bluuebunny/crossref_metadata_2025_split huggingface dataset:
A group of grapevine MYBA transcription factors located in chromosome 14 control anthocyanin synthesis in vegetative organs with different specificities compared with the berry color locus
Authors: Matus José Tomás , Cavallini Erika , Loyola Rodrigo , Höll Janine , Finezzo Laura , Dal Santo Silvia , Vialet Sandrine , Commisso Mauro , Roman Federica , Schubert Andrea , Alcalde José Antonio , Bogs Jochen , Ageorges Agnès , Tornielli Giovanni Battista , Arce‐Johnson Patricio 
Published: April 2017

SummaryGrapevine organs accumulate anthocyanins in a cultivar‐specific and environmentally induced manner. The MYBA1‐A2 genes within the berry color locus in chromosome 2 represent the major genetic determinants of fruit color. The simultaneous occurrence of transposon insertions and point mutations in these genes is responsible for most white‐skinned phenotypes; however, the red pigmentation found in vegetative organs suggests the presence of additional regulators. This work describes a genomic region of chromosome 14 containing three closely related R2R3‐MYB genes, named MYBA5, MYBA6 and MYBA7. Ectopic expression of the latter two genes in grapevine hairy roots promoted anthocyanin accumulation without affecting other phenylpropanoids. Transcriptomic profiling of hairy roots expressing MYBA1, MYBA6 and MYBA7 showed that these regulators share the activation of late biosynthetic and modification/transport‐related genes, but differ in the activation of the FLAVONOID‐3′5′‐HYDROXYLASE (F3′5′H) family. An alternatively spliced MYBA6 variant was incapable of activating anthocyanin synthesis, however, because of the lack of an MYC1 interaction domain. MYBA1, MYBA6.1 and MYBA7 activated the promoters of UDP‐GLUCOSE:FLAVONOID 3‐O‐GLUCOSYLTRANSFERASE (UFGT) and ANTHOCYANIN 3‐O‐GLUCOSIDE‐6″‐O‐ACYLTRANSFERASE (3AT), but only MYBA1 induced F3′5′H in concordance with the low proportion of tri‐hydroxylated anthocyanins found in MYBA6‐A7 hairy roots. This putative new color locus is related to the red/cyanidic pigmentation of vegetative organs in black‐ and white‐skinned cultivars, and forms part of the UV‐B radiation response pathway orchestrated by ELONGATED HYPOCOTYL 5 (HY5). These results demonstrate the involvement of additional anthocyanin regulators in grapevine and suggest an evolutionary divergence between the two grape color loci for controlling additional targets of the flavonoid pathway.
Exudate collection using wound sponges—An easy, non‐invasive and reliable method to explore protease activities in ulcers
Authors: Ruf Marie‐Thérèse , Andreoli Arianna , Vujic Gabriel , Itin Peter , Pluschke Gerd , Schmid Peter 
Published: April 2017

AbstractProteases are important for wound healing, but in excessive amounts or left uncontrolled, they may cause healing impairment or other severe wound complications. Point‐of‐care testing for protease activities in wounds may be useful for monitoring the effectiveness of treatment, and for early identification of wounds that potentially fail to heal. Here we describe an easy, noninvasive method to collect wound fluid for evaluating the protease milieu of wounds. Wound fluids were collected using sterile sponges applied between wound surface and normal wound dressing. Wound fluid could be easily squeezed or centrifuged out of the sponges and was tested for gelatinase (MMP‐2 and MMP‐9) activities by gel zymography. In addition, we measured polymorphonuclear granulocyte elastase levels by ELISA. Both gelatinases were remarkably stable in sponge derived fluids, as no significant loss was observed even when samples were stored for 3 days at room temperature. Protease levels were highly diverse amongst patients and, in some cases, showed substantial variations in the course of the treatment. The here described wound sponge approach represents a patient‐friendly and reliable method to collect wound fluid for evaluating wound healing relevant biomarkers, such as matrix metalloproteinases.
Antibacterial activity of polihexanide formulations in a co‐culture of HaCaT keratinocytes and Staphylococcus aureus and at different pH levels
Authors: Wiegand Cornelia , Eberlein Thomas , Andriessen Anneke 
Published: April 2017

