bluuebunny/crossref_metadata_2025_split huggingface dataset:
Cathepsin B contributes to Na+ hyperabsorption in cystic fibrosis airway epithelial cultures
Authors: Tan Chong Da , Hobbs Carey , Sameni Mansoureh , Sloane Bonnie F. , Stutts M. Jackson , Tarran Robert 
Published: September 2014

Key points

The epithelial Na+ channel (ENaC) is hyperactive in cystic fibrosis (CF) airway epithelia, and contributes to excessive Na+ absorption and dehydration of the airway surface liquid (ASL) that lines the lungs – a major cause of CF lung disease.

The CF gene product (CFTR) is a cAMP‐regulated anion channel and how CFTR dysfunction contributes to ENaC hyperactivity is poorly understood. Of note, ENaC must be proteolytically cleaved to be active.

CF ASL is moderately acidic, due to the absence of HCO3− secretion through CFTR. The acid protease cathepsin B is present in the apical membrane of airway epithelia and is secreted into the ASL, suggesting that this protease may be active in the acidic CF environment.

We have found that cathepsin B induces activation of ENaC in both Xenopus laevis oocytes and in airway epithelia. Cathepsin B is also highly expressed in airway epithelia derived from patients with CF.

Inhibition of cathepsin B prevents Na+/ASL hyperabsorption in CF airway cultures, suggesting a hitherto unrecognized role for this protease in CF pathogenesis. Treatments directed at normalizing CF ASL pH or inhibiting cathepsin B may be useful in the treatment of CF lung disease.

AbstractIn cystic fibrosis (CF) lung disease, the absence of functional CF transmembrane conductance regulator results in Cl−/HCO3− hyposecretion and triggers Na+ hyperabsorption through the epithelial Na+ channel (ENaC), which contribute to reduced airway surface liquid (ASL) pH and volume. Prostasin, a membrane‐anchored serine protease with trypsin‐like substrate specificity has previously been shown to activate ENaC in CF airways. However, prostasin is typically inactive below pH 7.0, suggesting that it may be less relevant in acidic CF airways. Cathepsin B (CTSB) is present in both normal and CF epithelia and is secreted into ASL, but little is known about its function in the airways. We hypothesized that the acidic ASL seen in CF airways may stimulate CTSB to activate ENaC, contributing to Na+ hyperabsorption and depletion of CF ASL volume. In Xenopus laevis oocytes, CTSB triggered α‐ and γENaC cleavage and induced an increase in ENaC activity. In bronchial epithelia from both normal and CF donor lungs, CTSB localized to the apical membrane. In normal and CF human bronchial epithelial cultures, CTSB was detected at the apical plasma membrane and in the ASL. CTSB activity was significantly elevated in acidic ASL, which correlated with increased abundance of ENaC in the plasma membrane and a reduction in ASL volume. This acid/CTSB‐dependent activation of ENaC was ameliorated with the cell impermeable, CTSB‐selective inhibitor CA074, suggesting that CTSB inhibition may have therapeutic relevance. Taken together, our data suggest that CTSB is a pathophysiologically relevant protease that activates ENaC in CF airways.
Thiouracil-Forming Bacteria Identified and Characterized upon Porcine
In Vitro
Digestion of Brassicaceae Feed
Authors: Kiebooms Julie A. L. , Wauters Jella , Vanden Bussche Julie , Houf Kurt , De Vos Paul , Van Trappen Stefanie , Cleenwerck Ilse , Vanhaecke Lynn 
Published: September 2014

