bluuebunny/crossref_metadata_2025 huggingface dataset:
When Gray is Good: Gray Markets and Market‐Creating Investments
Authors: Autrey Romana L. , Bova Francesco , Soberman David A.
Published: June 2014

Gray markets arise when an intermediary buys a product in a lower‐priced, often emerging market and resells it to compete with the product's original manufacturer in a higher priced, more developed market. Evidence suggests that gray markets make the original manufacturer worse off globally by eroding profit margins in developed markets. Thus, it is interesting that many firms do not implement control systems to curb gray market activity. Our analysis suggests that one possible explanation lies at the intersection of two economic phenomena: firms investing to build emerging market demand, and investments conferring positive externalities (spillovers) on a rival's demand. We find that gray markets amplify the incentives to invest in emerging markets, because investments increase both emerging market consumption and the gray market's cost base. Moreover, when market‐creating investments confer positive spillovers, each firm builds its own market more efficiently. Thus, firms can be better off with gray markets when investments confer spillovers, provided the spillover effect is sufficiently large. These results provide a perspective on why firms might not implement control systems to prevent gray market distribution in sectors where investment spillovers are common (e.g., the technology sector) and, more broadly, why gray markets persist in the economy.
LAMP assay and rapid sample preparation method for on‐site detection of flavescence dorée phytoplasma in grapevine
Authors: Kogovšek P. , Hodgetts J. , Hall J. , Prezelj N. , Nikolić P. , Mehle N. , Lenarčič R. , Rotter A. , Dickinson M. , Boonham N. , Dermastia M. , Ravnikar M.
Published: June 2014

In Europe the most devastating phytoplasma associated with grapevine yellows (GY) diseases is a quarantine pest, flavescence dorée (FDp), from the 16SrV taxonomic group. The on‐site detection of FDp with an affordable device would contribute to faster and more efficient decisions on the control measures for FDp. Therefore, a real‐time isothermal LAMP assay for detection of FDp was validated according to the EPPO standards and MIQE guidelines. The LAMP assay was shown to be specific and extremely sensitive, because it detected FDp in all leaf samples that were determined to be FDp infected using quantitative real‐time PCR. The whole procedure of sample preparation and testing was designed and optimized for on‐site detection and can be completed in one hour. The homogenization procedure of the grapevine samples (leaf vein, flower or berry) was optimized to allow direct testing of crude homogenates with the LAMP assay, without the need for DNA extraction, and was shown to be extremely sensitive.
Pyramiding Ty‐2 and Ty‐3 genes for resistance to monopartite and bipartite tomato leaf curl viruses of India
Authors: Prasanna H. C. , Sinha D. P. , Rai G. K. , Krishna R. , Kashyap S. P. , Singh N. K. , Singh M. , Malathi V. G.
Published: June 2014

Breeding resistance to whitefly‐transmitted begomoviruses is an important goal of tomato breeding programmes worldwide. So far, resistance to begomoviruses in tomato has been achieved using wild species, and at least five resistance genes (Ty genes) have been studied. The present study was undertaken to combine Ty‐2 and Ty‐3 and to determine the effect of pyramiding on infection of tomato by three diverse begomovirus species. The diagnostic ability of the markers linked to Ty genes was assessed and marker‐assisted selection was used to develop pyramided tomato lines from the crosses between Ty stocks. Five stable pyramided tomato lines that differ in fruit morphology and yield potential were developed. The horticultural performance of pyramided lines in field trials showed that the yield and horticultural traits are well maintained in the plants. The response of these lines was assessed using agroinoculation and field tests in a disease hotspot. The pyramided lines and Ty‐3‐carrying lines exhibited a high level of resistance to the monopartite and two bipartite begomoviruses tested. The pyramided tomato lines developed in this study could be important genetic resources for sustainable tomato production in areas affected by tomato leaf curl virus disease. The combined results of disease resistance tests also showed that Ty‐3 is critical for achieving broad‐spectrum resistance. The limitations of relying on a single gene and the importance of pyramiding are discussed in the light of available evidence on frequent recombination in begomoviruses.
Rb–Sr Dating of Gold‐bearing Pyrites from Wulaga Gold Deposit and its Geological Significance
Authors: Wang Yongbin , Zeng Qingdong , Liu Jianming
Published: June 2014