AbstractComplex stalled wounds feature an alkaline milieu that favors tissue destruction and microbial growth. The presence of bacteria in turn perpetuates the inflammatory response. However, only limited knowledge exists of pH dependency on the antibacterial efficacy of polyhexamethylene biguanide (PHMB) or the influence of surfactants or delivery vehicle used in antiseptic formulations. So far, PHMB alone has been shown to protect the keratinocytes from bacterial damage in such a co‐culture system as well as exhibiting increased antimicrobial activity at higher pH values. Here, the interaction of PHMB with the surfactants macrogolum and undecylenamidopropyl betaine that are most commonly used as additives in antiseptics and rinsing solutions such as Lavasept and Prontosan has been explored in addition to the PHMB‐containing biocellulose dressing Suprasorb X + PHMB. Undecylenamidopropyl betaine was found to lower the antimicrobial activity of polihexanide in the co‐culture system, while macrogolum and the biocellulose increased polihexanide efficiency to reduce Staphylococcus aureus especially in the presence of serum. The increasing antibacterial efficacy of PHMB with rising pH was not altered by undecylenamidopropyl betaine, macrogolum, or the biocellulose. The results suggest that application of PHMB with macrogolum or by delivery through a biocellulose dressing might be advantageous for management of wound infections.
Quality of life in patients with primary and secondary lymphedema in the community
Authors: Herberger Katharina , Blome Christine , Heyer Kristina , Ellis Franziska , Münter Karl‐Christian , Augustin Matthias 
Published: April 2017

AbstractLymphedema is a complex and burdensome medical problem and requires continuous specific therapy. The aim of this cross‐sectional study of community lymphedema care in the metropolitan area of Hamburg, Germany, was to evaluate health‐related quality of life (QoL) in lymphedema patients. Generic as well as disease‐specific health‐related QoL was assessed using EQ‐5D and FLQA‐LK, respectively. Pain was assessed using a visual analogue scale (VAS). About 301 patients (median age of 60.5 years, 90.8% female) with lymphedema of any origin were included. About 66.4% had lymphedema, 24.1% combined lipolymphedema, and 9.5% lipoedema. Mean disease‐specific QoL (FLQA‐LK) was 2.4 (range 0 = no to 4 = maximum burden). The highest impairment values were observed in subscales for physical complaints, everyday life, and emotional well‐being. Mean EQ‐5D VAS was 70.4, mean EQ‐5D score 63.3. Lymphedema was associated with major impairments in QoL, which differed for subgroups of pain, clinical severity, and comorbidity. Pain as a common problem for lymphedema patients seemed to be underestimated and undertreated. Early diagnosis and structured treatment strategies are urgently needed.
Cost‐effectiveness analysis of single‐use negative pressure wound therapy dressings (sNPWT) to reduce surgical site complications (SSC) in routine primary hip and knee replacements
Authors: Nherera Leo M. , Trueman Paul , Karlakki Sudheer L. 
Published: April 2017

AbstractWe sought to evaluate the cost‐effectiveness of single‐use negative pressure wound therapy in patients undergoing primary hip and knee replacements using effectiveness data from a recently completed non‐blinded randomized controlled trial. A decision analytic model was developed from UK National Health Service perspective using data from a single‐centre trial. 220 patients were randomized to treatment with either single‐use negative pressure wound therapy or standard care i.e., film dressings of clinician choice and followed for 6 weeks. Outcomes included dressing changes, length of stay, surgical site complications, cost and quality adjusted life years. The randomized controlled trial reported a reduction in dressing changes (p = 0.002), SSC (p = 0.06) and LOS (p = 0.07) in favor of single‐use negative pressure wound therapy compared with standard care. The model estimated 0.116 and 0.115 QALY gained, 0.98 and 0.92 complications avoided for single‐use negative pressure wound therapy and standard care, respectively. The cost/patient was £5,602 ($7,954) and £6,713 ($9,559) for single‐use negative pressure wound therapy and standard care respectively resulting in cost‐saving of £1,132 ($1,607) in favor of single‐use negative pressure wound therapy. Greater savings were observed in subgroups of higher risk patients with BMI ≥ 35 and ASA ≥ 3 i.e., £7,955 ($11,296) and £7,248 ($10,293), respectively. The findings were robust to a range of sensitivity analyses. In conclusion, single‐use negative pressure wound therapy can be considered a cost saving intervention to reduce surgical site complications following primary hip and knee replacements compared with standard care. Providers should consider targeting therapy to those patients at elevated risk of surgical site complications to maximize efficiency.
Development of a Novel Site-Specific Pegylated Interferon Beta for Antiviral Therapy of Chronic Hepatitis B Virus
Authors: Tsuge Masataka , Uchida Takuro , Hiraga Nobuhiko , Kan Hiromi , Makokha Grace Naswa , Abe-Chayama Hiromi , Miki Daiki , Imamura Michio , Ochi Hidenori , Hayes C. Nelson , Shimozono Rieko , Iwamura Tomokatsu , Narumi Hideki , Suzuki Tomohiko , Kainoh Mie , Taniguchi Tadatsugu , Chayama Kazuaki 
Published: April 2017