ABSTRACT

In recent years, the frequent detection of the banned thyreostat thiouracil (TU) in livestock urine has been related to endogenous TU formation following digestion of glucosinolate-rich Brassicaceae crops. Recently, it was demonstrated that, upon
in vitro
digestion of Brassicaceae, fecal bacteria induce TU detection in livestock (porcine livestock > bovines). Therefore, the present study was intended to isolate and identify bacteria involved in this intestinal TU formation upon Brassicaceae digestion and to gain more insight into the underlying mechanism in porcine livestock. Twenty porcine fecal inocula (gilts and multiparous sows) were assessed through static
in vitro
colonic-digestion simulations with rapeseed. After derivatization and extraction of the fecal suspensions, TU was analyzed using liquid chromatography-tandem mass spectrometry (LC-MS
2
). On average, lower TU concentrations were observed in fecal colonic simulations in gilts (8.35 ng g
−1
rapeseed ± 3.42 [mean ± standard deviation]) than in multiparous sows (52.63 ng g
−1
± 16.17), which correlates with maturation of the gut microbial population with age. Further exploration of the mechanism showed cell-dependent activity of the microbial conversion and sustained TU-forming activity after subjection of the fecal inoculum to moderate heat over a time span of up to 30 min. Finally, nine TU-producing bacterial species were successfully isolated and identified by a combination of biochemical and molecular techniques as
Escherichia coli
(
n
= 5),
Lactobacillus reuteri
(
n
= 2),
Enterococcus faecium
(
n
= 1), and
Salmonella enterica
subsp.
arizonae
(
n
= 1). This report demonstrates that endogenous formation of TU is Brassicaceae induced and occurs under colonic conditions most likely through myrosinase-like enzyme activity expressed by different common intestinal bacterial species.
Impact of Photocatalysis on Fungal Cells: Depiction of Cellular and Molecular Effects on Saccharomyces cerevisiae
Authors: Thabet Sana , Simonet France , Lemaire Marc , Guillard Chantal , Cotton Pascale 
Published: September 2014

ABSTRACT

We have investigated the antimicrobial effects of photocatalysis on the yeast model
Saccharomyces cerevisiae
. To accurately study the antimicrobial mechanisms of the photocatalytic process, we focused our investigations on two questions: the entry of the nanoparticles in treated cells and the fate of the intracellular environment. Transmission electronic microscopy did not reveal any entry of nanoparticles within the cells, even for long exposure times, despite degradation of the cell wall space and deconstruction of cellular compartments. In contrast to proteins located at the periphery of the cells, intracellular proteins did not disappear uniformly. Disappearance or persistence of proteins from the pool of oxidized intracellular isoforms was not correlated to their functions. Altogether, our data suggested that photocatalysis induces the establishment of an intracellular oxidative environment. This hypothesis was sustained by the detection of an increased level of superoxide ions (O
2
°
−
) in treated cells and by greater cell cultivability for cells expressing oxidant stress response genes during photocatalytic exposure. The increase in intracellular ROS, which was not connected to the entry of nanoparticles within the cells or to a direct contact with the plasma membrane, could be the result of an imbalance in redox status amplified by chain reactions. Moreover, we expanded our study to other yeast and filamentous fungi and pointed out that, in contrast to the laboratory model
S. cerevisiae
, some environmental strains are very resistant to photocatalysis. This could be related to the cell wall composition and structure.
Shared Binding Sites for the Bacillus thuringiensis Proteins Cry3Bb, Cry3Ca, and Cry7Aa in the African Sweet Potato Pest Cylas puncticollis (Brentidae)
Authors: Hernández-Martínez Patricia , Vera-Velasco Natalia Mara , Martínez-Solís María , Ghislain Marc , Ferré Juan , Escriche Baltasar 
Published: September 2014

ABSTRACT

Bacillus thuringiensis
Cry3Bb, Cry3Ca, and Cry7Aa have been reported to be toxic against larvae of the genus
Cylas
, which are important pests of sweet potato worldwide and particularly in sub-Saharan Africa. However, relatively little is known about the processing and binding interactions of these coleopteran-specific Cry proteins. The aim of the present study was to determine whether Cry3Bb, Cry3Ca, and Cry7Aa proteins have shared binding sites in
Cylas puncticollis
to orient the pest resistance strategy by genetic transformation. Interestingly, processing of the 129-kDa Cry7Aa protoxin using commercial trypsin or chymotrypsin rendered two fragments of about 70 kDa and 65 kDa. N-terminal sequencing of the trypsin-activated Cry7Aa fragments revealed that processing occurs at Glu
47
for the 70-kDa form or Ile
88
for the 65-kDa form. Homologous binding assays showed specific binding of the two Cry3 proteins and the 65-kDa Cry7Aa fragment to brush border membrane vesicles (BBMV) from
C. puncticollis
larvae. The 70-kDa fragment did not bind to BBMV. Heterologous-competition assays showed that Cry3Bb, Cry3Ca, and Cry7Aa (65-kDa fragment) competed for the same binding sites. Hence, our results suggest that pest resistance mediated by the alteration of a shared Cry receptor binding site might render all three Cry toxins ineffective.
Microfluidic Quantitative PCR for Simultaneous Quantification of Multiple Viruses in Environmental Water Samples
Authors: Ishii Satoshi , Kitamura Gaku , Segawa Takahiro , Kobayashi Ayano , Miura Takayuki , Sano Daisuke , Okabe Satoshi 
Published: September 2014

