bluuebunny/crossref_metadata_2025 huggingface dataset:
Influence of Gonadotrophin‐Induced First Oestrus on Gilt Fertility
Authors: Hidalgo DM , Friendship RM , Greiner L , Manjarín R , Amezcua MR , Domínguez JC , Kirkwood RN
Published: August 2014

Contents

The aim of this study was to determine the association between the oestrous response of pre‐pubertal gilts to gonadotrophin injection or boar exposure and their subsequent farrowing rate and litter size. At 154 days of age, randomly selected pre‐pubertal gilts received an intramuscular injection of 400 IU equine chorionic gonadotrophin plus 200 IU human chorionic gonadotrophin (PG600®; Merck Animal Health; n = 181). From the remaining pool of animals not treated with hormones, the first gilts showing signs of oestrus were selected to act as controls (n = 201). Boar exposure began at 155 days of age for both groups, and gilts were bred at a weight of approximately 130 kg. Comparisons were made between PG600®‐treated gilts exhibiting oestrus or not within 7 days post‐injection (early and late responders, respectively) and control gilts exhibiting oestrus or not within 30 days after beginning of boar exposure (select and non‐select control gilts, respectively). By 162 days, oestrus was detected in 67.5% of PG600®‐treated gilts compared with 5.7% of control gilts (p < 0.0001). The proportion of animals observed in oestrus at least three times before breeding was greater for select control gilts compared with early and late responder PG600®‐treated gilts (p ≤ 0.001). There were no significant differences in farrowing rate and litter size between the four treatment groups. These data indicate that PG600® is an effective tool to induce an earlier oestrus in gilts, that subsequent farrowing rate and born alive litter size compare favourably to that of select gilts and that gilts failing to respond promptly to hormonal stimulation do not exhibit compromised fertility.
Characteristics of patients with yellow nail syndrome and pleural effusion
Authors: Valdés Luis , Huggins John T. , Gude Francisco , Ferreiro Lucía , Álvarez‐Dobaño José M. , Golpe Antonio , Toubes María E. , González‐Barcala Francisco J. , José Esther San , Sahn Steven A.
Published: August 2014

Yellow nail syndrome (YNS) can be associated with a pleural effusion (PE) but the characteristics of these patients are not well defined. We performed a systematic review across four electronic databases for studies reporting clinical findings, PE characteristics, and most effective treatment of YNS. Case descriptions and retrospective studies were included, unrestricted by year of publication. We reviewed 112 studies (150 patients), spanning a period of nearly 50 years. The male/female ratio was 1.2/1. The median age was 60 years (range: 0–88). Seventy‐eight percent were between 41–80 years old. All cases had lymphoedema and 85.6% had yellow nails. PEs were bilateral in 68.3%. The appearance of the fluid was serous in 75.3%, milky in 22.3% and purulent in 3.5%. The PE was an exudate in 94.7% with lymphocytic predominance in 96% with a low count of nucleated cells. In 61 of 66 (92.4%) of patients, pleural fluid protein values were >3 g/dL, and typically higher than pleural fluid LDH. Pleurodesis and decortication/pleurectomy were effective in 81.8% and 88.9% of cases, respectively, in the treatment of symptomatic PEs. The development of YNS and PE occurs between the fifth to eighth decade of life and is associated with lymphoedema. The PE is usually bilateral and behaves as a lymphocyte‐predominant exudate. The most effective treatments appear to be pleurodesis and decortication/pleurectomy.
Bronchoscopic interventions for chronic obstructive pulmonary disease
Authors: Mineshita Masamichi , Slebos Dirk‐Jan
Published: August 2014

Over the past decade, several non‐surgical and minimally invasive bronchoscopic lung volume reduction (BLVR) techniques have been developed to treat patients with severe chronic obstructive pulmonary disease (COPD). BLVR can be significantly efficacious, suitable for a broad cohort of patients, and associated with a solid safety profile at a reasonable expense. The introduction of BLVR is also expected to accelerate the further development of interventional pulmonology worldwide. Recently, results from clinical studies on BLVR techniques have been published, providing valuable information about the procedure's indications, contraindications, patient‐selection criterion and outcomes. BLVR utilizing one‐way endobronchial valves is gaining momentum as an accepted treatment in regular medical practice because of the identification of best responders. Patients with a heterogeneous emphysema distribution and without inter‐lobar collateral ventilation show encouraging results. Furthermore, for patients with collateral ventilation, who are not considered candidates for valve treatment, and for patients with homogeneous emphysema, the introduction of lung volume reduction coil treatment is a promising solution. Moreover, with the development of newer treatment modalities, that is, biochemical sealant and thermal water vapor, the potential to treat emphysema irrespective of collateral flow, may be further increased. Nevertheless, patient selection for BLVR treatment will be crucial for the procedure's success and should be performed using a multidisciplinary team approach. Consequently, BLVR needs to be concentrated in high‐volume centres that will offer better quality and experience with treatment challenges and adverse events. This review gives a general overview of BLVR from an expert and scientific perspective.
Moving stem cell therapy to patients with idiopathic pulmonary fibrosis
Authors: Glassberg Marilyn K. , Toonkel Rebecca L.
Published: August 2014

See article, page 1013
Being human in early Virginia
Authors: Rome Alan S.
Published: August 2014

