bluuebunny/crossref_metadata_2025 huggingface dataset:
Composite Bacterial Hopanoids and Their Microbial Producers across Oxygen Gradients in the Water Column of the California Current
Authors: Kharbush Jenan J. , Ugalde Juan A. , Hogle Shane L. , Allen Eric E. , Aluwihare Lihini I.
Published: September 2013

Hopanoids are pentacyclic triterpenoid lipids produced by many prokaryotes as cell membrane components. The structural variations of composite hopanoids, or bacteriohopanepolyols (BHPs), produced by various bacterial genera make them potentially useful molecular biomarkers of bacterial communities and metabolic processes in both modern and ancient environments. Building on previous work suggesting that organisms in low-oxygen environments are important contributors to BHP production in the marine water column and that there may be physiological roles for BHPs specific to these environments, this study investigated the relationship between trends in BHP structural diversity and abundance and the genetic diversity of BHP producers for the first time in a low-oxygen environment of the Eastern Tropical North Pacific. Amplification of the hopanoid biosynthesis gene for squalene hopene cyclase (
sqhC
) indicated far greater genetic diversity than would be predicted by examining BHP structural diversity alone and that greater
sqhC
genetic diversity exists in the marine environment than is represented by cultured representatives and most marine metagenomes. In addition, the genetic relationships in this data set suggest microaerophilic environments as potential “hot spots” of BHP production. Finally, structural analysis of BHPs showed that an isomer of the commonly observed BHP bacteriohopanetetrol may be linked to a producer that is more abundant in low-oxygen environments. Results of this study increase the known diversity of BHP producers and provide a detailed phylogeny with implications for the role of hopanoids in modern bacteria, as well as the evolutionary history of hopanoid biosynthesis, both of which are important considerations for future interpretations of the marine sedimentary record.
Use of a Hierarchical Oligonucleotide Primer Extension Approach for Multiplexed Relative Abundance Analysis of Methanogens in Anaerobic Digestion Systems
Authors: Wu Jer-Horng , Chuang Hui-Ping , Hsu Mao-Hsuan , Chen Wei-Yu
Published: September 2013

In this study, we established a rapid multiplex method to detect the relative abundances of amplified 16S rRNA genes from known cultivatable methanogens at hierarchical specificities in anaerobic digestion systems treating industrial wastewater and sewage sludge. The method was based on the hierarchical oligonucleotide primer extension (HOPE) technique and combined with a set of 27 primers designed to target the total archaeal populations and methanogens from 22 genera within 4 taxonomic orders. After optimization for their specificities and detection sensitivity under the conditions of multiple single-nucleotide primer extension reactions, the HOPE approach was applied to analyze the methanogens in 19 consortium samples from 7 anaerobic treatment systems (i.e., 513 reactions). Among the samples, the methanogen populations detected with order-level primers accounted for >77.2% of the PCR-amplified 16S rRNA genes detected using an
Archaea
-specific primer. The archaeal communities typically consisted of 2 to 7 known methanogen genera within the
Methanobacteriales
,
Methanomicrobiales
, and
Methanosarcinales
and displayed population dynamic and spatial distributions in anaerobic reactor operations. Principal component analysis of the HOPE data further showed that the methanogen communities could be clustered into 3 distinctive groups, in accordance with the distribution of the
Methanosaeta
,
Methanolinea
, and
Methanomethylovorans
, respectively. This finding suggested that in addition to acetotrophic and hydrogenotrophic methanogens, the methylotrophic methanogens might play a key role in the anaerobic treatment of industrial wastewater. Overall, the results demonstrated that the HOPE approach is a specific, rapid, and multiplexing platform to determine the relative abundances of targeted methanogens in PCR-amplified 16S rRNA gene products.
Cell Size Distributions of Soil Bacterial and Archaeal Taxa
Authors: Portillo Maria C. , Leff Jonathan W. , Lauber Christian L. , Fierer Noah
Published: September 2013

Cell size is a key ecological trait of soil microorganisms that determines a wide range of life history attributes, including the efficiency of nutrient acquisition. However, because of the methodological issues associated with determining cell sizes
in situ
, we have a limited understanding of how cell abundances vary across cell size fractions and whether certain microbial taxa have consistently smaller cells than other taxa. In this study, we extracted cells from three distinct soils and fractionated them into seven size ranges (5 μm to 0.2 μm) by filtration. Cell abundances in each size fraction were determined by direct microscopy, with the taxonomic composition of each size fraction determined by high-throughput sequencing of the 16S rRNA gene. Most of the cells were smaller than cells typically grown in culture, with 59 to 67% of cells <1.2 μm in diameter. Furthermore, each size fraction harbored distinct bacterial and archaeal communities in each of the three soils, and many of the taxa exhibited distinct size distribution patterns, with the smaller size fractions having higher relative abundances of taxa that are rare or poorly characterized (including
Acidobacteria
,
Gemmatimonadetes
,
Crenarchaeota
,
Verrucomicrobia
, and
Elusimicrobia
). In general, there was a direct relationship between average cell size and culturability, with those soil taxa that are poorly represented in culture collections tending to be smaller. Size fractionation not only provides important insight into the life history strategies of soil microbial taxa but also is a useful tool to enable more focused investigations into those taxa that remain poorly characterized.
Abilities of the mCP Agar Method and CRENAME Alpha Toxin-Specific Real-Time PCR Assay To Detect Clostridium perfringens Spores in Drinking Water
Authors: Maheux Andrée F. , Bérubé Ève , Boudreau Dominique K. , Villéger Romain , Cantin Philippe , Boissinot Maurice , Bissonnette Luc , Bergeron Michel G.
Published: September 2013

