bluuebunny/crossref_metadata_2025_split huggingface dataset:
The role of the metastasis suppressor gene KAI1 in melanoma angiogenesis
Authors: Tang Yun , Bhandaru Madhuri , Cheng Yabin , Lu Jing , Li Gang , Ong Christopher J. 
Published: July 2015

SummaryThe tetraspan protein KAI1 (CD82) has been previously shown to have important roles in cell migration, invasion, and melanoma prognosis. In this study, we investigated the role of KAI1 regarding melanoma angiogenesis. KAI1 overexpression strongly suppressed the growth of the human umbilical vein endothelial cells and their tubular structure formation in vitro. Also, KAI1 was able to inhibit both interleukin‐6 (IL‐6) and VEGF at mRNA and protein levels. Using nude mice in the in vivo study, we showed that KAI1, through the regulation of ING4, inhibited blood vessel formation in matrigel plugs along with the downregulation of IL‐6 and VEGF, and the recruitment of CD31‐positive cells. Finally, we found that KAI1 was able to suppress the activity of a serine/threonine kinase Akt by suppressing Akt phosphorylation (Ser473). Taken together, our results suggested that KAI1 was able to suppress melanoma angiogenesis by downregulating IL‐6 and VEGF expression, and the restoration of KAI1 functionality offered a new approach in human melanoma treatment.
Haemovigilance study at a tertiary care hospital in the north‐east of India
Authors: Somagari D. R. , Sriram C. S. , Rachamalla C. K. V. , Dutta U. C. , Chaliha T. , Lahkar M. , Bezbaruah B. K. 
Published: July 2015

Background and AimBlood transfusion is a frequent and integral part of critical patient care. Even though life‐saving, it could be occasionally risky and result in a spectrum of adverse events. Haemovigilance programme is a standard set‐up to monitor the entire transfusion chain, evaluate and analyse the data to improve patient's safety. The haemovigilance programme has been established in India very recently. We report the implemented haemovigilance programme at a tertiary care hospital in the north‐east of India.Methods and MaterialsThis study was prospective and observational. It was carried out for a period of 9 months, from September 2013 to May 2014, in the State of the Art Model Blood Bank, Gauhati Medical College and Hospital, Guwahati, Assam, India. Transfusion reaction reporting forms, which are supplied by the Indian Pharmacopoeia Commission, were utilized to collect the data.ResultsA total of 51 000 blood components were transfused and 106 adverse events reported during the period of the study. The major stakes of adverse events were febrile non‐haemolytic transfusion reactions (50%), pruritus (17·92%) and rigour/chills (11·32%). All the adverse events were found to be Grade 1 (non‐severe) type, and there were no severe or life‐threatening adverse events.
Is there something fishy about the regulation of the ryanodine receptor in the fish heart?
Authors: Shiels Holly A. , Sitsapesan Rebecca 
Published: July 2015

New Findings

What is the topic of this review?
Excitation–contraction coupling in fish hearts is maintained over a range of temperatures that would be cardioplegic to most mammals. Here, we review what is known about the fish cardiac ryanodine receptor, and consider how it may be regulated in a different manner from the mammalian cardiac isoforms of this channel.

What advances does it highlight?
We highlight how a better understanding of the basic gating and conducting properties of fish cardiac ryanodine receptors could provide considerable insight into mechanisms underlying sarcoplasmic reticulum calcium release in fish hearts and the role of the sarcoplasmic reticulum in the evolution of the heart.

The fish cardiac sarcoplasmic reticulum (SR) holds large quantities of Ca2+, but calcium‐induced calcium release (CICR) is weak in these myocytes, and contraction and relaxation are largely determined by transsarcolemmal Ca2+ flux. Many fish species live in a cold and seasonally variable thermal habitat, which could provide challenges to regulation of excitation–contraction coupling. Here, we focus on the cardiac SR Ca2+‐release channel (RyR2) in fish and ask whether it may be regulated in a different manner from the mammalian RyR2. We review data indicating that fish cardiac RyR are present at lower density, are more spatially separated within the SR membrane and are less responsive to cytosolic Ca2+ than mammalian RyR2 channels. All of these features would contribute to the weak CICR evident from functional studies. We also consider how CICR can be enhanced in fish myocytes following β‐adrenergic stimulation and application of low levels of caffeine, and how acute and chronic temperature change may affect the gating properties of fish RyR2s. It is clear that a lack of insight into the fundamental gating and conductance properties of fish RyR2 channels is hindering our understanding of the role of the SR in fish cardiac excitation–contraction coupling. We conclude by reflecting on how studies that probe the biophysical properties of fish RyR2 channel gating in response to various ligands and temperatures would be very instructive for our understanding of the role of the SR in the evolution of the heart.
A Local-to-Global Dimensional Error Calculation Framework for the Riveted Assembly Using Finite-Element Analysis
Authors: Ni Jun , Tang Wencheng , Xing Yan , Ben Kecun , Li Ming 
Published: July 2015