ABSTRACT

Although nucleot(s)ide analogues and pegylated interferon alpha 2a (PEG-IFN-α2a) can suppress hepatitis B virus (HBV) replication, it is difficult to achieve complete HBV elimination from hepatocytes. A novel site-specific pegylated recombinant human IFN-β (TRK-560) was recently developed. In the present study, we evaluated the antiviral effects of TRK-560 on HBV replication
in vitro
and
in vivo. In vitro
and
in vivo
HBV replication models were treated with antivirals including TRK-560, and changes in HBV markers were evaluated. To analyze antiviral mechanisms, cDNA microarray analysis and an enzyme-linked immunoassay (ELISA) were performed. TRK-560 significantly suppressed the production of intracellular HBV replication intermediates and extracellular HBV surface antigen (HBsAg) (
P
< 0.001 and
P
< 0.001, respectively), and the antiviral effects of TRK-560 were enhanced in combination with nucleot(s)ide analogues, such as entecavir and tenofovir disoproxil fumarate. The reduction in HBV DNA levels by TRK-560 treatment was significantly higher than that by PEG-IFN-α2a treatment both
in vitro
and
in vivo
(
P
= 0.004 and
P
= 0.046, respectively), and intracellular HBV covalently closed circular DNA (cccDNA) reduction by TRK-560 treatment was also significantly higher than that by PEG-IFN-α2a treatment
in vivo
(
P
= 0.0495). cDNA microarrays and ELISA for CXCL10 production revealed significant differences between TRK-560 and PEG-IFN-α2a in the induction potency of interferon-stimulated genes. TRK-560 shows a stronger antiviral potency via higher induction of interferon-stimulated genes and stronger stimulation of immune cell chemotaxis than PEG-IFN-α2a. As HBsAg loss and HBV cccDNA eradication are important clinical goals, these results suggest a potential role for TRK-560 in the development of more effective treatment for chronic hepatitis B infection.
High-Level Resistance of Staphylococcus aureus to β-Lactam Antibiotics Mediated by Penicillin-Binding Protein 4 (PBP4)
Authors: Hamilton Stephanie M. , Alexander J. Andrew N. , Choo Eun Ju , Basuino Li , da Costa Thaina M. , Severin Anatoly , Chung Marilyn , Aedo Sandra , Strynadka Natalie C. J. , Tomasz Alexander , Chatterjee Som S. , Chambers Henry F. 
Published: April 2017

ABSTRACT

Penicillin-binding protein 4 (PBP4), a nonessential, low-molecular-weight penicillin-binding protein of
Staphylococcus aureus
, has been implicated in low-level resistance to β-lactam antibiotics, although the mechanism is unknown. Mutations in PBP4 and its promoter were identified in a laboratory-generated mutant strain, CRB, which expresses high-level resistance to β-lactams, including resistance to the new-generation cephalosporins active against methicillin-resistant strains of
S. aureus
. These mutations did not appreciably alter the β-lactam antibiotic binding affinity of purified recombinant mutant PBP4 compared to that of wild-type PBP4. Compared to the susceptible parent strain, COLnex, the CRB strain produces a highly cross-linked cell wall peptidoglycan, indicative of increased transpeptidase activity. The
pbp4
promoter mutation of CRB was associated with greatly increased amounts of PBP4 in membranes compared to those in the COLnex parent. Replacement of the native promoter of COLnex with the mutant promoter of CRB resulted in increased amounts of PBP4 in membranes and a highly cross-linked cell wall. PBP4 can be repurposed to provide essential transpeptidase activity
in vivo
and confer high-level resistance to β-lactam antibiotics, such as ceftobiprole and ceftaroline.
Improved Xylose Metabolism by a
CYC8
Mutant of Saccharomyces cerevisiae
Authors: Nijland Jeroen G. , Shin Hyun Yong , Boender Leonie G. M. , de Waal Paul P. , Klaassen Paul , Driessen Arnold J. M. 
Published: April 2017