We first determined the analytical specificity and ubiquity (i.e., the ability to detect all or most strains) of a
Clostridium perfringens
-specific real-time PCR (rtPCR) assay based on the
cpa
gene (
cpa
rtPCR) by using a bacterial strain panel composed of
C. perfringens
and non-
C. perfringens Clostridium
strains. All non-
C. perfringens Clostridium
strains tested negative, whereas all
C. perfringens
strains tested positive with the
cpa
rtPCR, for an analytical specificity and ubiquity of 100%. The
cpa
rtPCR assay was then used to confirm the identity of 116 putative
C. perfringens
isolates recovered after filtration of water samples and culture on mCP agar. Colonies presenting discordant results between the phenotype on mCP agar and
cpa
rtPCR were identified by sequencing the 16S rRNA and
cpa
genes. Four mCP
−
/rtPCR
+
colonies were identified as
C. perfringens
, whereas 3 mCP
+
/rtPCR
−
colonies were identified as non-
C. perfringens
. The
cpa
rtPCR was negative with all 51 non-
C. perfringens
strains and positive with 64 of 65
C. perfringens
strains. Finally, we compared mCP agar and a CRENAME (
c
oncentration and
r
ecovery of microbial particles,
e
xtraction of
n
ucleic
a
cids, and
m
olecular
e
nrichment) procedure plus
cpa
rtPCR (CRENAME +
cpa
rtPCR) for their abilities to detect
C. perfringens
spores in drinking water. CRENAME +
cpa
rtPCR detected as few as one
C. perfringens
CFU per 100 ml of drinking water sample in less than 5 h, whereas mCP agar took at least 25 h to deliver results. CRENAME +
cpa
rtPCR also allows the simultaneous and sensitive detection of
Escherichia coli
and
C. perfringens
from the same potable water sample. In itself, it could be used to assess the public health risk posed by drinking water potentially contaminated with pathogens more resistant to disinfection.
DNA Double-Strand Break Repair at −15°C
Authors: Dieser Markus , Battista John R. , Christner Brent C.
Published: September 2013

The survival of microorganisms in ancient glacial ice and permafrost has been ascribed to their ability to persist in a dormant, metabolically inert state. An alternative possibility, supported by experimental data, is that microorganisms in frozen matrices are able to sustain a level of metabolic function that is sufficient for cellular repair and maintenance. To examine this experimentally, frozen populations of
Psychrobacter arcticus
273-4 were exposed to ionizing radiation (IR) to simulate the damage incurred from natural background IR sources in the permafrost environment from over ∼225 kiloyears (ky). High-molecular-weight DNA was fragmented by exposure to 450 Gy of IR, which introduced an average of 16 double-strand breaks (DSBs) per chromosome. During incubation at −15°C for 505 days,
P. arcticus
repaired DNA DSBs in the absence of net growth. Based on the time frame for the assembly of genomic fragments by
P. arcticus
, the rate of DNA DSB repair was estimated at 7 to 10 DSBs year
−1
under the conditions tested. Our results provide direct evidence for the repair of DNA lesions, extending the range of complex biochemical reactions known to occur in bacteria at frozen temperatures. Provided that sufficient energy and nutrient sources are available, a functional DNA repair mechanism would allow cells to maintain genome integrity and augment microbial survival in icy terrestrial or extraterrestrial environments.
Iterative Saturation Mutagenesis of −6 Subsite Residues in Cyclodextrin Glycosyltransferase from Paenibacillus macerans To Improve Maltodextrin Specificity for 2-
O
-
d
-Glucopyranosyl-
l
-Ascorbic Acid Synthesis
Authors: Han Ruizhi , Liu Long , Shin Hyun-dong , Chen Rachel R. , Li Jianghua , Du Guocheng , Chen Jian
Published: September 2013

2-
O
-
d
-Glucopyranosyl-
l
-ascorbic acid (AA-2G), a stable
l
-ascorbic acid derivative, is usually synthesized by cyclodextrin glycosyltransferase (CGTase), which contains nine substrate-binding subsites (from +2 to −7). In this study, iterative saturation mutagenesis (ISM) was performed on the −6 subsite residues (Y167, G179, G180, and N193) in the CGTase from
Paenibacillus macerans
to improve its specificity for maltodextrin, which is a cheap and easily soluble glycosyl donor for AA-2G synthesis. Site saturation mutagenesis of four sites—Y167, G179, G180, and N193—was first performed and revealed that four mutants—Y167S, G179R, N193R, and G180R—produced AA-2G yields higher than those of other mutant and wild-type CGTases. ISM was then conducted with the best positive mutant as a template. Under optimal conditions, mutant Y167S/G179K/N193R/G180R produced the highest AA-2G titer of 2.12 g/liter, which was 84% higher than that (1.15 g/liter) produced by the wild-type CGTase. Kinetics analysis of AA-2G synthesis using mutant CGTases confirmed the enhanced maltodextrin specificity and showed that compared to the wild-type CGTase, the mutants had no cyclization activity but high hydrolysis and disproportionation activities. A possible mechanism for the enhanced substrate specificity was also analyzed through structure modeling of the mutant and wild-type CGTases. These results indicated that the −6 subsite played crucial roles in the substrate binding and catalytic reactions of CGTase and that the obtained CGTase mutants, especially Y167S/G179K/N193R/G180R, are promising starting points for further development through protein engineering.
Stress Signaling Pathways for the Pathogenicity of Cryptococcus
Authors: Bahn Yong-Sun , Jung Kwang-Woo
Published: September 2013