While there have been many important studies of English perceptions of the people they called ‘Indians’, all see, or at least imply, an unbridgeable chasm between the more positive descriptions and the seemingly more denigratory ones. Yet, the apparently vast differences between accounts actually converge under a more fundamental unity. Even if they sometimes disagreed over its exact implications, the English firmly agreed among themselves that the Indians were in fact fully human and rational and that the best proof of this was their conformity to the basic duties and precepts of the law of nature. Their often overstated disagreements were due to tensions within this understanding of humanity over the relative importance of reason and revelation for the affairs of the world. Regardless, from their early abortive ventures in Roanoke in the 1580s through to the so‐called ‘Massacre’ of 1622, the English enjoyed a consensus that the natives of Virginia had, at least in potential, a concept of the divine, language, civil society, and a reasonable amount of self‐control over their passions and appetites.
Safety assessment of jumps in ski racing
Authors: Schindelwig K. , Reichl W. , Kaps P. , Mössner M. , Nachbauer W.
Published: August 2014

The influence of important parameters on the flight trajectory for jumps in downhill World Cup races was investigated. To quantify the impact injury risk at landing, the parameter equivalent landing height (ELH) was introduced, which considered a variable slope inclination during the landing movement. Altogether, 145 runs at four different jumps in World Cup races and trainings were recorded and analyzed. A simulation model was developed to predict the flight phase of the skier. Drag and lift areas were selected by parameter identification to fit the simulation trajectory to the two‐dimensional data from the video analysis. The maximum values of the ELH which can be absorbed with muscle force was taken from the study of Minetti et al. for elite female and male ski racers. A sensitivity analysis based on the four jumps showed that ELH is mainly influenced by takeoff angle, takeoff speed, and the steepness of the landing surface. With the help of the developed simulation software, it should be possible to predict the ELH for jumps in advance. In case of an excessive ELH, improvements can be made by changing the takeoff inclination or the approach speed.
Making the Technological Transition – Citizens' Encounters with Digital Pension Services
Authors: Breit Eric , Salomon Robert
Published: August 2014

We examine in this article the impact of digitization on the provision of public services by studying how citizens experience the use of web portals in their pension planning. Based on focus groups and user test material, we elucidate five critical phases that each operate as an obstacle for citizens' further engagement in the digital self‐service process: interest, access, comprehension, reflection and support. We argue that these phases and the obstacles they entail illustrate a transition away from a situation in which control and agency over the quality of public services is embedded in a dyadic relationship between citizens and frontline personnel, and to a situation characterized by more complex relations between citizens, the front line, and the digital infrastructure. We argue that this transition implies that citizens are required to possess a new type of competence that contains both financial and digital skills. Citizens who are unable to develop or acquire such competence are likely to be disadvantaged by the services. Lastly, we argue that these developments pose significant challenges for public administrations to ensure the overall quality of the public services.
Surface free energy of the human skin and its critical surface tension of wetting in the skin/surfactant aqueous solution/air system
Authors: Krawczyk J.
Published: August 2014

Background/purpose

The purpose of these studies was to determine the surface free energy of the human skin and its critical surface tension of wetting in the skin – surfactant aqueous solution – air system in relation to different types of surfactants.

Methods

The surface free energy of the skin and its components was calculated using the equilibrium advancing contact angle values of water, formamide, and diiodomethane on the forearm skin surface. Next, taking into account the measured values of the contact angle of aqueous solutions of SDDS, CTAB, TX‐100 and TX‐114 on the skin surface and data of their surface tension, the critical surface tension of the skin wetting was determined.

Results

We can classify the skin surface as low‐energetic one. The critical surface tension of the skin wetting depends on the type of surfactant.

Conclusion

It is possible to determine the critical surface tension of the human skin wetting on the basis of the values of the contact angle of aqueous solutions of surfactants and their surface tension. In this respect, nonionic surfactants seem to be the most appropriate.
IMAGING DIAGNOSIS—ACUTE MESENTERIC ISCHEMIA ASSOCIATED WITH HYPERTROPHIC CARDIOMYOPATHY IN A CAT
Authors: Lee Miyoung , Park Nohwon , Kim Jaehwan , Kim Donghyun , Kim Hyunsoo , Eom Kidong
Published: August 2014

A middle‐aged cat was presented with vomiting, diarrhea, and pelvic limb paresis. Radiography showed cardiomegaly, interstitial pulmonary infiltration, distended intestinal loops, and portal venous gas. Hypertrophic cardiomyopathy (HCM) was confirmed. On CT, thrombi were identified at the abdominal aorta, superior mesenteric artery, bilateral renal arteries, and cauda vena cava at the level of the iliac vein bifurcation. Dilated and gas‐filled loops of bowel with a “paper thin wall” were observed on CT images and were considered indicative of necrotic bowel. This case report describes the CT findings of acute mesenteric ischemia associated with HCM in a cat.
Molecular bases of NMDA receptor subtype‐dependent properties
Authors: Glasgow Nathan G. , Siegler Retchless Beth , Johnson Jon W.
Published: August 2014