Abstract
Mechanical structures of large-scale antennas are sheet metals connected by thousands of rivets. The antenna dimensional error after riveting often violates the limit allowed. The prediction of the global dimensional error induced by many rivet connections requires a rapid and accurate assembly deformation calculation method. Main process parameters of these local rivet connections are the local connection dimension, material property, local clamp position, rivet upsetting direction, and the hammer time-to-displacement impact, except for the riveting sequence. We neglect the process parameter uncertainties and consider that the main riveting parameters equate to a dynamic finite-element (FE) model of single rivet connection. The dynamic FE analysis result yields an inherent strain database for the riveted local parts. Then, we propose an iterative loop of static FE analyses for the global structure taking the inherent strain database and possible former static FE analysis result as the boundary conditions. The loop forms a local-to-global framework. Two examples are involved through the framework representation and realistic application. Framework advantages include: (1) a good balance between the cost and precision of dimensional error calculation; (2) the sequence simulation of all the riveting operations; and (3) supporting the further assembly process optimization to reduce the global dimensional error of the assembly with thousands of rivets.
Baseline Polymorphisms and Emergence of Drug Resistance in the NS3/4A Protease of Hepatitis C Virus Genotype 1 following Treatment with Faldaprevir and Pegylated Interferon Alpha 2a/Ribavirin in Phase 2 and Phase 3 Studies
Authors: Berger K. L. , Scherer J. , Ranga M. , Sha N. , Stern J. O. , Quinson A.-M. , Kukolj G. 
Published: July 2015

ABSTRACT

Analysis of data pooled from multiple phase 2 (SILEN-C1 to 3) and phase 3 studies (STARTVerso1 to 4) of the hepatitis C virus (HCV) nonstructural protein 3/4A (NS3/4A) protease inhibitor faldaprevir plus pegylated interferon alpha/ribavirin (PR) provides a comprehensive evaluation of baseline and treatment-emergent NS3/4A amino acid variants among HCV genotype-1 (GT-1)-infected patients. Pooled analyses of GT-1a and GT-1b NS3 population-based pretreatment sequences (
n
= 3,124) showed that faldaprevir resistance-associated variants (RAVs) at NS3 R155 and D168 were rare (<1%). No single, noncanonical NS3 protease or NS4A cofactor baseline polymorphism was associated with a reduced sustained virologic response (SVR) to faldaprevir plus PR, including Q80K. The GT-1b NS3 helicase polymorphism T344I was associated with reduced SVR to faldaprevir plus PR (
P
< 0.0001) but was not faldaprevir specific, as reduced SVR was also observed with placebo plus PR. Among patients who did not achieve SVR and had available NS3 population sequences (
n
= 507 GT-1a;
n
= 349 GT-1b), 94% of GT-1a and 83% of GT-1b encoded faldaprevir treatment-emergent RAVs. The predominant GT-1a RAV was R155K (88%), whereas GT-1b encoded D168 substitutions (78%) in which D168V was predominant (67%). The novel GT-1b NS3 S61L substitution emerged in 7% of virologic failures as a covariant with D168V, most often among the faldaprevir breakthroughs; S61L in combination with D168V had a minimal impact on faldaprevir susceptibility compared with that for D168V alone (1.5-fold difference
in vitro
). The median time to loss of D168 RAVs among GT-1b-infected patients who did not have a sustained virologic response at 12 weeks posttreatment (non-SVR12) after virologic failure was 5 months, which was shorter than the 14 months for R155 RAVs among GT-1a-infected non-SVR12 patients, suggesting that D168V is less fit than R155K in the absence of faldaprevir selective pressure.
In Vitro
Activity of AZD0914, a Novel Bacterial DNA Gyrase/Topoisomerase IV Inhibitor, against Clinically Relevant Gram-Positive and Fastidious Gram-Negative Pathogens
Authors: Biedenbach Douglas J. , Huband Michael D. , Hackel Meredith , de Jonge Boudewijn L. M. , Sahm Daniel F. , Bradford Patricia A. 
Published: July 2015

