muscle wasting disease associated with increased inflammation, oxidative stress and myofibre necrosis.
Cysteine precursor antioxidants such as N‐acetyl cysteine (NAC) and l‐2‐oxothiazolidine‐4‐carboxylate (OTC) reduce dystropathology in the mdx mouse model for DMD, and we propose this is via increased synthesis of the amino acid taurine.
We compared the capacity of OTC and taurine treatment to increase taurine content of mdx muscle, as well as effects on in vivo and ex vivo muscle function, inflammation and oxidative stress.
Both treatments increased taurine in muscles, and improved many aspects of muscle function and reduced inflammation. Taurine treatment also reduced protein thiol oxidation and was overall more effective, as OTC treatment reduced body and muscle weight, suggesting some adverse effects of this drug.
These data suggest that increasing dietary taurine is a better candidate for a therapeutic intervention for DMD.
AbstractDuchenne muscular dystrophy (DMD) is a fatal muscle wasting disease for which there is no widely available cure. Whilst the mechanism of loss of muscle function in DMD and the mdx mouse model are not fully understood, disruptions in intracellular calcium homeostasis, inflammation and oxidative stress are implicated. We have shown that protein thiol oxidation is increased in mdx muscle, and that the indirect thiol antioxidant l‐2‐oxothiazolidine‐4‐carboxylate (OTC), which increases cysteine availability, decreases pathology and increases in vivo strength. We propose that the protective effects of OTC are a consequence of conversion of cysteine to taurine, which has itself been shown to be beneficial to mdx pathology. This study compares the efficacy of taurine with OTC in decreasing dystropathology in mdx mice by measuring in vivo and ex vivo contractile function and measurements of inflammation and protein thiol oxidation. Increasing the taurine content of mdx muscle improved both in vivo and ex vivo muscle strength and function, potentially via anti‐inflammatory and antioxidant effects of taurine. OTC treatment increased taurine synthesis in the liver and taurine content of mdx muscle, improved muscle function and decreased inflammation. However, OTC was less effective than taurine treatment, with OTC also decreasing body and EDL muscle weights, suggesting that OTC had some detrimental effects. These data support continued research into the use of taurine as a therapeutic intervention for DMD, and suggest that increasing dietary taurine is the better strategy for increasing taurine content and decreasing severity of dystropathology.
Microglia modulate brainstem serotonergic expression following neonatal sustained hypoxia exposure: implications for sudden infant death syndrome
Authors: MacFarlane P. M. , Mayer C. A. , Litvin D. G. 
Published: December 2015

Key points
Neonatal sustained hypoxia exposure modifies brainstem microglia and serotonin expression.
The altered brainstem neurochemistry is associated with impaired ventilatory responses to acute hypoxia and mortality.
The deleterious effects of sustained hypoxia exposure can be prevented by an inhibitor of activated microglia.
These observations demonstrate a potential cause of the brainstem serotonin abnormalities thought to be involved in sudden infant death syndrome.
AbstractWe showed previously that the end of the second postnatal week (days P11–15) represents a period of development during which the respiratory neural control system exhibits a heightened vulnerability to sustained hypoxia (SH, 11% O2, 5 days) exposure. In the current study, we investigated whether the vulnerability to SH during the same developmental time period is associated with changes in brainstem serotonin (5‐HT) expression and whether it can be prevented by the microglia inhibitor minocycline. Using whole‐body plethysmography, SH attenuated the acute (5 min) hypoxic ventilatory response (HVR) and caused a high incidence of mortality compared to normoxia rats. SH also increased microglia cell numbers and decreased 5‐HT immunoreactivity in the nucleus of the solitary tract (nTS) and dorsal motor nucleus of the vagus (DMNV). The attenuated HVR, mortality, and changes in nTS and DMNV immunoreactivity was prevented by minocycline (25 mg kg−1/2 days during SH). These data demonstrate that the 5‐HT abnormalities in distinct respiratory neural control regions can be initiated by prolonged hypoxia exposure and may be modulated by microglia activity. These observations share several commonalities with the risk factors thought to underlie the aetiology of sudden infant death syndrome, including: (1) a vulnerable neonate; (2) a critical period of development; (3) evidence of hypoxia; (4) brainstem gliosis (particularly the nTS and DMNV); and (5) 5‐HT abnormalities.
An Interspecies Signaling System Mediated by Fusaric Acid Has Parallel Effects on Antifungal Metabolite Production by Pseudomonas protegens Strain Pf-5 and Antibiosis of Fusarium spp
Authors: Quecine Maria Carolina , Kidarsa Teresa A. , Goebel Neal C. , Shaffer Brenda T. , Henkels Marcella D. , Zabriskie T. Mark , Loper Joyce E. 
Published: December 2015