NMDA receptors (NMDARs) are a class of ionotropic glutamate receptors (iGluRs) that are essential for neuronal development, synaptic plasticity, learning and cell survival. Several features distinguish NMDARs from other iGluRs and underlie the crucial roles NMDARs play in nervous system physiology. NMDARs display slow deactivation kinetics, are highly Ca2+ permeable, and require depolarization to relieve channel block by external Mg2+ ions, thereby making them effective coincidence detectors. These properties and others differ among NMDAR subtypes, which are defined by the subunits that compose the receptor. NMDARs, which are heterotetrameric, commonly are composed of two GluN1 subunits and two GluN2 subunits, of which there are four types, GluN2A–D. ‘Diheteromeric’ NMDARs contain two identical GluN2 subunits. Gating and ligand‐binding properties (e.g. deactivation kinetics) and channel properties (e.g. channel block by Mg2+) depend strongly on the GluN2 subunit contained in diheteromeric NMDARs. Recent work shows that two distinct regions of GluN2 subunits control most diheteromeric NMDAR subtype‐dependent properties: the N‐terminal domain is responsible for most subtype dependence of gating and ligand‐binding properties; a single residue difference between GluN2 subunits at a site termed the GluN2 S/L site is responsible for most subtype dependence of channel properties. Thus, two structurally and functionally distinct regions underlie the majority of subtype dependence of NMDAR properties. This topical review highlights recent studies of recombinant diheteromeric NMDARs that uncovered the involvement of the N‐terminal domain and of the GluN2 S/L site in the subtype dependence of NMDAR properties.
Alport syndrome: its effects on the glomerular filtration barrier and implications for future treatment
Authors: Savige Judy
Published: August 2014

The glomerular filtration barrier comprises a fenestrated capillary endothelium, glomerular basement membrane and podocyte slit diaphragm. Over the past decade we have come to realise that permselectivity depends on size and not necessarily charge, that the molecular sieve depends on the podocyte contractile apparatus and is highly dynamic, and that protein uptake by proximal tubular epithelial cells stimulates signalling and the production of transcription factors and inflammatory mediators. Alport syndrome is the second commonest monogenic cause of renal failure after autosomal dominant polycystic kidney disease. Eighty per cent of patients have X‐linked disease caused by mutations in the COL4A5 gene. Most of these result in the replacement of the collagen IV α3α4α5 network with the α1α1α2 heterotrimer. Affected membranes also have ectopic laminin and increased matrix metalloproteinase levels, which makes them more susceptible to proteolysis. Mechanical stress, due to the less elastic membrane and hypertension, interferes with integrin‐mediated podocyte–GBM adhesion. Proteinuria occurs when urinary levels exceed tubular reabsorption rates, and initiates tubulointerstitial fibrosis. The glomerular mesangial cells produce increased TGFβ and CTGF which also contribute to glomerulosclerosis. Currently there is no specific therapy for Alport syndrome. However treatment with angiotensin converting enzyme (ACE) inhibitors delays renal failure progression by reducing intraglomerular hypertension, proteinuria, and fibrosis. Our greater understanding of the mechanisms underlying the GBM changes and their consequences in Alport syndrome have provided us with further novel therapeutic targets.
Novel determinants of the neuronal Cl− concentration
Authors: Delpire Eric , Staley Kevin J.
Published: August 2014

It is now a well‐accepted view that cation‐driven Cl− transporters in neurons are involved in determining the intracellular Cl− concentration. In the present review, we propose that additional factors, which are often overlooked, contribute substantially to the Cl− gradient across neuronal membranes. After briefly discussing the data supporting and opposing the role of cation–chloride cotransporters in regulating Cl−, we examine the participation of the following factors in the formation of the transmembrane Cl− gradient: (i) fixed ‘Donnan’ charges inside and outside the cell; (ii) the properties of water (free vs. bound); and (iii) water transport through the cotransporters. We demonstrate a steep relationship between intracellular Cl− and the concentration of fixed negative charges on macromolecules. We show that in the absence of water transport through the K+–Cl− cotransporter, a large osmotic gradient builds at concentrations below or above a set value of ‘Donnan’ charges, and show that at any value of these fixed charges, the reversal potential for Cl− equates that of K+. When the movement of water across the membrane is a source of free energy, it is sufficient to modify the movement of Cl− through the cotransporter. In this scenario, the reversal potential for Cl− does not closely follow that of K+. Furthermore, our simulations demonstrate that small differences in the availability of freely diffusible water between inside and outside the cell greatly affect the Cl− reversal potential, particularly when osmolar transmembrane gradients are minimized, for example by idiogenic osmoles. We also establish that the presence of extracellular charges has little effect on the chloride reversal potential, but greatly affects the effective inhibitory conductance for Cl−. In conclusion, our theoretical analysis of the presence of fixed anionic charges and water bound on macromolecules inside and outside the cell greatly impacts both Cl− gradient and Cl− conductance across neuronal membranes.
Serotonin neurones have anti‐convulsant effects and reduce seizure‐induced mortality
Authors: Buchanan Gordon F. , Murray Nicholas M. , Hajek Michael A. , Richerson George B.
Published: August 2014

Key points

Sudden unexpected death in epilepsy is the leading cause of death in patients with refractory epilepsy.

Respiratory and cardiac impairment induced by a seizure have been identified as possible causes of seizure‐related death, but which is more important has been the subject of debate.

Serotonin has been linked to seizure control, but whether it is primarily anti‐convulsant or proconvulsant remains controversial.

In this study we induced seizures in mice with a genetic deletion of serotonin neurones and their phenotypically normal littermates while recording EEG, EMG, ECG and breathing, and assessed the effects of seizures on breathing, cardiac activity and survival

Serotonin and serotonin neurones are involved in setting the seizure threshold, regulating seizure severity and preventing mortality, and death in at least one seizure model is due to respiratory arrest, which can be prevented with selective serotonin reuptake inhibitor treatment or 5‐HT2A receptor activation.