Sensing, responding, and adapting to the surrounding environment are crucial for all living organisms to survive, proliferate, and differentiate in their biological niches. This ability is also essential for
Cryptococcus neoformans
and its sibling species
Cryptococcus gattii
, as these pathogens have saprobic and parasitic life cycles in natural and animal host environments. The ability of
Cryptococcus
to cause fatal meningoencephalitis is highly related to its capability to remodel and optimize its metabolic and physiological status according to external cues. These cues act through multiple stress signaling pathways through a panoply of signaling components, including receptors/sensors, small GTPases, secondary messengers, kinases, transcription factors, and other miscellaneous adaptors or regulators. In this minireview, we summarize and highlight the importance of several stress signaling pathways that influence the pathogenicity of
Cryptococcus
and discuss future challenges in these areas.
ICE
Slu
van, a 94-Kilobase Mosaic Integrative Conjugative Element Conferring Interspecies Transfer of VanB-Type Glycopeptide Resistance, a Novel Bacitracin Resistance Locus, and a Toxin-Antitoxin Stabilization System
Authors: Bjørkeng Eva K. , Hjerde Erik , Pedersen Torunn , Sundsfjord Arnfinn , Hegstad Kristin
Published: September 2013

A 94-kb integrative conjugative element (ICE
Slu
van) transferable to
Enterococcus faecium
and
Enterococcus faecalis
from an animal isolate of
Streptococcus lutetiensis
consists of a mosaic of genetic fragments from different Gram-positive bacteria. A variant of ICE
Slu
van was confirmed in
S. lutetiensis
from a patient. A complete Tn
5382
/Tn
1549
with a
vanB2
operon is integrated into a streptococcal ICE
Sde
3396-like region harboring a putative bacteriophage exclusion system, a putative agglutinin receptor precursor, and key components of a type IV secretion system. Moreover, ICE
Slu
van encodes a putative MobC family mobilization protein and a relaxase and, thus, in total has all genetic components essential for conjugative transfer. A 9-kb element within Tn
5382
/Tn
1549
encodes, among others, putative proteins similar to the TnpX site-specific recombinase in
Faecalibacterium
and VanZ in
Paenibacillus
, which may contribute to the detected low-level teicoplanin resistance. Furthermore, ICE
Slu
van encodes a novel bacitracin resistance locus that is associated with reduced susceptibility to bacitracin when transferred to
E. faecium
. The expression of a streptococcal
pezAT
toxin-antitoxin-encoding operon of ICE
Slu
van in
S. lutetiensis
,
E. faecium
, and
E. faecalis
was confirmed by reverse transcription (RT)-PCR, indicating an active toxin-antitoxin system which may contribute to stabilizing ICE
Slu
van within new hosts. Junction PCR and DNA sequencing confirmed that ICE
Slu
van excised to form a circular intermediate in
S. lutetiensis
,
E. faecalis
, and
E. faecium
. Transfer between
E. faecalis
cells was observed in the presence of helper plasmid pIP964. Sequence analysis of the original
S. lutetiensis
donor and enterococcal transconjugants showed that ICE
Slu
van integrates in a site-specific manner into the C-terminal end of the chromosomal tRNA methyltransferase gene
rumA
.
Advertising, Expectations and Informed Consent: The Contents and Functions of Acupuncture Leaflets
Authors: Bishop Felicity L , Salmon Cathy
Published: September 2013

Objective

To evaluate the content of patient information leaflets about acupuncture.

Methods

401 patient information leaflets were obtained from practising UK acupuncturists and subjected to content and thematic analysis.

Results

59% of included leaflets were from NHS physiotherapists. Almost all the leaflets defined acupuncture and the majority explained how it might work, described the treatment process and placed it in a historical context. Most described possible benefits and risks of acupuncture and discussed contraindications and safety. Just under a third of leaflets (120, 30%) suggested conditions that might be helped by acupuncture, most commonly musculoskeletal pain, arthritis and injuries. By emphasising differences between individuals in acupuncture treatments and responsiveness, the leaflets fostered hope for positive effects without making any guarantees.

Conclusions

Information leaflets are broadly consistent with the evidence for acupuncture, but some claims are inconsistent with official advice from advertising regulators. An ethically sound, scientifically grounded and psychologically effective leaflet should accurately convey both benefits and risks of treatment, optimise patients’ expectations and allay concerns about needling. This study suggests that acupuncture leaflets might achieve these multiple functions but care should be taken to ensure adequate coverage of risks.
Practical advice for avoidance of pain associated with insertion of intrauterine contraceptives
Authors: Bahamondes Luis , Mansour Diana , Fiala Christian , Kaunitz Andrew M , Gemzell-Danielsson Kristina
Published: September 2013

Few studies in the scientific literature provide clear direction on the prevention or management of pain associated with intrauterine contraceptive (IUC) placement. Those that have been published have studied small numbers of women and fail to provide definitive conclusions. There are also no guidelines available detailing recognised standard approaches to this problem. The consensus recommendations in this review focus primarily on non-pharmacological and often non-evidence-based interventions. This review includes general considerations, practical recommendations for both routine and more difficult cases and guidance on the optimal choice of instruments. General considerations, including pre-insertion counselling, the setting for the procedure, the confidence and technique of the provider and the interplay between the provider and assistant, can influence women's level of anxiety and, in turn, influence their perception of pain and their overall experience. Further studies are required to refine the optimal strategy for managing pain associated with IUC insertion.
Endovascular treatment of fusiform intracranial vertebral artery aneurysms using reconstructive techniques
Authors: Dabus Guilherme , Lin Eugene , Linfante Italo
Published: September 2013

Objective

To report our single center experience in the treatment of fusiform aneurysms involving the intracranial vertebral arteries using reconstructive endovascular techniques.