ABSTRACT

AZD0914, a new spiropyrimidinetrione bacterial DNA gyrase inhibitor with a novel mode of inhibition, has activity against bacterial species commonly cultured from patient infection specimens, including fluoroquinolone-resistant isolates. This study assessed the
in vitro
activity of AZD0914 against key Gram-positive and fastidious Gram-negative clinical isolates collected globally in 2013. AZD0914 demonstrated potent activity, with MIC
90
s for AZD0914 of 0.25 mg/liter against
Staphylococcus aureus
(
n
= 11,680), coagulase-negative staphylococci (
n
= 1,923), streptococci (
n
= 4,380), and
Moraxella catarrhalis
(
n
= 145), 0.5 mg/liter against
Staphylococcus lugdunensis
(
n
= 120) and
Haemophilus influenzae
(
n
= 352), 1 mg/liter against
Enterococcus faecalis
(
n
= 1,241), and 2 mg/liter against
Haemophilus parainfluenzae
(
n
= 70). The activity against
Enterococcus faecium
was more limited (MIC
90
, 8 mg/liter). The spectrum and potency of AZD0914 included fluoroquinolone-resistant isolates in each species group, including methicillin-resistant staphylococci, penicillin-resistant streptococci, vancomycin-resistant enterococci, β-lactamase-producing
Haemophilus
spp., and
M. catarrhalis
. Based on these
in vitro
findings, AZD0914 warrants further investigation for its utility against a variety of Gram-positive and fastidious Gram-negative bacterial species.
Genomic and Functional Portrait of a Highly Virulent, CTX-M-15-Producing
H
30-Rx Subclone of Escherichia coli Sequence Type 131
Authors: Ranjan Amit , Shaik Sabiha , Hussain Arif , Nandanwar Nishant , Semmler Torsten , Jadhav Savita , Wieler Lothar H. , Ahmed Niyaz 
Published: July 2015

ABSTRACT

Escherichia coli
sequence type 131 (ST131) is a pandemic clone associated with multidrug-resistant, extraintestinal infections, attributable to the presence of the CTX-M-15 extended-spectrum β-lactamase gene and mutations entailing fluoroquinolone resistance. Studies on subclones within
E. coli
ST131 are critically required for targeting and implementation of successful control efforts. Our study comprehensively analyzed the genomic and functional attributes of the
H
30-Rx subclonal strains NA097 and NA114, belonging to the ST131 lineage. We carried out whole-genome sequencing, comparative analysis, phenotypic virulence assays, and profiling of the antibacterial responses of THP1 cells infected with these subclones. Phylogenomic analysis suggested that the strains were clonal in nature and confined entirely to a single clade. Comparative genomic analysis revealed that the virulence and resistance repertoires were comparable among the
H
30-Rx ST131 strains except for the commensal ST131 strain SE15. Similarly, seven phage-specific regions were found to be strongly associated with the
H
30-Rx strains but were largely absent in the genome of SE15. Phenotypic analysis confirmed the virulence and resistance similarities between the two strains. However, NA097 was found to be more robust than NA114 in terms of virulence gene carriage (
dra
operon), invasion ability (
P
< 0.05), and antimicrobial resistance (streptomycin resistance). RT
2
gene expression profiling revealed generic upregulation of key proinflammatory responses in THP1 cells, irrespective of ST131 lineage status. In conclusion, our study provides comprehensive, genome-inferred insights into the biology and immunological properties of ST131 strains and suggests clonal diversification of genomic and phenotypic features within the
H
30-Rx subclone of
E. coli
ST131.
Dual Zinc Transporter Systems in Vibrio cholerae Promote Competitive Advantages over Gut Microbiome
Authors: Sheng Ying , Fan Fenxia , Jensen Owen , Zhong Zengtao , Kan Biao , Wang Hui , Zhu Jun 
Published: July 2015