Sudden unexpected death in epilepsy (SUDEP) is the leading cause of death in patients with refractory epilepsy. Defects in central control of breathing are important contributors to the pathophysiology of SUDEP, and serotonin (5‐HT) system dysfunction may be involved. Here we examined the effect of 5‐HT neurone elimination or 5‐HT reduction on seizure risk and seizure‐induced mortality. Adult Lmx1bf/f/p mice, which lack >99% of 5‐HT neurones in the CNS, and littermate controls (Lmx1bf/f) were subjected to acute seizure induction by maximal electroshock (MES) or pilocarpine, variably including electroencephalography, electrocardiography, plethysmography, mechanical ventilation or pharmacological therapy. Lmx1bf/f/p mice had a lower seizure threshold and increased seizure‐induced mortality. Breathing ceased during most seizures without recovery, whereas cardiac activity persisted for up to 9 min before terminal arrest. The mortality rate of mice of both genotypes was reduced by mechanical ventilation during the seizure or 5‐HT2A receptor agonist pretreatment. The selective serotonin reuptake inhibitor citalopram reduced mortality of Lmx1bf/f but not of Lmx1bf/f/p mice. In C57BL/6N mice, reduction of 5‐HT synthesis with para‐chlorophenylalanine increased MES‐induced seizure severity but not mortality. We conclude that 5‐HT neurones raise seizure threshold and decrease seizure‐related mortality. Death ensued from respiratory failure, followed by terminal asystole. Given that SUDEP often occurs in association with generalised seizures, some mechanisms causing death in our model might be shared with those leading to SUDEP. This model may help determine the relationship between seizures, 5‐HT system dysfunction, breathing and death, which may lead to novel ways to prevent SUDEP.
Influence of heat treatments upon the mechanical properties and in vitro bioactivity of ZrO2-toughened MgO-CaO-SiO2-P2O5-CaF2 glass–ceramics
Authors: Li Huan-cai , Wang Dian-gang , Meng Xiang-guo , Chen Chuan-zhong
Published: August 2014

Zirconia-toughened MgO-CaO-SiO2-P2O5-CaF2 glass–ceramics are prepared using sintering techniques, and a series of heat treatment procedures are designed to obtain a glass–ceramic with improved properties. The crystallization behavior, phase composition, and morphology of the glass–ceramics are characterized. The bending strength, elastic modulus, fracture toughness, and microhardness of the glass–ceramics are investigated, and the effect mechanism of heat treatments upon the mechanical properties is discussed. The bioactivity of glass–ceramics is then evaluated using the in vitro simulated body fluid (SBF) soaking test, and the mechanism whereby apatite forms on the glass–ceramic surfaces in the SBF solution is discussed. The results indicate that the main crystal phase of the G-24 sample undergoing two heat treatment procedures is Ca5(PO4)3F (fluorapatite), and those of the G-2444 sample undergoing four heat treatment procedures are Ca5(PO4)3F and β-CaSiO3 (β-wollastonite). The heat treatment procedures are found to greatly influence the mechanical properties of the glass–ceramic, and an apatite layer is induced on the glass–ceramic surface after soaking in the SBF solution.
Hydrogen induced electric conduction in undoped ZnO and Ga-doped ZnO thin films: Creating native donors via reduction, hydrogen donors, and reactivating extrinsic donors
Authors: Akazawa Housei
Published: August 2014

The manner in which hydrogen atoms contribute to the electric conduction of undoped ZnO and Ga-doped ZnO (GZO) films was investigated. Hydrogen atoms were permeated into these films through annealing in an atmospheric H2 ambient. Because the creation of hydrogen donors competes with the thermal annihilation of native donors at elevated temperatures, improvements to electric conduction from the initial state can be observed when insulating ZnO films are used as samples. While the resistivity of conductive ZnO films increases when annealing them in a vacuum, the degree of increase is mitigated when they are annealed in H2. Hydrogenation of ZnO crystals was evidenced by the appearance of OH absorption signals around a wavelength of 2700 nm in the optical transmittance spectra. The lowest resistivity that was achieved by H2 annealing was limited to 1–2 × 10−2 Ω cm, which is one order of magnitude higher than that by native donors (2–3 × 10−3 Ω cm). Hence, all native donors are converted to hydrogen donors. In contrast, GZO films that have resistivities yet to be improved become more conductive after annealing in H2 ambient, which is in the opposite direction of GZO films that become more resistive after vacuum annealing. Hydrogen atoms incorporated into GZO crystals should assist in reactivating Ga3+ donors.
Reducing sewer corrosion through integrated urban water management
Authors: Pikaar Ilje , Sharma Keshab R. , Hu Shihu , Gernjak Wolfgang , Keller Jürg , Yuan Zhiguo
Published: August 2014

Sourcing corrosive sewer sulfides

Sewer systems are corroding at an alarming rate, costing governments billions of dollars to replace. Differences among water treatment systems make it difficult to track down the source of corrosive sulfide responsible for this damage. Pikaar
et al.
performed an extensive industry survey and sampling campaign across Australia (see the Perspective by Rauch and Kleidorfer). Aluminum sulfate added as a coagulant during drinking water treatment was the primary culprit in corroding sewer systems. Modifying this common treatment strategy to include sulfate-free coagulants could dramatically reduce sewer corrosion across the globe.