Patients and methods

The neurointerventional database of our institution was retrospectively reviewed from June 2010 to February 2013. Patients who underwent endovascular treatment of fusiform intracranial vertebral artery aneurysms using reconstructive techniques were included in the analysis. Clinical presentation, size, reconstructive technique used, procedural complication, and clinical and angiographic follow-ups were included in the analysis.

Results

Nine patients, aged 41–76 years (mean 54.8 years), were included. Mean angiographic diameter of unruptured aneurysms was 8.4 mm (range 4–14) while ruptured aneurysms averaged 6 mm (range 5–7 ). Two patients (two women) presented with acute subarachnoid hemorrhage (SAH). One patient with a large partially thrombosed aneurysm was treated with stent reconstruction requiring deployment of two stents (no coiling). There was one asymptomatic procedural complication (non-flow limiting cervical vertebral dissection). All patients had good clinical outcomes (modified Rankin Scale score of 0 or ≤1) including the two patients that presented with SAH (Hunt and Hess grades 2 and 3). There were no late hemorrhages at a mean clinical follow-up of 12 months (6–24 months). Eight patients had angiographic follow-up (6–18 months, mean 10.5 months) and six demonstrated aneurysm occlusion with complete vessel reconstruction at the angiographic follow-up.

Conclusions

The use of reconstructive techniques in the endovascular treatment of unruptured fusiform intracranial vertebral artery aneurysms is feasible, safe, and effective in the mid term. In patients presenting with SAH, however, the safety and effectiveness of these techniques remain unclear.
Levels of advertised unprotected vaginal and oral sex by independent indoor female sex workers in West Yorkshire, UK
Authors: Eccles Claire , Clarke Janette
Published: September 2013

Objectives

To assess the proportion of independent indoor female sex workers (FSW) in West Yorkshire, UK who advertise unprotected sex, and to investigate any association with cost, location and provision of anal sex.

Methods

Data on whether independent indoor FSW (defined as those not advertising via an escort agency or through a parlour) advertised unprotected sexual services, along with demographic data, were collected from 462 advertisement profiles of FSW in West Yorkshire from the websitehttp://www.adultwork.com. Independent t test and χ2statistics were used to test the association between advertised unprotected vaginal and oral sex, and FSW age, cost of services, location and whether they advertised anal sex.

Results

Unprotected vaginal sex was advertised by 8% of FSW, and unprotected oral sex by 74% of FSW. FSW advertising unprotected vaginal sex were more likely to live in Wakefield and Bradford than in Leeds, had significantly lower hourly rates, and were more likely to advertise anal sex.

Conclusions

Advertised condom use for vaginal and oral sex by independent indoor FSW in West Yorkshire was significantly lower than reported rates of protected sex found in previous studies based in London and the south of England. The advertisement of unprotected vaginal sex is associated with factors such as lower hourly rates and the advertisement of higher risk anal sex, which may signify greater economic need. FSW offering unprotected sex therefore represent an at-risk target group for health promotion.
Gaseous mercury flux measurements from two mine tailing sites in the Seal Harbour area of Nova Scotia
Authors: Dalziel J. , Tordon R.
Published: September 2013

During August 2004, mercury (Hg) flux-rates from surface soil were measured at an abandoned gold mining site near Seal Harbour, Nova Scotia. The Hg amalgamation process, used to isolate gold, produced areas of Hg-laden mine tailings. Diurnal measurements of the soil surface exchange of gaseous Hg to the atmosphere and meteorological data from two tailing deposit sites are discussed. These sites have different mining histories resulting in differing levels of Hg in the surface soil (0.05 µg g
−1
- Site 1, 23 µg g
−1
- Site 2). Site 1 showed a maximum flux-rate during mid-day at 60 ng m
−2
hr
−1
, declining to
c.
5 ng m
−2
hr
−1
during evening while Site 2 had a mid-day Hg flux-rate of 1292 ng m
−2
hr
−1
with levels of
c.
300 ng m
−2
hr
−1
during the evening. Mercury flux from each site has continuous diurnal Hg evasion at the soil surface. Correlations of Hg flux-rate with soil temperature, air temperature and solar radiation were evident at both sites and an Arrhenius-type relation of Hg flux with soil temperature was also evident. Data from these sites were compared to flux experiments conducted at impacted and other non-impacted sites in Nova Scotia.
Morphological and functional effects on cardiac tissue induced by moderate zinc deficiency during prenatal and postnatal life in male and female rats
Authors: Tomat Analia Lorena , Juriol Lorena Vanesa , Gobetto María Natalia , Veiras Luciana Cecilia , Mendes Garrido Abregú Facundo , Zilberman Judith , Fasoli Héctor , Elesgaray Rosana , Costa María Ángeles , Arranz Cristina Teresa
Published: September 2013