ABSTRACT

Engineering
Saccharomyces cerevisiae
for the utilization of pentose sugars is an important goal for the production of second-generation bioethanol and biochemicals. However,
S. cerevisiae
lacks specific pentose transporters, and in the presence of glucose, pentoses enter the cell inefficiently via endogenous hexose transporters (HXTs). By means of
in vivo
engineering, we have developed a quadruple hexokinase deletion mutant of
S. cerevisiae
that evolved into a strain that efficiently utilizes
d
-xylose in the presence of high
d
-glucose concentrations. A genome sequence analysis revealed a mutation (Y353C) in the general corepressor
CYC8
, or
SSN6
, which was found to be responsible for the phenotype when introduced individually in the nonevolved strain. A transcriptome analysis revealed altered expression of 95 genes in total, including genes involved in (i) hexose transport, (ii) maltose metabolism, (iii) cell wall function (mannoprotein family), and (iv) unknown functions (seripauperin multigene family). Of the 18 known HXTs, genes for 9 were upregulated, especially the low or nonexpressed
HXT10
,
HXT13
,
HXT15
, and
HXT16
. Mutant cells showed increased uptake rates of
d
-xylose in the presence of
d
-glucose, as well as elevated maximum rates of metabolism (
V
max
) for both
d
-glucose and
d
-xylose transport. The data suggest that the increased expression of multiple hexose transporters renders
d
-xylose metabolism less sensitive to
d
-glucose inhibition due to an elevated transport rate of
d
-xylose into the cell.

IMPORTANCE
The yeast
Saccharomyces cerevisiae
is used for second-generation bioethanol formation. However, growth on xylose is limited by pentose transport through the endogenous hexose transporters (HXTs), as uptake is outcompeted by the preferred substrate, glucose. Mutant strains were obtained with improved growth characteristics on xylose in the presence of glucose, and the mutations mapped to the regulator Cyc8. The inactivation of Cyc8 caused increased expression of HXTs, thereby providing more capacity for the transport of xylose, presenting a further step toward a more robust process of industrial fermentation of lignocellulosic biomass using yeast.
The Tale of a Neglected Energy Source: Elevated Hydrogen Exposure Affects both Microbial Diversity and Function in Soil
Authors: Khdhiri Mondher , Piché-Choquette Sarah , Tremblay Julien , Tringe Susannah G. , Constant Philippe 
Published: April 2017

ABSTRACT

The enrichment of H
2
-oxidizing bacteria (HOB) by H
2
generated by nitrogen-fixing nodules has been shown to have a fertilization effect on several different crops. The benefit of HOB is attributed to their production of plant growth-promoting factors, yet their interactions with other members of soil microbial communities have received little attention. Here we report that the energy potential of H
2
, when supplied to soil, alters ecological niche partitioning of bacteria and fungi, with multifaceted consequences for both generalist and specialist microbial functions. We used dynamic microcosms to expose soil to the typical atmospheric H
2
mixing ratio (0.5 ppmv) permeating soils, as well as mixing ratios comparable to those found at the soil-nodule interface (10,000 ppmv). Elevated H
2
exposure exerted direct effects on two HOB subpopulations distinguished by their affinity for H
2
while enhancing community level carbon substrate utilization potential and lowering CH
4
uptake activity in soil. We found that H
2
triggered changes in the abundance of microorganisms that were reproducible yet inconsistent across soils at the taxonomic level and even among HOB. Overall, H
2
exposure altered microbial process rates at an intensity that depends upon soil abiotic and biotic features. We argue that further examination of direct and indirect effects of H
2
on soil microbial communities will lead to a better understanding of the H
2
fertilization effect and soil biogeochemical processes.

IMPORTANCE
An innovative dynamic microcosm chamber system was used to demonstrate that H
2
diffusing in soil triggers changes in the distribution of HOB and non-HOB. Although the response was uneven at the taxonomic level, an unexpected coordinated response of microbial functions was observed, including abatement of CH
4
oxidation activity and stimulation of carbon turnover. Our work suggests that elevated H
2
rewires soil biogeochemical structure through a combination of direct effects on the growth and persistence of HOB and indirect effects on a variety of microbial processes involving HOB and non-HOB.
Reexamination of the Physiological Role of PykA in Escherichia coli Revealed that It Negatively Regulates the Intracellular ATP Levels under Anaerobic Conditions
Authors: Zhao Chunhua , Lin Zhao , Dong Hongjun , Zhang Yanping , Li Yin 
Published: April 2017