ABSTRACT

Pseudomonas protegens
strain Pf-5 is a rhizosphere bacterium that suppresses soilborne plant diseases and produces at least seven different secondary metabolites with antifungal properties. We derived mutants of Pf-5 with single and multiple mutations in biosynthesis genes for seven antifungal metabolites: 2,4-diacetylphoroglucinol (DAPG), pyrrolnitrin, pyoluteorin, hydrogen cyanide, rhizoxin, orfamide A, and toxoflavin. These mutants were tested for inhibition of the pathogens
Fusarium verticillioides
and
Fusarium oxysporum
f. sp.
pisi
. Rhizoxin, pyrrolnitrin, and DAPG were found to be primarily responsible for fungal antagonism by Pf-5. Previously, other workers showed that the mycotoxin fusaric acid, which is produced by many
Fusarium
species, including
F. verticillioides
, inhibited the production of DAPG by
Pseudomonas
spp. In this study, amendment of culture media with fusaric acid decreased DAPG production, increased pyoluteorin production, and had no consistent influence on pyrrolnitrin or orfamide A production by Pf-5. Fusaric acid also altered the transcription of biosynthetic genes, indicating that the mycotoxin influenced antibiotic production by Pf-5 at the transcriptional level. Addition of fusaric acid to the culture medium reduced antibiosis of
F. verticillioides
by Pf-5 and derivative strains that produce DAPG but had no effect on antibiosis by Pf-5 derivatives that suppressed
F. verticillioides
due to pyrrolnitrin or rhizoxin production. Our results demonstrated the importance of three compounds, rhizoxin, pyrrolnitrin, and DAPG, in suppression of
Fusarium
spp. by Pf-5 and confirmed that an interspecies signaling system mediated by fusaric acid had parallel effects on antifungal metabolite production and antibiosis by the bacterial biological control organism.
Prevalence and Genetic Diversity of Human Sapoviruses in Shellfish from Commercial Production Areas in Galicia, Spain
Authors: Varela Miguel F. , Polo David , Romalde Jesús L. 
Published: December 2015

ABSTRACTThe prevalence of human forms ofSapovirus, an emerging pathogen of human gastroenteritis, was investigated in an 18-month survey from class B mollusc-harvesting areas in two Galician rias (northwest Spain). The detection and quantification ofSapoviruswas performed by reverse transcription-real-time PCR, according to the recently developed standard method ISO/TS 15216-1:2013, and genotyping by reverse transcription-nested PCR. The bivalve species studied were wild and cultured mussels (Mytilus galloprovincialis), clams (Venerupis philippinarumandVenerupis decussata), and cockles (Cerastoderma edule).Sapoviruswas detected in 30 out of 168 samples (17.9%), with cockles being the species with the highest prevalence of positives (28.1%), followed by clams (22.6%), wild mussels (14.3%), and cultured mussels (12.9%). The estuary in the south of the region demonstrated a higher percentage of positive samples (21.8%) than the one in the north (14.4%). Viral contamination levels for the positive samples ranged between 1.9 × 103and 1.4 × 105RNA copies/g of digestive tissue. ThirteenSapovirussequences could be obtained based on partial capsid gene sequence and were classified into four genotypes: GI.1 (2 samples), GI.2 (8 samples), GIV.1 (2 samples), and GV.1 (1 sample).
A 2-Hydroxypyridine Catabolism Pathway in Rhodococcus rhodochrous Strain PY11
Authors: Vaitekūnas Justas , Gasparavičiūtė Renata , Rutkienė Rasa , Tauraitė Daiva , Meškys Rolandas 
Published: December 2015