The aim of this study was to evaluate whether moderate zinc restriction in rats throughout fetal life, lactation, and/or postweaning growth results in early changes in cardiac morphology predisposing the onset of cardiac dysfunction in adult life as well as sex-related differences in the adaptation to this nutritional injury. Female Wistar rats received low or control zinc diets from the beginning of pregnancy up to offspring weaning. After being weaned, offspring were fed either a low or control zinc diet until 81 days. Systolic blood pressure was measured. Echocardiographic and electrocardiographic examinations, morphological experiments, and apoptosis by TUNEL assay were performed in the left ventricle. In the early stages, zinc-deficient male and female offspring showed an increase in cardiomyocyte diameter, probably associated with an increase in cardiac apoptotic cells, but smaller myocyte diameters in adulthood. In adult males, this nutritional injury induced decreased contractility and dilatation of the left ventricle, not allowing the heart to compensate the higher levels of blood pressure, and hypertrophic remodeling of coronary arteries associated with increased blood pressure. Adequate zinc intake during postweaning life did not overcome blood pressure levels but reversed some of the detrimental effects of earlier zinc deficiency in cardiac morphology and function. Females were less sensitive to this deficiency, exhibiting normal levels of blood pressure and no structural or functional heart alterations in adult life. The present study demonstrates that the effects of zinc deficiency on blood pressure, cardiac morphology, and function differ between sexes, with males more predisposed to develop cardiovascular diseases in adulthood.
Reduction of eotaxin production and eosinophil recruitment by pulmonary autologous macrophage transfer in a cockroach allergen-induced asthma model
Authors: Beal Dominic R. , Stepien David M. , Natarajan Sudha , Kim Jiyoun , Remick Daniel G.
Published: September 2013

We sought to investigate the effects of cockroach allergen (CRA) exposure on the lung macrophage population to determine how different macrophage phenotypes influence exacerbation of disease. CRA exposure caused significantly reduced expression of CD86 on lung macrophages. These effects were not systemic, as peritoneal macrophage CD86 expression was not altered. To investigate whether naïve macrophages could reduce asthma-like pulmonary inflammation, autologous peritoneal macrophages were instilled into the airways 24 h before the final CRA challenge. Pulmonary inflammation was assessed by measurement of airway hyperresponsiveness, mucin production, inflammatory cell recruitment, and cytokine production. Cell transfer did not have significant effects in control mice, nor did it affect pulmonary mucin production or airway hyperresponsiveness in control or CRA-exposed mice. However, there was significant reduction in the number of eosinophils recovered in the bronchoalveolar lavage (BAL) (5.8 × 105vs. 0.88 × 105), and total cell recruitment to the airways of CRA-exposed mice was markedly reduced (1.1 × 106vs. 0.57 × 106). The reduced eosinophil recruitment was reflected by substantially lower levels of eosinophil peroxidase in the lung and significantly lower concentrations of eotaxins in BAL (eotaxin 1: 3 pg/ml vs. undetectable; eotaxin 2: 2,383 vs. 131 pg/ml) and lung homogenate (eotaxin 1: 1,043 vs. 218 pg/ml; eotaxin 2: 10 vs. 1.5 ng/ml). We conclude that CRA decreases lung macrophage CD86 expression. Furthermore, supplementation of the lung cell population with peritoneal macrophages inhibits eosinophil recruitment, achieved through reduction of eotaxin production. These data demonstrate that transfer of naïve macrophages will reduce some aspects of asthma-like pulmonary inflammation in response to CRA.
PECAM-1 and caveolae form the mechanosensing complex necessary for NOX2 activation and angiogenic signaling with stopped flow in pulmonary endothelium
Authors: Noel John , Wang Hui , Hong Nankang , Tao Jian-Qin , Yu Kevin , Sorokina Elena M. , DeBolt Kristine , Heayn Michelle , Rizzo Victor , Delisser Horace , Fisher Aron B. , Chatterjee Shampa
Published: September 2013

We showed that stop of flow triggers a mechanosignaling cascade that leads to the generation of reactive oxygen species (ROS); however, a mechanosensor coupled to the cytoskeleton that could potentially transduce flow stimulus has not been identified. We showed a role for KATPchannel, caveolae (caveolin-1), and NADPH oxidase 2 (NOX2) in ROS production with stop of flow. Based on reports of a mechanosensory complex that includes platelet endothelial cell adhesion molecule-1 (PECAM-1) and initiates signaling with mechanical force, we hypothesized that PECAM-1 could serve as a mechanosensor in sensing disruption of flow. Using lungs in situ, we observed that ROS production with stop of flow was significantly reduced in PECAM-1−/−lungs compared with lungs from wild-type (WT) mice. Lack of PECAM-1 did not affect NOX2 activation machinery or the caveolin-1 expression or caveolae number in the pulmonary endothelium. Stop of flow in vitro triggered an increase in angiogenic potential of WT pulmonary microvascular endothelial cells (PMVEC) but not of PECAM-1−/−PMVEC. Obstruction of flow in lungs in vivo showed that the neutrophil infiltration as observed in WT mice was significantly lowered in PECAM-1−/−mice. With stop of flow, WT lungs showed higher expression of the angiogenic marker VEGF compared with untreated (sham) and PECAM-1−/−lungs. Thus PECAM-1 (and caveolae) are parts of the mechanosensing machinery that generates superoxide with loss of shear; the resultant ROS potentially drives neutrophil influx and acts as an angiogenic signal.
Efficient gene delivery to primary alveolar epithelial cells by nucleofection
Authors: Grzesik Benno A. , Vohwinkel Christine U. , Morty Rory E. , Mayer Konstantin , Herold Susanne , Seeger Werner , Vadász István
Published: September 2013