ABSTRACT
To secure food and water safety, quantitative information on multiple pathogens is important. In this study, we developed a microfluidic quantitative PCR (MFQPCR) system to simultaneously quantify 11 major human viral pathogens, including adenovirus, Aichi virus, astrovirus, enterovirus, human norovirus, rotavirus, sapovirus, and hepatitis A and E viruses. Murine norovirus and mengovirus were also quantified in our MFQPCR system as a sample processing control and an internal amplification control, respectively. River water contaminated with effluents from a wastewater treatment plant in Sapporo, Japan, was collected and used to validate our MFQPCR system for multiple viruses. High-throughput quantitative information was obtained with a quantification limit of 2 copies/μl of cDNA/DNA. Using this MFQPCR system, we could simultaneously quantify multiple viral pathogens in environmental water samples. The viral quantities obtained using MFQPCR were similar to those determined by conventional quantitative PCR. Thus, the MFQPCR system developed in this study can provide direct and quantitative information for viral pathogens, which is essential for risk assessments.
Bacilysin from Bacillus amyloliquefaciens FZB42 Has Specific Bactericidal Activity against Harmful Algal Bloom Species
Authors: Wu Liming , Wu Huijun , Chen Lina , Xie Shanshan , Zang Haoyu , Borriss Rainer , Gao Xuewen 
Published: September 2014

ABSTRACTHarmful algal blooms, caused by massive and exceptional overgrowth of microalgae and cyanobacteria, are a serious environmental problem worldwide.In the present study, we looked forBacillusstrains with sufficiently strong anticyanobacterial activity to be used as biocontrol agents. Among 24 strains,Bacillus amyloliquefaciensFZB42 showed the strongest bactericidal activity againstMicrocystis aeruginosa, with a kill rate of 98.78%. The synthesis of the anticyanobacterial substance did not depend on Sfp, an enzyme that catalyzes a necessary processing step in the nonribosomal synthesis of lipopeptides and polyketides, but was associated with thearogene cluster that is involved in the synthesis of thesfp-independent antibiotic bacilysin. Disruption ofbacB, the gene in the cluster responsible for synthesizing bacilysin, or supplementation with the antagonistN-acetylglucosamine abolished the inhibitory effect, but this was restored when bacilysin synthesis was complemented. Bacilysin caused apparent changes in the algal cell wall and cell organelle membranes, and this resulted in cell lysis. Meanwhile, there was downregulated expression ofglmS,psbA1,mcyB, andftsZ—genes involved in peptidoglycan synthesis, photosynthesis, microcystin synthesis, and cell division, respectively. In addition, bacilysin suppressed the growth of other harmful algal species. In summary, bacilysin produced byB. amyloliquefaciensFZB42 has anticyanobacterial activity and thus could be developed as a biocontrol agent to mitigate the effects of harmful algal blooms.
Structural Insights into 2,2′-Azino-Bis(3-Ethylbenzothiazoline-6-Sulfonic Acid) (ABTS)-Mediated Degradation of Reactive Blue 21 by Engineered Cyathus bulleri Laccase and Characterization of Degradation Products
Authors: Kenzom T. , Srivastava P. , Mishra S. 
Published: September 2014

ABSTRACT

Advanced oxidation processes are currently used for the treatment of different reactive dyes which involve use of toxic catalysts. Peroxidases are reported to be effective on such dyes and require hydrogen peroxide and/or metal ions.
Cyathus bulleri
laccase, expressed in
Pichia pastoris
, catalyzes efficient degradation (78 to 85%) of reactive azo dyes (reactive black 5, reactive orange 16, and reactive red 198) in the presence of synthetic mediator ABTS [2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)]. This laccase was engineered to degrade effectively reactive blue 21 (RB21), a phthalocyanine dye reported to be decolorized only by peroxidases. The 816-bp segment (toward the C terminus) of the
lcc
gene was subjected to random mutagenesis and enzyme variants (Lcc35, Lcc61, and Lcc62) were selected based on increased ABTS oxidizing ability. Around 78 to 95% decolorization of RB21 was observed with the ABTS-supplemented Lcc variants in 30 min. Analysis of the degradation products by mass spectrometry indicated the formation of several low-molecular-weight compounds. Mapping the mutations on the modeled structure implicated residues both near and far from the T1 Cu site that affected the catalytic efficiency of the mutant enzymes on ABTS and, in turn, the rate of oxidation of RB21. Several inactive clones were also mapped. The importance of geometry as well as electronic changes on the reactivity of laccases was indicated.
Ciprofloxacin Residues in Municipal Biosolid Compost Do Not Selectively Enrich Populations of Resistant Bacteria
Authors: Youngquist Caitlin P. , Liu Jinxin , Orfe Lisa H. , Jones Stephen S. , Call Douglas R. 
Published: September 2014