Science
, this issue p.
812
; see also p.
734
Preterm labor: One syndrome, many causes
Authors: Romero Roberto , Dey Sudhansu K. , Fisher Susan J.
Published: August 2014

Preterm birth is associated with 5 to 18% of pregnancies and is a leading cause of infant morbidity and mortality. Spontaneous preterm labor, a syndrome caused by multiple pathologic processes, leads to 70% of preterm births. The prevention and the treatment of preterm labor have been long-standing challenges. We summarize the current understanding of the mechanisms of disease implicated in this condition and review advances relevant to intra-amniotic infection, decidual senescence, and breakdown of maternal-fetal tolerance. The success of progestogen treatment to prevent preterm birth in a subset of patients at risk is a cause for optimism. Solving the mystery of preterm labor, which compromises the health of future generations, is a formidable scientific challenge worthy of investment.
The biology of mammalian parenting and its effect on offspring social development
Authors: Rilling James K. , Young Larry J.
Published: August 2014

Parents know the transformative nature of having and caring for a child. Among many mammals, giving birth leads from an aversion to infant stimuli to irresistible attraction. Here, we review the biological mechanisms governing this shift in parental motivation in mammals. Estrogen and progesterone prepare the uterus for embryo implantation and placental development. Prolactin stimulates milk production, whereas oxytocin initiates labor and triggers milk ejection during nursing. These same molecules, interacting with dopamine, also activate specific neural pathways to motivate parents to nurture, bond with, and protect their offspring. Parenting in turn shapes the neural development of the infant social brain. Recent work suggests that many of the principles governing parental behavior and its effect on infant development are conserved from rodent to humans.
Large environments reveal the statistical structure governing hippocampal representations
Authors: Rich P. Dylan , Liaw Hua-Peng , Lee Albert K.
Published: August 2014

Nerve cells displaying extra large spaces

Rats use brain cells called “place” cells to figure out where they are. Richet al.used mazes or tracks many meters long—the size of rats' ranges in the wild—to investigate how rats represent a very large environment or extended experience in their brain. As novel environments became larger and larger, the rats' brains recruited new place cells. However, no matter how large the environment became, some cells always remained silent, perhaps as a reserve for other environments yet to be visited.

Science, this issue p.814
Nature's first functional food
Authors: Gura Trisha
Published: August 2014

Breast milk feeds helpful microbes, fights harmful ones, provides immunity, and jump-starts a newborn's life.
Vibrio harveyi Adheres to and Penetrates Tissues of the European Abalone Haliotis tuberculata within the First Hours of Contact
Authors: Cardinaud Marion , Barbou Annaïck , Capitaine Carole , Bidault Adeline , Dujon Antoine Marie , Moraga Dario , Paillard Christine
Published: August 2014

Vibrio harveyi
is a marine bacterial pathogen responsible for episodic epidemics generally associated with massive mortalities in many marine organisms, including the European abalone
Haliotis tuberculata
. The aim of this study was to identify the portal of entry and the dynamics of infection of
V. harveyi
in the European abalone. The results indicate that the duration of contact between
V. harveyi
and the European abalone influences the mortality rate and precocity. Immediately after contact, the epithelial and mucosal area situated between the gills and the hypobranchial gland was colonized by
V. harveyi
. Real-time PCR analyses and culture quantification of a green fluorescent protein-tagged strain of
V. harveyi
in abalone tissues revealed a high density of bacteria adhering to and then penetrating the whole gill-hypobranchial gland tissue after 1 h of contact.
V. harveyi
was also detected in the hemolymph of a significant number of European abalones after 3 h of contact. In conclusion, this article shows that a TaqMan real-time PCR assay is a powerful and useful technique for the detection of a marine pathogen such as
V. harveyi
in mollusk tissue and for the study of its infection dynamics. Thus, we have revealed that the adhesion and then the penetration of
V. harveyi
in European abalone organs begin in the first hours of contact. We also hypothesize that the portal of entry of
V. harveyi
in the European abalone is the area situated between the gills and the hypobranchial gland.
Bacteriophage ϕMAM1, a Viunalikevirus, Is a Broad-Host-Range, High-Efficiency Generalized Transducer That Infects Environmental and Clinical Isolates of the Enterobacterial Genera Serratia and Kluyvera
Authors: Matilla Miguel A. , Salmond George P. C.
Published: August 2014

Members of the enterobacterial genus
Serratia
are ecologically widespread, and some strains are opportunistic human pathogens. Bacteriophage ϕMAM1 was isolated on
Serratia plymuthica
A153, a biocontrol rhizosphere strain that produces the potently bioactive antifungal and anticancer haterumalide oocydin A. The ϕMAM1 phage is a generalized transducing phage that infects multiple environmental and clinical isolates of
Serratia
spp. and a rhizosphere strain of
Kluyvera cryocrescens
. Electron microscopy allowed classification of ϕMAM1 in the family
Myoviridae
. Bacteriophage ϕMAM1 is virulent, uses capsular polysaccharides as a receptor, and can transduce chromosomal markers at frequencies of up to 7 × 10
−6
transductants per PFU. We also demonstrated transduction of the complete 77-kb oocydin A gene cluster and heterogeneric transduction of a plasmid carrying a type III toxin-antitoxin system. These results support the notion of the potential ecological importance of transducing phages in the acquisition of genes by horizontal gene transfer. Phylogenetic analyses grouped ϕMAM1 within the ViI-like bacteriophages, and genomic analyses revealed that the major differences between ϕMAM1 and other ViI-like phages arise in a region encoding the host recognition determinants. Our results predict that the wider genus of ViI-like phages could be efficient transducing phages, and this possibility has obvious implications for the ecology of horizontal gene transfer, bacterial functional genomics, and synthetic biology.
Two Novel Toxin Variants Revealed by Whole-Genome Sequencing of 175 Clostridium botulinum Type E Strains
Authors: Weedmark K. A. , Lambert D. L. , Mabon P. , Hayden K. L. , Urfano C. J. , Leclair D. , Van Domselaar G. , Austin J. W. , Corbett C. R.
Published: August 2014