ABSTRACT

Rhodococcus rhodochrous
PY11 (DSM 101666) is able to use 2-hydroxypyridine as a sole source of carbon and energy. By investigating a gene cluster (
hpo
) from this bacterium, we were able to reconstruct the catabolic pathway of 2-hydroxypyridine degradation. Here, we report that in
Rhodococcus rhodochrous
PY11, the initial hydroxylation of 2-hydroxypyridine is catalyzed by a four-component dioxygenase (HpoBCDF). A product of the dioxygenase reaction (3,6-dihydroxy-1,2,3,6-tetrahydropyridin-2-one) is further oxidized by HpoE to 2,3,6-trihydroxypyridine, which spontaneously forms a blue pigment. In addition, we show that the subsequent 2,3,6-trihydroxypyridine ring opening is catalyzed by the hypothetical cyclase HpoH. The final products of 2-hydroxypyridine degradation in
Rhodococcus rhodochrous
PY11 are ammonium ion and α-ketoglutarate.
Lactobacillus rhamnosus GR-1 Ameliorates Escherichia coli-Induced Inflammation and Cell Damage via Attenuation of ASC-Independent NLRP3 Inflammasome Activation
Authors: Wu Qiong , Liu Ming-Chao , Yang Jun , Wang Jiu-Feng , Zhu Yao-Hong 
Published: December 2015

ABSTRACT

Escherichia coli
is a major environmental pathogen causing bovine mastitis, which leads to mammary tissue damage and cell death. We explored the effects of the probiotic
Lactobacillus rhamnosus
GR-1 on ameliorating
E. coli
-induced inflammation and cell damage in primary bovine mammary epithelial cells (BMECs). Increased Toll-like receptor 4 (TLR4), NOD1, and NOD2 mRNA expression was observed following
E. coli
challenge, but this increase was attenuated by
L. rhamnosus
GR-1 pretreatment. Immunofluorescence and Western blot analyses revealed that
L. rhamnosus
GR-1 pretreatment decreased the
E. coli
-induced increases in the expression of the NOD-like receptor family member pyrin domain-containing protein 3 (NLRP3) and the serine protease caspase 1. However, expression of the adaptor protein apoptosis-associated speck-like protein (ASC, encoded by the
Pycard
gene) was decreased during
E. coli
infection, even with
L. rhamnosus
GR-1 pretreatment. Pretreatment with
L. rhamnosus
GR-1 counteracted the
E. coli
-induced increases in interleukin-1β (IL-1β), -6, -8, and -18 and tumor necrosis factor alpha mRNA expression but upregulated IL-10 mRNA expression. Our data indicate that
L. rhamnosus
GR-1 reduces the adhesion of
E. coli
to BMECs, subsequently ameliorating
E. coli
-induced disruption of cellular morphology and ultrastructure and limiting detrimental inflammatory responses, partly via promoting TLR2 and NOD1 synergism and attenuating ASC-independent NLRP3 inflammasome activation. Although the residual pathogenic activity of
L. rhamnosus
, the dosage regimen, and the means of probiotic supplementation in cattle remain undefined, our data enhance our understanding of the mechanism of action of this candidate probiotic, allowing for development of specific probiotic-based therapies and strategies for preventing pathogenic infection of the bovine mammary gland.
A NewN-Acyl Homoserine Lactone Synthase in an Uncultured Symbiont of the Red Sea Sponge Theonella swinhoei
Authors: Britstein Maya , Devescovi Giulia , Handley Kim M. , Malik Assaf , Haber Markus , Saurav Kumar , Teta Roberta , Costantino Valeria , Burgsdorf Ilia , Gilbert Jack A. , Sher Noa , Venturi Vittorio , Steindler Laura 
Published: December 2015