Primary alveolar epithelial cells play a pivotal role in lung research, particularly when focusing on gas exchange, barrier function, and transepithelial transport processes. However, efficient transfection of primary alveolar epithelial cells continues to be a major challenge. In the present study, we applied nucleofection, a novel method of gene and oligonucleotide delivery to the nucleus of cells by electroporation, to achieve highly efficient transfection of primary alveolar epithelial type II (ATII) cells. To quantify the amount of ATII cells effectively transfected, we applied a plasmid expressing GFP and assessed the amount of GFP-expressing cells by flow cytometry. Analysis of the nucleofected ATII cells revealed a concentration-dependent transfection efficiency of up to 50% when using 3–8 μg plasmid DNA without affecting cell viability. Nucleofection of cultured A549 and H441 cells yielded similar transfection rates. Importantly, nucleofection of ATII cells did not interfere with the integrity of ATII monolayers even with use of relatively high concentrations of plasmid DNA. In subsequent studies, we also efficiently delivered small interfering RNAs to ATII cells by nucleofection, thereby silencing Akt and the multiligand receptor megalin, which has been recently shown to play a key role in removal of excess protein from the alveolar space, and effectively inhibited megalin-driven uptake and transcellular transport of albumin in ATII cells. Thus we report successful transfection of primary rat alveolar epithelial cells with both plasmids and oligonucleotides via nucleofection with high viability and consistently good transfection rates without impairing key physiological properties of the cells.
Actin cytoskeleton redox proteome oxidation by cadmium
Authors: Go Young-Mi , Orr Michael , Jones Dean P.
Published: September 2013

Epidemiological studies associate environmental cadmium (Cd) exposure with the risk of lung diseases. Although mechanisms are not fully elucidated, several studies demonstrate Cd effects on actin and actin-associated proteins. In a recent study of Cd at concentrations similar to environmental exposures, we found that redox-dependent inflammatory signaling by NF-κB was sensitive to the actin-disrupting agent, cytochalasin D. The goal of the present study was to use mass spectrometry-based redox proteomics to investigate Cd effects on the actin cytoskeleton proteome and related functional pathways in lung cells at low environmental concentrations. The results showed that Cd under conditions that did not alter total protein thiols or glutathione redox state caused significant oxidation of peptidyl Cys of proteins regulating actin cytoskeleton. Immunofluorescence microscopy of lung fibroblasts and pulmonary artery endothelial cells showed that low-dose Cd exposure stimulated filamentous actin formation and nuclear localization of destrin, an actin-depolymerizing factor. Taken together, the results show that redox states of peptidyl Cys in proteins associated with actin cytoskeleton pathways are selectively oxidized in lung by Cd at levels thought to occur from environmental exposure.
RGS3 controls T lymphocyte migration in a model of Th2-mediated airway inflammation
Authors: Williams Jesse W. , Yau Douglas , Sethakorn Nan , Kach Jacob , Reed Eleanor B. , Moore Tamson V. , Cannon Judy , Jin Xiaohua , Xing Heming , Muslin Anthony J. , Sperling Anne I. , Dulin Nickolai O.
Published: September 2013

T cell migration toward sites of antigen exposure is mediated by G protein signaling and is a key function in the development of immune responses. Regulators of G protein signaling (RGS) proteins modulate G protein signaling; however, their role in the regulation of adaptive immune responses has not been thoroughly explored. Herein we demonstrated abundant expression of the Gi/Gq-specific RGS3 in activated T cells, and that diminished RGS3 expression in a T cell thymoma increased cytokine-induced migration. To examine the role of endogenous RGS3 in vivo, mice deficient in the RGS domain (RGS3ΔRGS) were generated and tested in an experimental model of asthma. Compared with littermate controls, the inflammation in the RGS3ΔRGSmice was characterized by increased T cell numbers and the striking development of perivascular lymphoid structures. Surprisingly, while innate inflammatory cells were also increased in the lungs of RGS3ΔRGSmice, eosinophil numbers and Th2 cytokine production were equivalent to control mice. In contrast, T cell numbers in the draining lymph nodes (dLN) were reduced in the RGS3ΔRGS, demonstrating a redistribution of T cells from the dLN to the lungs via increased RGS3ΔRGST cell migration. Together these novel findings show a nonredundant role for endogenous RGS3 in controlling T cell migration in vitro and in an in vivo model of inflammation.
Study on Characteristics of Chemokine CXCL10 Gene Cloned from cDNA Expression Library of Ujumqin Sheep
Authors: Hu P. F. , Li X. C. , Lei N. , Lan X. Y. , Zhao Q. J. , Guan W. J. , Ma Y. H.
Published: September 2013

Chemokines were a major regulator of body’s inflammatory and immune responses. In this study, the cDNA fragment of chemokine CXC ligand 10 (CXCL10) was cloned from the Ujumqin sheep ear marginal tissue cDNA expression library; the CXCL10 gene had 103 amino acids and a molecular weight of 11.47 kDa, and it shared a high homology among cattle, sheep, and goat, while a low homology compared with mouse. The CXCL10 protein had 4 conservative cysteine residues, located in 28, 30, 55, and 72 sites. The expression pattern and intracellular distribution of recombinant CXCL10 proteins in Ujumqin sheep fibroblast cells showed that there were green fluorescence signals both in cytoplasm and nucleolus after 24 h of transfection, the number of positive cells was increased with time, the peak level of fluorescence signal was reached after 48 h of transfection and the transfection efficiency was 33.3%; there was a significant decrease in fluorescence intensity after 72 h of transfection. Expression of recombinant CXCL10 gene inEscherichia colihad a time- and temperature-dependency on the amount of protein expression, and a small quantity of inducer was needed.
Solution of Point Reactor Neutron Kinetics Equations with Temperature Feedback by Singularly Perturbed Method
Authors: Chen Wenzhen , Hao Jianli , Chen Ling , Li Haofeng
Published: September 2013