ABSTRACT

Biosolids and livestock manure are valuable high-carbon soil amendments, but they commonly contain antibiotic residues that might persist after land application. While composting reduces the concentration of extractable antibiotics in these materials, if the starting concentration is sufficiently high then remaining residues could impact microbial communities in the compost and soil to which these materials are applied. To examine this issue, ciprofloxacin was added to biosolid compost feedstock to achieve a total concentration of 19 ppm, approximately 5-fold higher than that normally detected by liquid chromatography-tandem mass spectrometry (LC-MS/MS) (1 to 3.5 ppm). This feedstock was placed into mesh bags that were buried in aerated compost bays. Once a week, a set of bags was removed and analyzed (treated and untreated, three replicates of each; 4 weeks). Addition of ciprofloxacin had no effect on the recovery of resistant bacteria at any time point (
P
= 0.86), and a separate bioassay showed that aqueous extractions from materials with an estimated 59 ppm ciprofloxacin had no effect on the growth of a susceptible strain of
Escherichia coli
(
P
= 0.28). Regression analysis showed that growth of the susceptible strain of
E. coli
can be reduced given a sufficiently high concentration of ciprofloxacin (
P
< 0.007), a result that is consistent with adsorption being the primary mechanism of sequestration. While analytical methods detected biologically significant concentrations of ciprofloxacin in the materials tested here, the culture-based methods were consistent with the materials having sufficient adsorptive capacity to prevent typical concentrations of ciprofloxacin residues from selectively enriching populations of resistant bacteria.
The burden of selected congenital anomalies amenable to surgery in low and middle-income regions: cleft lip and palate, congenital heart anomalies and neural tube defects
Authors: Higashi Hideki , Barendregt Jan J , Kassebaum Nicholas J , Weiser Thomas G , Bickler Stephen W , Vos Theo 
Published: September 2014

ObjectiveTo quantify the burden of selected congenital anomalies in low and middle-income countries (LMICs) that could be reduced should surgical programmes cover the entire population with access to quality care.DesignBurden of disease and epidemiological modelling.SettingLMICs from all global regions.PopulationAll prevalent cases of selected congenital anomalies at birth in 2010.Main outcome measuresDisability-adjusted life years (DALYs).Interventions and methodsSurgical programmes for three congenital conditions were analysed: clefts (lip and palate); congenital heart anomalies; and neural tube defects. Data from the Global Burden of Disease 2010 Study were used to estimate the combination of fatal burden that could be addressed by surgical care and the additional long-term non-fatal burden associated with increased survival.ResultsOf the estimated 21.6 million DALYs caused by these three conditions in LMICs, 12.4 million DALYs (57%) are potentially addressable by surgical care among the population born with such conditions. Neural tube defects have the largest potential with 76% of burden amenable by surgery, followed by clefts (59%) and congenital heart anomalies (49%). Sub-Saharan Africa and South Asia have the greatest proportion of surgically addressable burden for clefts (68%), North Africa and Middle East for congenital heart anomalies (73%), and South Asia for neural tube defects (81%).ConclusionsThere is an important and neglected role surgical programmes can play in reducing the burden of congenital anomalies in LMICs.
Has the incidence of empyema in Scottish children continued to increase beyond 2005?
Authors: Nath Stuart , Thomas Matt , Spencer David , Turner Steve 
Published: September 2014