ABSTRACT

Zinc is an essential trace metal required for numerous cellular processes in all forms of life. In order to maintain zinc homeostasis, bacteria have developed several transport systems to regulate its uptake. In this study, we investigated zinc transport systems in the enteric pathogen
Vibrio cholerae
, the causative agent of cholera. Bioinformatic analysis predicts that two gene clusters, VC2081 to VC2083 (annotated as zinc utilization genes
znuABC
) and VC2551 to VC2555 (annotated as
z
inc-
r
egulated
g
enes
zrgABCDE
), are regulated by the putative zinc uptake regulator Zur. Using promoter reporter and biochemical assays, we confirmed that Zur represses
znuABC
and
zrgABCDE
promoters in a Zn
2+
-dependent manner. Under Zn
2+
-limiting conditions, we found that mutations in either the
znuABC
or
zrgABCDE
gene cluster affect bacterial growth, with
znuABC
mutants displaying a more severe growth defect, suggesting that both ZnuABC and ZrgABCDE are involved in Zn
2+
uptake and that ZnuABC plays the predominant role. Furthermore, we reveal that ZnuABC and ZrgABCDE are important for
V. cholerae
colonization in both infant and adult mouse models, particularly in the presence of other intestinal microbiota. Collectively, our studies indicate that these two zinc transporter systems play vital roles in maintaining zinc homeostasis during
V. cholerae
growth and pathogenesis.
Unexpected Roles for Toll-Like Receptor 4 and TRIF in Intraocular Infection with Gram-Positive Bacteria
Authors: Parkunan Salai Madhumathi , Randall C. Blake , Coburn Phillip S. , Astley Roger A. , Staats Rachel L. , Callegan Michelle C. 
Published: July 2015

ABSTRACTInflammation caused by infection with Gram-positive bacteria is typically initiated by interactions with Toll-like receptor 2 (TLR2). Endophthalmitis, an infection and inflammation of the posterior segment of the eye, can lead to vision loss when initiated by a virulent microbial pathogen. Endophthalmitis caused byBacillus cereusdevelops as acute inflammation with infiltrating neutrophils, and vision loss is potentially catastrophic. Residual inflammation observed duringB. cereusendophthalmitis in TLR2−/−mice led us to investigate additional innate pathways that may trigger intraocular inflammation. We first hypothesized that intraocular inflammation duringB. cereusendophthalmitis would be controlled by MyD88- and TRIF-mediated signaling, since MyD88 and TRIF are the major adaptor molecules for all bacterial TLRs. In MyD88−/−and TRIF−/−mice, we observed significantly less intraocular inflammation than in eyes from infected C57BL/6J mice, suggesting an important role for these TLR adaptors inB. cereusendophthalmitis. These results led to a second hypothesis, that TLR4, the only TLR that signals through both MyD88 and TRIF signaling pathways, contributed to inflammation duringB. cereusendophthalmitis. Surprisingly,B. cereus-infected TLR4−/−eyes also had significantly less intraocular inflammation than infected C57BL/6J eyes, indicating an important role for TLR4 inB. cereusendophthalmitis. Taken together, our results suggest that TLR4, TRIF, and MyD88 are important components of the intraocular inflammatory response observed in experimentalB. cereusendophthalmitis, identifying a novel innate immune interaction forB. cereusand for this disease.
Pseudomonas aeruginosa Quorum Sensing Molecule
N
-(3-Oxododecanoyl)-
l
-Homoserine-Lactone Induces HLA-G Expression in Human Immune Cells
Authors: Bortolotti Daria , LeMaoult Joel , Trapella Claudio , Di Luca Dario , Carosella Edgardo D. , Rizzo Roberta 
Published: July 2015

ABSTRACT

HLA-G is a nonclassical class I human leukocyte antigen (HLA) involved in mechanisms of immune tolerance. The objective of this study was to determine whether
N
-(3-oxododecanoyl)-
l
-homoserine lactone (3O-C
12
-HSL), a quorum sensing molecule produced by
Pseudomonas aeruginosa
, could modify HLA-G expression to control the host immune response. We evaluated the ability of 3O-C
12
-HSL to induce HLA-G expression in primary immune cells, monocytes (U937 and THP1), and T-cell lines (Jurkat)
in vitro
and analyzed the cellular pathway responsible for HLA-G expression. We studied the HLA-G promoter with a luciferase assay and interleukin-10 (IL-10) and p38/CREB signaling with enzyme-linked immunosorbent assay and immunofluorescence, respectively. We observed that 3O-C
12
-HSL is able to induce HLA-G expression in human monocytes and T cells. We showed that the induction of HLA-G by 3O-C
12
-HSL is p38/CREB and IL-10 dependent. 3O-C
12
-HSL treatment is able to arrest only the U937 cell cycle, possibly due to the peculiar expression of the ILT2 receptor in the U937 cell line. Our observations suggest HLA-G as a mechanism to create a protected niche for the bacterial reservoir, similar to the role of HLA-G molecules during viral infections.
Enzymatic Synthesis and Functional Characterization of Bioactive Microcin C-Like Compounds with Altered Peptide Sequence and Length
Authors: Bantysh Olga , Serebryakova Marina , Zukher Inna , Kulikovsky Alexey , Tsibulskaya Darya , Dubiley Svetlana , Severinov Konstantin 
Published: July 2015