The singularly perturbed method (SPM) is proposed to obtain the analytical solution for the delayed supercritical process of nuclear reactor with temperature feedback and small step reactivity inserted. The relation between the reactivity and time is derived. Also, the neutron density (or power) and the average density of delayed neutron precursors as the function of reactivity are presented. The variations of neutron density (or power) and temperature with time are calculated and plotted and compared with those by accurate solution and other analytical methods. It is shown that the results by the SPM are valid and accurate in the large range and the SPM is simpler than those in the previous literature.
Wellposedness of the Inverse Problem for Dirac Operator
Authors: Sat Murat , Panakhov Etibar S. , Tas Kezban
Published: September 2013

We study the wellposedness of the inverse problem for Dirac operator. We consider two different problems (unperturbed and perturbed problem) for Dirac operator, and then we prove that if the spectral characteristics of these problems are close to each other, then
the difference between their potential functions is sufficiently small.
A Best Proximity Point Result in Modular Spaces with the Fatou Property
Authors: Jleli Mohamed , Karapınar Erdal , Samet Bessem
Published: September 2013

Consider a nonself-mapping , where is a pair of nonempty subsets of a modular space . A best proximity point of is a point satisfying the condition: . In this paper, we introduce the class of proximal quasicontraction nonself-mappings in modular spaces with the Fatou property. For such mappings, we provide sufficient conditions assuring the existence and uniqueness of best proximity points.
Some Inclusion Relationships of Certain Subclasses of -Valent Functions Associated with a Family of Integral Operators
Authors: Aouf M. K. , El-Ashwah R. M. , Abd-Eltawab Ahmed M.
Published: September 2013

By making use of the new integral operator , we introduce and investigate several new subclasses of -valent starlike, -valent convex, -valent close-to-convex, and -valent quasi-convex functions. In particular, we establish some inclusion relationships associated with the aforementioned integral operators. Some of the results established in this paper would provide extensions of those given in earlier works.
Ear Acupressure for Smoking Cessation: A Randomised Controlled Trial
Authors: Zhang Anthony L. , Di Yuan Ming , Worsnop Christopher , May Brian H. , Da Costa Cliff , Xue Charlie C. L.
Published: September 2013

This study investigated the efficacy and safety of ear acupressure (EAP) as a stand-alone intervention for smoking cessation and the feasibility of this study design. Adult smokers were randomised to receive EAP specific for smoking cessation (SSEAP) or a nonspecific EAP (NSEAP) intervention which is not typically used for smoking cessation. Participants received 8 weekly treatments and were requested to press the five pellets taped to one ear at least three times daily. Participants were followed up for three months. Primary outcome measures were a 7-day point-prevalence cessation rate confirmed by exhaled carbon monoxide and relief of nicotine withdrawal symptoms (NWS). Intention-to-treat analysis was applied. Forty-three adult smokers were randomly assigned to SSEAP (n=20) or NSEAP (n=23) groups. The dropout rate was high with 19 participants completing the treatments and 12 remaining at followup. One participant from the SSEAP group had confirmed cessation at week 8 and end of followup (5%), but there was no difference between groups for confirmed cessation or NWS. Adverse events were few and minor.
Ethanol Extracts of FreshDavallia formosana(WL1101) Inhibit Osteoclast Differentiation by Suppressing RANKL-Induced Nuclear Factor-κB Activation
Authors: Lin Tzu-Hung , Yang Rong-Sen , Wang Kuan-Chin , Lu Dai-Hua , Liou Houng-Chi , Ma Yun , Chang Shao-Han , Fu Wen-Mei
Published: September 2013

The rhizome ofDavallia formosanais commonly used to treat bone disease including bone fracture, arthritis, and osteoporosis in Chinese herbal medicine. Here, we report the effects of WL1101, the ethanol extracts of fresh rhizomes ofDavallia formosanaon ovariectomy-induced osteoporosis. In addition, excess activated bone-resorbing osteoclasts play crucial roles in inflammation-induced bone loss diseases, including rheumatoid arthritis and osteoporosis. In this study, we examined the effects of WL1101 on receptor activator of nuclear factor-κB ligand (RANKL)-induced osteoclastogenesis. Treatment with WL1101 significantly inhibited RANKL-stimulated osteoclastogenesis. Two isolated active compounds, ((−)-epicatechin) or WL14 (4-hydroxy-3-aminobenzoic acid) could also inhibit RANKL-induced osteoclastogenesis. WL1101 suppressed the RANKL-induced nuclear factor-κB (NF-κB) activation and nuclear translocation, which is the key process during osteoclastogenesis, by inhibiting the activation of IκB kinase (IKK) and IκBα. In animal model, oral administration of WL1101 (50 or 200 mg/kg/day) effectively decreased the excess bone resorption and significantly antagonized the trabecular bone loss in ovariectomized rats. Our results demonstrate that the ethanol extracts of fresh rhizomes ofDavallia formosanainhibit osteoclast differentiation via the inhibition of NF-κB activation and effectively ameliorate ovariectomy-induced osteoporosis. WL1101 may thus have therapeutic potential for the treatment of diseases associated with excessive osteoclastic activity.
Finite Element Surface Registration Incorporating Curvature, Volume Preservation, and Statistical Model Information
Authors: Albrecht Thomas , Dedner Andreas , Lüthi Marcel , Vetter Thomas
Published: September 2013