BackgroundThe incidence of empyema increased dramatically in children during the 1990s and early 2000s. We investigated the relationship between changes in the incidence of childhood empyema in Scotland following the 2006 introduction of routine heptavalent conjugate pneumococcal vaccination (PCv-7) and the 2010 introduction of the 13-valent (PCV-13) vaccine.MethodsThis was a whole-population study of Scottish hospital admissions between 1981 and 2013 using ICD (International Classification of Diseases)-9 and ICD-10 diagnostic codes for empyema. The number of admissions for pneumonia and croup was also captured to give insight into secular trends in admissions with other related and unrelated respiratory presentations.ResultsThere were 217 admissions with empyema between 1981 and 2005 (mean incidence 9 cases/million/year) and 323 between 2006 and 2013 (mean incidence 47 cases/million/year), p<0.001. The introduction of conjugate vaccines in 2006 was associated with an overall increase in admissions for empyema of 2.0 (95% CI 1.4 to 2.8) per 100 000 children, however, the incidence rate ratio for empyema admission between 2010 and 2013 was lower relative to 2006–2009 (0.78 (95% CI 0.63 to 0.98)). Secular changes in pneumonia, but not croup, were comparable with those for empyema.ConclusionsThe incidence of empyema in Scottish children initially rose in children aged 1 to 9 years after the introduction of routine conjugate pneumococcal vaccination, however, empyema incidence has fallen since 2010 when the PCV-13 was introduced.
Validity and reliability of measurement of capillary refill time in children: a systematic review
Authors: Fleming Susannah , Gill Peter , Jones Caroline , Taylor James A , Van den Bruel Ann , Heneghan Carl , Thompson Matthew 
Published: September 2014

BackgroundMost guidelines recommend the use of capillary refill time (CRT) as part of the routine assessment of unwell children, but there is little consensus on the optimum method of measurement and cut-off time.MethodsWe searched Medline (from 1948), Embase (from 1980) and CINAHL (from 1991) to June 2014 to identify studies with information on the normal range of CRT in healthy children, the validity of CRT compared with reference standard measures of haemodynamic status, reliability and factors influencing measurement of CRT, such as body site, pressing time and temperature.FindingsWe included 21 studies on 1915 children. Four studies provided information on the relationship between CRT and measures of cardiovascular status, 13 provided data on the normal range of CRT, 7 provided data on reliability and 10 assessed the effect of various confounding factors. In children over 7 days of age, the upper limit of normal CRT is approximately 2 s when measured on a finger, and 4 s when measured on the chest or foot, irrespective of whether the child is feverish or not. Longer pressing times and ambient temperature outside 20°C–25°C are associated with longer CRT. Evidence suggests that the use of stopwatches reduces variability between observers.InterpretationWe recommend use of the following standardised CRT method of measurement: press on the finger for 5 s using moderate pressure at an ambient temperature of 20°C–25°C. A capillary refill time of 3 s or more should be considered abnormal.
Electronic risk assessment for venous thromboembolism: investigating physicians’ rationale for bypassing clinical decision support recommendations
Authors: Nwulu Ugochi , Brooks Hannah , Richardson Suzanna , McFarland Lorraine , Coleman Jamie J 
Published: September 2014

ObjectiveThe underutilisation of venous thromboembolism (VTE) prophylaxis is still a problem in the UK despite the emergence of national guidelines and incentives to increase the number of patients undergoing VTE risk assessments. Our objective was to examine the reasons doctors gave for not prescribing enoxaparin when recommended by an electronic VTE risk assessment alert.DesignWe used a qualitative research design to conduct a thematic analysis of free text entered into an electronic prescribing system.SettingThe study took place in a large University teaching hospital, which has a locally developed electronic prescribing system known as PICS (Prescribing, Information and Communication System).ParticipantsWe extracted prescription data from all inpatient admissions over a 7-month period in 2012 using the audit database of PICS.InterventionThe completion of the VTE risk assessment form introduced into the hospital-wide electronic prescribing and health records system is mandatory. Where doctors do not prescribe VTE prophylaxis when recommended, they are asked to provide a reason for this decision. The free-text field was introduced in May 2012.Primary and secondary outcome measuresFree-text reasons for not prescribing enoxaparin when recommended were thematically coded.ResultsA total of 1136 free-text responses from 259 doctors were collected in the time period and 1206 separate reasons were analysed and coded. 389 reasons (32.3%) for not prescribing enoxaparin were coded as being due to ‘clinical judgment’; in 288 (23.9%) of the responses, doctors were going to reassess the patient or prescribe enoxaparin; and in 245 responses (20.3%), the system was seen to have produced an inappropriate alert.ConclusionsIn order to increase specificity of warnings and avoid users developing alert fatigue, it is essential that an evaluation of user responses and/or end user feedback as to the appropriateness and timing of alerts is obtained.