ABSTRACT

Pyruvate kinase is one of the three rate-limiting glycolytic enzymes that catalyze the last step of glycolysis, conversion of phosphoenolpyruvate (PEP) into pyruvate, which is associated with ATP generation. Two isozymes of pyruvate kinase, PykF and PykA, are identified in
Escherichia coli
. PykF is considered important, whereas PykA has a less-defined role. Prior studies inactivated the
pykA
gene to increase the level of its substrate, PEP, and thereby increased the yield of end products derived from PEP. We were surprised when we found a
pykA
::Tn
5
mutant in a screen for increased yield of an end product derived from pyruvate (
n
-butanol), suggesting that the role of PykA needs to be reexamined. We show that the
pykA
mutant exhibited elevated intracellular ATP levels, biomass concentrations, glucose consumption, and
n
-butanol production. We also discovered that the
pykA
mutant expresses higher levels of a presumed pyruvate transporter, YhjX, permitting the mutant to recapture and metabolize excreted pyruvate. Furthermore, we demonstrated that the nucleotide diphosphate kinase activity of PykA leads to negative regulation of the intracellular ATP levels. Taking the data together, we propose that inactivation of
pykA
can be considered a general strategy to enhance the production of pyruvate-derived metabolites under anaerobic conditions.

IMPORTANCE
This study showed that knocking out
pykA
significantly increased the intracellular ATP level and thus significantly increased the levels of glucose consumption, biomass formation, and pyruvate-derived product formation under anaerobic conditions.
pykA
was considered to be encoding a dispensable pyruvate kinase; here we show that
pykA
negatively regulates the anaerobic glycolysis rate through regulating the energy distribution. Thus, knocking out
pykA
can be used as a general strategy to increase the level of pyruvate-derived fermentative products.
Toxicity and Binding Studies of Bacillus thuringiensis Cry1Ac, Cry1F, Cry1C, and Cry2A Proteins in the Soybean Pests Anticarsia gemmatalis and Chrysodeixis (Pseudoplusia) includens
Authors: Bel Yolanda , Sheets Joel J. , Tan Sek Yee , Narva Kenneth E. , Escriche Baltasar 
Published: April 2017

ABSTRACT

Anticarsia gemmatalis
(velvetbean caterpillar) and
Chrysodeixis includens
(soybean looper, formerly named
Pseudoplusia includens
) are two important defoliating insects of soybeans. Both lepidopteran pests are controlled mainly with synthetic insecticides. Alternative control strategies, such as biopesticides based on the
Bacillus thuringiensis
(Bt) toxins or transgenic plants expressing Bt toxins, can be used and are increasingly being adopted. Studies on the insect susceptibilities and modes of action of the different Bt toxins are crucial to determine management strategies to control the pests and to delay outbreaks of insect resistance. In the present study, the susceptibilities of both soybean pests to the Bt toxins Cry1Ac, Cry1Fa, Cry1Ca, and Cry2Aa have been investigated. Bioassays performed in first-instar larvae showed that both insects are susceptible to all these toxins. Competition-binding studies carried out with Cry1Ac and Cry1Fa
125
-iodine labeled proteins demonstrated the presence of specific binding sites for both of them on the midgut brush border membrane vesicles (BBMVs) of both
A. gemmatalis
and
C. includens
. Competition-binding experiments and specific-binding inhibition studies performed with selected sugars and lectins indicated that Cry1Ac and Cry1Fa share some, but not all, binding sites in the midguts of both insects. Also, the Cry1Ac- or Cry1Fa-binding sites were not shared with Cry1Ca or Cry2Aa in either soybean pest. This study contributes to the knowledge of Bt toxicity and midgut toxin binding sites in
A. gemmatalis
and
C. includens
and sheds light on the cross-resistance potential of Cry1Ac, Cry1Fa, Cry1Ca, and Cry2Aa Bt proteins as candidate proteins for Bt-pyramided crops.

IMPORTANCE
In the present study, the toxicity and the mode of action of the
Bacillus thuringiensis
(Bt) toxins Cry1Ac, Cry1Fa, Cry1Ca, and Cry2Aa in
Anticarsia gemmatalis
and
Chrysodeixis includens
(important defoliating pests of soybeans) have been investigated. These studies are crucial for determining management strategies for pest control. Bioassays showed that both insects were susceptible to the toxins. Competition-binding studies demonstrated the presence of Cry1Fa- and Cry1Ac-specific binding sites in the midguts of both pests. These results, together with the results from binding inhibition studies performed with sugars and lectins, indicated that Cry1Ac and Cry1Fa share some, but not all, binding sites, and that they were not shared with Cry1Ca or Cry2Aa in either soybean pest. This study contributes to the knowledge of Bt toxicity in
A. gemmatalis
and
C. includens
and sheds light on the cross-resistance potential of Cry1Ac, Cry1Fa, Cry1Ca, and Cry2Aa Bt proteins as candidate proteins for Bt-pyramided crops.