ABSTRACT

Escherichia coli
microcin C (McC) consists of a ribosomally synthesized heptapeptide attached to a modified adenosine. McC is actively taken up by sensitive
Escherichia coli
strains through the YejABEF transporter. Inside the cell, McC is processed by aminopeptidases, which release nonhydrolyzable aminoacyl adenylate, an inhibitor of aspartyl-tRNA synthetase. McC is synthesized by the MccB enzyme, which terminally adenylates the MccA heptapeptide precursor MRTGNAN. Earlier, McC analogs with shortened peptide lengths were prepared by total chemical synthesis and were shown to have strongly reduced biological activity due to decreased uptake. Variants with longer peptides were difficult to synthesize, however. Here, we used recombinant MccB to prepare and characterize McC-like molecules with altered peptide moieties, including extended peptide lengths. We find that N-terminal extensions of
E. coli
MccA heptapeptide do not affect MccB-catalyzed adenylation and that some extended-peptide-length McC analogs show improved biological activity. When the peptide length reaches 20 amino acids, both YejABEF and SbmA can perform facilitated transport of toxic peptide adenylates inside the cell. A C-terminal fusion of the carrier maltose-binding protein (MBP) with the MccA peptide is also recognized by MccB
in vivo
and
in vitro
, allowing highly specific adenylation and/or radioactive labeling of cellular proteins.

IMPORTANCE
Enzymatic adenylation of chemically synthesized peptides allowed us to generate biologically active derivatives of the peptide-nucleotide antibiotic microcin C with improved bioactivity and altered entry routes into target cells, opening the way for development of various McC-based antibacterial compounds not found in nature.
Transposon Mutagenesis Paired with Deep Sequencing of Caulobacter crescentus under Uranium Stress Reveals Genes Essential for Detoxification and Stress Tolerance
Authors: Yung Mimi C. , Park Dan M. , Overton K. Wesley , Blow Matthew J. , Hoover Cindi A. , Smit John , Murray Sean R. , Ricci Dante P. , Christen Beat , Bowman Grant R. , Jiao Yongqin 
Published: July 2015

ABSTRACT

The ubiquitous aquatic bacterium
Caulobacter crescentus
is highly resistant to uranium (U) and facilitates U biomineralization and thus holds promise as an agent of U bioremediation. To gain an understanding of how
C. crescentus
tolerates U, we employed transposon (Tn) mutagenesis paired with deep sequencing (Tn-seq) in a global screen for genomic elements required for U resistance. Of the 3,879 annotated genes in the
C. crescentus
genome, 37 were found to be specifically associated with fitness under U stress, 15 of which were subsequently tested through mutational analysis. Systematic deletion analysis revealed that mutants lacking outer membrane transporters (

rsaF
a

and

rsaF
b

), a stress-responsive transcription factor (
cztR
), or a ppGpp synthetase/hydrolase (
spoT
) exhibited a significantly lower survival rate under U stress. RsaF
a
and RsaF
b
, which are homologues of TolC in
Escherichia coli
, have previously been shown to mediate S-layer export. Transcriptional analysis revealed upregulation of

rsaF
a

and

rsaF
b

by 4- and 10-fold, respectively, in the presence of U. We additionally show that

rsaF
a

mutants accumulated higher levels of U than the wild type, with no significant increase in oxidative stress levels. Our results suggest a function for RsaF
a
and RsaF
b
in U efflux and/or maintenance of membrane integrity during U stress. In addition, we present data implicating CztR and SpoT in resistance to U stress. Together, our findings reveal novel gene targets that are key to understanding the molecular mechanisms of U resistance in
C. crescentus
.

IMPORTANCE
Caulobacter crescentus
is an aerobic bacterium that is highly resistant to uranium (U) and has great potential to be used in U bioremediation, but its mechanisms of U resistance are poorly understood. We conducted a Tn-seq screen to identify genes specifically required for U resistance in
C. crescentus
. The genes that we identified have previously remained elusive using other omics approaches and thus provide significant insight into the mechanisms of U resistance by
C. crescentus
. In particular, we show that outer membrane transporters RsaF
a
and RsaF
b
, previously known as part of the S-layer export machinery, may confer U resistance by U efflux and/or by maintaining membrane integrity during U stress.