ABSTRACTSponges harbor a remarkable diversity of microbial symbionts in which signal molecules can accumulate and enable cell-cell communication, such as quorum sensing (QS). Bacteria capable of QS were isolated from marine sponges; however, an extremely small fraction of the sponge microbiome is amenable to cultivation. We took advantage of community genome assembly and binning to investigate the uncultured majority of sponge symbionts. We identified a completeN-acyl-homoserine lactone (AHL)-QS system (designated TswIR) and seven partialluxIhomologues in the microbiome ofTheonella swinhoei. The TswIR system was novel and shown to be associated with an alphaproteobacterium of the orderRhodobacterales, here termedRhodobacteralesbacterium TS309. ThetswIgene, when expressed inEscherichia coli, produced three AHLs, two of which were also identified in aT. swinhoeisponge extract. The taxonomic affiliation of the 16S rRNA ofRhodobacteralesbacterium TS309 to a sponge-coral specific clade, its enrichment in sponge versus seawater and marine sediment samples, and the presence of sponge-specific features, such as ankyrin-like domains and tetratricopeptide repeats, indicate a likely symbiotic nature of this bacterium.
Alginate Biosynthesis Factories in Pseudomonas fluorescens: Localization and Correlation with Alginate Production Level
Authors: Maleki Susan , Almaas Eivind , Zotchev Sergey , Valla Svein , Ertesvåg Helga 
Published: December 2015

ABSTRACT

Pseudomonas fluorescens
is able to produce the medically and industrially important exopolysaccharide alginate. The proteins involved in alginate biosynthesis and secretion form a multiprotein complex spanning the inner and outer membranes. In the present study, we developed a method by which the porin AlgE was detected by immunogold labeling and transmission electron microscopy. Localization of the AlgE protein was found to depend on the presence of other proteins in the multiprotein complex. No correlation was found between the number of alginate factories and the alginate production level, nor were the numbers of these factories affected in an
algC
mutant that is unable to produce the precursor needed for alginate biosynthesis. Precursor availability and growth phase thus seem to be the main determinants for the alginate production rate in our strain. Clustering analysis demonstrated that the alginate multiprotein complexes were not distributed randomly over the entire outer cell membrane surface.
A Fivefold Parallelized Biosynthetic Process Secures Chlorination of Armillaria mellea (Honey Mushroom) Toxins
Authors: Wick Jonas , Heine Daniel , Lackner Gerald , Misiek Mathias , Tauber James , Jagusch Hans , Hertweck Christian , Hoffmeister Dirk 
Published: December 2015

ABSTRACT

The basidiomycetous tree pathogen
Armillaria mellea
(honey mushroom) produces a large variety of structurally related antibiotically active and phytotoxic natural products, referred to as the melleolides. During their biosynthesis, some members of the melleolide family of compounds undergo monochlorination of the aromatic moiety, whose biochemical and genetic basis was not known previously. This first study on basidiomycete halogenases presents the biochemical
in vitro
characterization of five flavin-dependent
A. mellea
enzymes (ArmH1 to ArmH5) that were heterologously produced in
Escherichia coli
. We demonstrate that all five enzymes transfer a single chlorine atom to the melleolide backbone. A 5-fold, secured biosynthetic step during natural product assembly is unprecedented. Typically, flavin-dependent halogenases are categorized into enzymes acting on free compounds as opposed to those requiring a carrier-protein-bound acceptor substrate. The enzymes characterized in this study clearly turned over free substrates. Phylogenetic clades of halogenases suggest that all fungal enzymes share an ancestor and reflect a clear divergence between ascomycetes and basidiomycetes.