We sequenced 175
Clostridium botulinum
type E strains isolated from food, clinical, and environmental sources from northern Canada and analyzed their botulinum neurotoxin (
bont
) coding sequences (CDSs). In addition to
bont/E1
and
bont/E3
variant types, neurotoxin sequence analysis identified two novel BoNT type E variants termed E10 and E11. Strains producing type E10 were found along the eastern coastlines of Hudson Bay and the shores of Ungava Bay, while strains producing type E11 were only found in the Koksoak River region of Nunavik. Strains producing BoNT/E3 were widespread throughout northern Canada, with the exception of the coast of eastern Hudson Bay.
Efficient Simultaneous Saccharification and Fermentation of Inulin to 2,3-Butanediol by Thermophilic Bacillus licheniformis ATCC 14580
Authors: Li Lixiang , Chen Chao , Li Kun , Wang Yu , Gao Chao , Ma Cuiqing , Xu Ping
Published: August 2014

2,3-Butanediol (2,3-BD) is an important starting material for the manufacture of bulk chemicals. For efficient and large-scale production of 2,3-BD through fermentation, low-cost substrates are required. One such substrate, inulin, is a polydisperse fructan found in a wide variety of plants. In this study, a levanase with high inulinase activity and high pH and temperature stability was identified in
Bacillus licheniformis
strain ATCC 14580.
B. licheniformis
strain ATCC 14580 was found to efficiently produce 2,3-BD from fructose at 50°C. Then, the levanase was used for simultaneous saccharification and fermentation (SSF) of inulin to 2,3-BD. A fed-batch SSF yielded 103.0 g/liter 2,3-BD in 30 h, with a high productivity of 3.4 g/liter · h. The results suggest that the SSF process developed with the thermophilic
B. licheniformis
strain used might be a promising alternative for efficient 2,3-BD production from the favorable substrate inulin.
Probing of Microbial Biofilm Communities for Coadhesion Partners
Authors: Ruhl Stefan , Eidt Andreas , Melzl Holger , Reischl Udo , Cisar John O.
Published: August 2014

Investigations of interbacterial adhesion in dental plaque development are currently limited by the lack of a convenient assay to screen the multitude of species present in oral biofilms. To overcome this limitation, we developed a solid-phase fluorescence-based screening method to detect and identify coadhesive partner organisms in mixed-species biofilms. The applicability of this method was demonstrated using coaggregating strains of type 2 fimbrial adhesin-bearing actinomyces and receptor polysaccharide (RPS)-bearing streptococci. Specific adhesin/receptor-mediated coadhesion was detected by overlaying bacterial strains immobilized to a nitrocellulose membrane with a suspended, fluorescein-labeled bacterial partner strain. Coadhesion was comparable regardless of which cell type was labeled and which was immobilized. Formaldehyde treatment of bacteria, either in suspension or immobilized on nitrocellulose, abolished actinomyces type 2 fimbrial adhesin but not streptococcal RPS function, thereby providing a simple method for assigning complementary adhesins and glycan receptors to members of a coadhering pair. The method's broader applicability was shown by overlaying colony lifts of dental plaque biofilm cultures with fluorescein-labeled strains of type 2 fimbriated
Actinomyces naeslundii
or RPS-bearing
Streptococcus oralis
. Prominent coadhesion partners included not only streptococci and actinomyces, as expected, but also other bacteria not identified in previous coaggregation studies, such as adhesin- or receptor-bearing strains of
Neisseria pharyngitis
,
Rothia dentocariosa
, and
Kingella oralis
. The ability to comprehensively screen complex microbial communities for coadhesion partners of specific microorganisms opens a new approach in studies of dental plaque and other mixed-species biofilms.
Dynamics of Dual Infection with Campylobacter jejuni Strains in Chickens Reveals Distinct Strain-to-Strain Variation in Infection Ecology
Authors: Chaloner Gemma , Wigley Paul , Humphrey Suzanne , Kemmett Kirsty , Lacharme-Lora Lizeth , Humphrey Tom , Williams Nicola
Published: August 2014

Although multiple genotypes of
Campylobacter jejuni
may be isolated from the same commercial broiler flock, little is known about the infection dynamics of different genotypes within individuals or their colonization sites within the gut. Single experimental infections with
C. jejuni
M1 (sequence type 137, clonal complex 45) and
C. jejuni
13126 (sequence type 21, clonal complex 21) revealed that 13126 colonized the ceca at significantly higher levels. The dissemination and colonization sites of the two
C. jejuni
strains then were examined in an experimental broiler flock. Two 33-day-old broiler chickens were infected with M1 and two with 13126, and 15 birds were left unchallenged. Cloacal swabs were taken postinfection to determine the colonization and shedding of each strain. By 2 days postinfection (dpi), 8/19 birds were shedding M1 whereas none were shedding 13126. At 8 dpi, all birds were shedding both strains. At 18 dpi, liver and cecal levels of each isolate were quantified, while in 10 birds they also were quantified at nine sites throughout the gastrointestinal (GI) tract. 13126 was found throughout the GI tract, while M1 was largely restricted to the ceca and colon. The livers of 7/19 birds were culture positive for 13126 only. These data show that 13126 has a distinctly different infection biology than strain M1. It showed slower colonization of the lower GI tract but was more invasive and able to colonize at a high level throughout the GI tract. The finding that
C. jejuni
strains have markedly different infection ecologies within the chicken has implications for control in the poultry industry and suggests that the contamination risk of edible tissues is dependent on the isolate involved.
Investigation of the Evolutionary Development of the Genus Bifidobacterium by Comparative Genomics
Authors: Lugli Gabriele Andrea , Milani Christian , Turroni Francesca , Duranti Sabrina , Ferrario Chiara , Viappiani Alice , Mancabelli Leonardo , Mangifesta Marta , Taminiau Bernard , Delcenserie Véronique , van Sinderen Douwe , Ventura Marco
Published: August 2014