We present a novel method for nonrigid registration of 3D surfaces and images. The method can be used to register surfaces by means of their distance images, or to register medical images directly. It is formulated as a minimization problem of a sum of several terms representing the desired properties of a registration result: smoothness, volume preservation, matching of the surface, its curvature, and possible other feature images, as well as consistency with previous registration results of similar objects, represented by a statistical deformation model. While most of these concepts are already known, we present a coherent continuous formulation of these constraints, including the statistical deformation model. This continuous formulation renders the registration method independent of its discretization. The finite element discretization we present is, while independent of the registration functional, the second main contribution of this paper. The local discontinuous Galerkin method has not previously been used in image registration, and it provides an efficient and general framework to discretize each of the terms of our functional. Computational efficiency and modest memory consumption are achieved thanks to parallelization and locally adaptive mesh refinement. This allows for the first time the use of otherwise prohibitively large 3D statistical deformation models.
Song Bu Li Decoction, a Traditional Uyghur Medicine, Protects Cell Death by Regulation of Oxidative Stress and Differentiation in Cultured PC12 Cells
Authors: Maiwulanjiang Maitinuer , Zhu Kevin Y. , Chen Jianping , Miernisha Abudureyimu , Xu Sherry L. , Du Crystal Y. Q. , Lau Kitty K. M. , Choi Roy C. Y. , Dong Tina T. X. , Aisa Haji A. , Tsim Karl W. K.
Published: September 2013

Song Bu Li decoction (SBL) is a traditional Uyghur medicinal herbal preparation, containing Nardostachyos Radix et Rhizoma. Recently, SBL is being used to treat neurological disorders (insomnia and neurasthenia) and heart disorders (arrhythmia and palpitation). Although this herbal extract has been used for many years, there is no scientific basis about its effectiveness. Here, we aimed to evaluate the protective and differentiating activities of SBL in cultured PC12 cells. The pretreatment of SBL protected the cell against tBHP-induced cell death in a dose-dependent manner. In parallel, SBL suppressed intracellular reactive oxygen species (ROS) formation. The transcriptional activity of antioxidant response element (ARE), as well as the key antioxidative stress proteins, was induced in dose-dependent manner by SBL in the cultures. In cultured PC12 cells, the expression of neurofilament, a protein marker for neuronal differentiation, was markedly induced by applied herbal extract. Moreover, the nerve growth factor- (NGF-) induced neurite outgrowth in cultured PC12 cells was significantly potentiated by the cotreatment of SBL. In accord, the expression of neurofilament was increased in the treatment of SBL. These results therefore suggested a possible role of SBL by its effect on neuron differentiation and protection against oxidative stress.
Drug Delivery Implants in the Treatment of Vitreous Inflammation
Authors: Wang Jillian , Jiang Angela , Joshi Malav , Christoforidis John
Published: September 2013

The eye is a model organ for the local delivery of therapeutics. This proves beneficial when treating vitreous inflammation and other ophthalmic pathologies. The chronicity of certain diseases, however, limits the effectiveness of locally administered drugs. To maintain such treatments often requires frequent office visits and can result in increased risk of infection and toxicity to the patient. This paper focuses on the implantable devices and particulate drug delivery systems that are currently being implemented and investigated to overcome these challenges. Implants currently on the market or undergoing clinical trials include those made of nonbiodegradable polymers, containing ganciclovir, fluocinolone acetonide, triamcinolone acetonide, and ranibizumab, and biodegradable polymers, containing dexamethasone, triamcinolone acetonide, and ranibizumab. Investigational intravitreal implants and particulate drug delivery systems, such as nanoparticles, microparticles, and liposomes, are also explored in this review article.
The Role of Natriuretic Peptides for the Diagnosis of Left Ventricular Dysfunction
Authors: Palazzuoli Alberto , Beltrami Matteo , Ruocco Gaetano , Pellegrini Marco , Nuti Ranuccio
Published: September 2013

Natriuretic peptides (NPs) are entered in current guidelines for heart failure (HF) diagnosis and management because of their high specificity and sensibility in screening patients with acute dyspnea. Due to their availability and relatively low cost, they became the first step examinations in HF patients evaluation at hospital admission together with clinical and chest radiography examination. NPs are released following any cardiac haemodynamic stress due to volume or pressure overload and should be considered as a mirror of cardiac condition helping in recognizing patients with poor outcome. Moreover, the exact role of NPs in early HF stages, in isolated diastolic dysfunction, and in general population is questioned. Several promising reports described their potential role; however, the wide cut‐off definition, inclusion criteria, and intrinsic measurement biases do not actually consent to their clinical application in these settings. A multimodality strategy including both NPs and imaging studies appears to be the best strategy to define the cardiac dysfunction etiology and its severity as well as to identify patients with higher risk. In this review, we describe the current and potential role of NPs in patients with asymptomatic cardiac insufficiency, evaluating the requirement to obtain a better standardization for imaging as for laboratory criteria.
Caristi Type Coincidence Point Theorem in Topological Spaces
Authors: Zhu Jiang , Wei Lei , Zhu Cheng-Cheng
Published: September 2013

A generalized Caristi type coincidence point theorem and its equivalences in the setting of topological spaces by using a kind of nonmetric type function are obtained. These results are used to establish variational principle and its equivalences ind-complete spaces, bornological vector space, seven kinds of completed quasi-semimetric spaces equipped withQ-functions, uniform spaces withq-distance, generating spaces of quasimetric family, and fuzzy metric spaces.