The
Bifidobacterium
genus currently encompasses 48 recognized taxa, which have been isolated from different ecosystems. However, the current phylogeny of bifidobacteria is hampered by the relative paucity of genotypic data. Here, we reassessed the taxonomy of this bacterial genus using genome-based approaches, which demonstrated that the previous taxonomic view of bifidobacteria contained several inconsistencies. In particular, high levels of genetic relatedness were shown to exist between particular
Bifidobacterium
taxa which would not justify their status as separate species. The results presented are here based on average nucleotide identity analysis involving the genome sequences for each type strain of the 48 bifidobacterial taxa, as well as phylogenetic comparative analysis of the predicted core genome of the
Bifidobacterium
genus. The results of this study demonstrate that the availability of complete genome sequences allows the reconstruction of a more robust bifidobacterial phylogeny than that obtained from a single gene-based sequence comparison, thus discouraging the assignment of a new or separate bifidobacterial taxon without such a genome-based validation.
Application of Long-Range and Binding Reverse Transcription-Quantitative PCR To Indicate the Viral Integrities of Noroviruses
Authors: Li Dan , De Keuckelaere Ann , Uyttendaele Mieke
Published: August 2014

This study intends to establish and apply methods evaluating both viral capsid and genome integrities of human noroviruses (NoVs), which thus far remain nonculturable. Murine norovirus 1 (MNV-1) and human NoV GII.4 in phosphate-buffered saline suspensions were treated with heat, UV light, or ethanol and detected by reverse transcription-quantitative PCR (RT-qPCR), long-range RT-qPCR, binding RT-qPCR, and binding long-range RT-qPCR. For MNV-1 heated at 60°C for 2 and 30 min, limited reductions of genomic copies (<0.3-log) were obtained by RT-qPCR and long-range RT-qPCR, while the cell-binding pretreatments obtained higher reductions (>1.89-log reduction after 60°C for 30 min by binding long-range RT-qPCR). The human NoV GII.4 was found to be more heat resistant than MNV-1. For both MNV-1 and human NoV GII.4 after UV treatments of 20 and 200 mJ/cm
2
, no significant difference (
P
> 0.05) was observed between the dose-dependent reductions obtained by the four detection methodologies. Treatment of 70% ethanol for 1 min was shown to be more effective for inactivation of both MNV-1 and human NoV GII.4 than the heat and UV treatments used in this study. Subsequently, eight raspberry and four shellfish samples previously shown to be naturally contaminated with human NoVs by RT-qPCR (GI and GII; thus, 24 RT-qPCR signals) were subjected to comparison by this method. RT-qPCR, long-range RT-qPCR, binding RT-qPCR, and binding long-range RT-qPCR detected 20/24, 14/24, 24/24, and 23/24 positive signals, respectively, indicating the abundant presence of intact NoV particles.
Transcriptomic Responses of Phanerochaete chrysosporium to Oak Acetonic Extracts: Focus on a New Glutathione Transferase
Authors: Thuillier Anne , Chibani Kamel , Belli Gemma , Herrero Enrique , Dumarçay Stéphane , Gérardin Philippe , Kohler Annegret , Deroy Aurélie , Dhalleine Tiphaine , Bchini Raphael , Jacquot Jean-Pierre , Gelhaye Eric , Morel-Rouhier Mélanie
Published: August 2014

The first steps of wood degradation by fungi lead to the release of toxic compounds known as extractives. To better understand how lignolytic fungi cope with the toxicity of these molecules, a transcriptomic analysis of
Phanerochaete chrysosporium
genes was performed in the presence of oak acetonic extracts. It reveals that in complement to the extracellular machinery of degradation, intracellular antioxidant and detoxification systems contribute to the lignolytic capabilities of fungi, presumably by preventing cellular damages and maintaining fungal health. Focusing on these systems, a glutathione transferase (
P. chrysosporium
GTT2.1 [PcGTT2.1]) has been selected for functional characterization. This enzyme, not characterized so far in basidiomycetes, has been classified first as a GTT2 compared to the
Saccharomyces cerevisiae
isoform. However, a deeper analysis shows that the GTT2.1 isoform has evolved functionally to reduce lipid peroxidation by recognizing high-molecular-weight peroxides as substrates. Moreover, the GTT2.1 gene has been lost in some non-wood-decay fungi. This example suggests that the intracellular detoxification system evolved concomitantly with the extracellular ligninolytic machinery in relation to the capacity of fungi to degrade wood.
Discovery of Multiple Modified F
430
Coenzymes in Methanogens and Anaerobic Methanotrophic Archaea Suggests Possible New Roles for F
430
in Nature
Authors: Allen Kylie D. , Wegener Gunter , White Robert H.
Published: August 2014

Methane is a potent greenhouse gas that is generated and consumed in anaerobic environments through the energy metabolism of methanogens and
an
aerobic
me
thanotrophic archaea (ANME), respectively. Coenzyme F
430
is essential for methanogenesis, and a structural variant of F
430
, 17
2
-methylthio-F
430
(F
430
-2), is found in ANME and is presumably essential for the anaerobic oxidation of methane. Here we use liquid chromatography–high-resolution mass spectrometry to identify several new structural variants of F
430
in the cell extracts of selected methanogens and ANME.
Methanocaldococcus jannaschii
and
Methanococcus maripaludis
contain an F
430
variant (denoted F
430
-3) that has an M
+
of 1,009.2781. This mass increase of 103.9913 over that of F
430
corresponds to C
3
H
4
O
2
S and is consistent with the addition of a 3-mercaptopropionate moiety bound as a thioether followed by a cyclization. The UV absorbance spectrum of F
430
-3 was different from that of F
430
and instead matched that of an F
430
derivative where the 17
3
keto moiety had been reduced. This is the first report of a modified F
430
in methanogens. In a search for F
430
-2 and F
430
-3 in other methanogens and ANME, we have identified a total of nine modified F
430
structures. One of these compounds may be an abiotic oxidative product of F
430
, but the others represent naturally modified versions of F
430
. This work indicates that F
430
-related molecules have additional functions in nature and will inspire further research to determine the biochemical role(s) of these variants and the pathways involved in their biosynthesis.
Collection and Characterization of Samples for Establishment of a Serum Repository for Lyme Disease Diagnostic Test Development and Evaluation
Authors: Molins Claudia R. , Sexton Christopher , Young John W. , Ashton Laura V. , Pappert Ryan , Beard Charles B. , Schriefer Martin E.
Published: August 2014

Serological assays and a two-tiered test algorithm are recommended for laboratory confirmation of Lyme disease. In the United States, the sensitivity of two-tiered testing using commercially available serology-based assays is dependent on the stage of infection and ranges from 30% in the early localized disease stage to near 100% in late-stage disease. Other variables, including subjectivity in reading Western blots, compliance with two-tiered recommendations, use of different first- and second-tier test combinations, and use of different test samples, all contribute to variation in two-tiered test performance. The availability and use of sample sets from well-characterized Lyme disease patients and controls are needed to better assess the performance of existing tests and for development of improved assays. To address this need, the Centers for Disease Control and Prevention and the National Institutes of Health prospectively collected sera from patients at all stages of Lyme disease, as well as healthy donors and patients with look-alike diseases. Patients and healthy controls were recruited using strict inclusion and exclusion criteria. Samples from all included patients were retrospectively characterized by two-tiered testing. The results from two-tiered testing corroborated the need for novel and improved diagnostics, particularly for laboratory diagnosis of earlier stages of infection. Furthermore, the two-tiered results provide a baseline with samples from well-characterized patients that can be used in comparing the sensitivity and specificity of novel diagnostics. Panels of sera and accompanying clinical and laboratory testing results are now available to Lyme disease serological test users and researchers developing novel tests.
Type I Interferon Protects Mice from Fatal Neurotropic Infection with Langat Virus by Systemic and Local Antiviral Responses
Authors: Weber Elvira , Finsterbusch Katja , Lindquist Richard , Nair Sharmila , Lienenklaus Stefan , Gekara Nelson O , Janik Dirk , Weiss Siegfried , Kalinke Ulrich , Överby Anna K. , Kröger Andrea
Published: August 2014

Vector-borne flaviviruses, such as tick-borne encephalitis virus (TBEV), West Nile virus, and dengue virus, cause millions of infections in humans. TBEV causes a broad range of pathological symptoms, ranging from meningitis to severe encephalitis or even hemorrhagic fever, with high mortality. Despite the availability of an effective vaccine, the incidence of TBEV infections is increasing. Not much is known about the role of the innate immune system in the control of TBEV infections. Here, we show that the type I interferon (IFN) system is essential for protection against TBEV and Langat virus (LGTV) in mice. In the absence of a functional IFN system, mice rapidly develop neurological symptoms and succumb to LGTV and TBEV infections. Type I IFN system deficiency results in severe neuroinflammation in LGTV-infected mice, characterized by breakdown of the blood-brain barrier and infiltration of macrophages into the central nervous system (CNS). Using mice with tissue-specific IFN receptor deletions, we show that coordinated activation of the type I IFN system in peripheral tissues as well as in the CNS is indispensable for viral control and protection against virus induced inflammation and fatal encephalitis.

IMPORTANCE
The type I interferon (IFN) system is important to control viral infections; however, the interactions between tick-borne encephalitis virus (TBEV) and the type I IFN system are poorly characterized. TBEV causes severe infections in humans that are characterized by fever and debilitating encephalitis, which can progress to chronic illness or death. No treatment options are available. An improved understanding of antiviral innate immune responses is pivotal for the development of effective therapeutics. We show that type I IFN, an effector molecule of the innate immune system, is responsible for the extended survival of TBEV and Langat virus (LGTV), an attenuated member of the TBE serogroup. IFN production and signaling appeared to be essential in two different phases during infection. The first phase is in the periphery, by reducing systemic LGTV replication and spreading into the central nervous system (CNS). In the second phase, the local IFN response in the CNS prevents virus-induced inflammation and the development of encephalitis.