Wulaga epithermal gold deposit is located in northeast China. Gold mineralization mainly occurs within the crypto‐explosive breccia belt of subvolcanic intrusion. Constraints on the precise timing of mineralization are of fundamental importance for understanding the ore genesis of the Wulaga gold deposit and its mineralization potential. Three hydrothermal stages have been identified: the early veiny quartz–euhedral pyrite stage; the fine pyrite–marcasite–gray or black chalcedony stage; and the late carbonate–pyrite stage. The Rb–Sr dating of gold‐bearing pyrites from the fine pyrite–marcasite–gray or black chalcedony stage is 113.8 ± 4.4 Ma with an initial 87Sr/86Sr ratio of 0.706346 ± 0.000019. The age of the gold deposit is consistent with the age of ore‐bearing volcanic (109–113 Ma) and subvolcanic intrusion (103–112 Ma) within the error limits, and the pyrite initial ratio has an identical value of 87Sr/86Sr to subvolcanic intrusion (0.705547 ± 0.000012). These indicate that crystallization of the wall rock and epithermal gold mineralization was coeval and likely cogenetic. Moreover, a lot of epithermal gold deposits that formed in Early Cretaceous volcanic and subvolcanic intrusions have been discovered in recent years in Heilongjiang province. Combined with the studies of tectonic and magmatic activities, we propose that the formation of the Wulaga gold deposit might be caused by the heated circum‐flow water related to the volcanic–subvolcanic intrusive hydrothermal event triggered by the ancient subduction of the Izanagi plate in the Early Cretaceous.
Development and validation of Neonatal Satisfaction Survey – NSS‐13
Authors: Hagen Inger H. , Vadset Tove B. , Barstad Johan , Svindseth Marit F.
Published: June 2014

Aims

The purpose of this study was to develop and validate a survey to investigate parents' satisfaction with neonatal wards in a population of parents of children with a gestation age of ≥24 weeks to 3 months after full‐term birth.

Method

We explored the literature and conducted three focus groups: two with expert health personnel and one with parents. We tested the survey in a parent population (N = 105) and report the different stages in the validation process along with the full survey, the Neonatal Satisfaction Survey – 13 categories (NSS‐13).

Results

We found 13 subcategories in the Neonatal Satisfaction Survey. The subcategories measure parents' satisfaction with neonatal units based on staff, admission, nurses, anxiety, siblings (parents' perceptions of caring for the siblings of the newborn), information, timeout, doctors, facilities, nutrition, preparation for discharge, trust and visitors. Each subcategory showed acceptable internal consistency. The full version of the Neonatal Satisfaction Survey presents 69 items, and each subcategory contains two to eleven items.

Conclusion

The Neonatal Satisfaction Survey seems suitable to measure parents' satisfaction with neonatal units and can be used in full, but it can also measure subcategories. Parents' satisfaction with neonatal units can be used to improve the quality in such wards. We consider this study as the first in a series to validate the NSS‐13. The full survey with subcategories is presented in this paper.
Experimental Investigations on the Effects of Torque Tightening on the Fatigue Strength of Double‐lap Simple Bolted and Hybrid (Bolted/Bonded) Joints
Authors: Esmaeili F. , Zehsaz M. , Chakherlou T. N.
Published: June 2014

In this paper, the effects of bolt torque tightening on the fatigue strength of double‐lap simple bolted and hybrid (bolted/bonded) joints have been studied experimentally. To do so, two types of joints, that is, double‐lap simple and hybrid (bolted/bonded) joints, were studied. For each type of joints, three sets of specimens were prepared and subjected to the tightening torque of 1, 2.5 and 5 Nm, and then, fatigue tests were carried out at different cyclic longitudinal load levels. Experimental tests results showed that the hybrid joints have better fatigue performance in comparison with the simple bolted joints. In addition, the investigation revealed the positive role of tightening torque on the fatigue life of both simple and hybrid joints.
Genomic sequence of the rare HLA‐A*02:95 allele identified by sequence‐based typing and cloning
Authors: Bultitude W. P. , Mayor N. P. , McWhinnie A. J. , Madrigal J. A. , Marsh S. G. E.
Published: June 2014

Genomic sequence of HLA‐A*02:95 identified in an Anthony Nolan volunteer donor.
Genomic full‐length sequence of two HLA‐A*11 alleles, identified by cloning and sequencing
Authors: Xu Y. P. , Gao S. Q.
Published: June 2014

Genomic full length sequences of HLA‐A*11:01:01 and A*11:02:01 were identified by cloning and sequencing from two different Chinese donors.
Multiple cutaneous histiocytomas treated with lomustine in a dog
Authors: Maina Elisa , Colombo Silvia , Stefanello Damiano
Published: June 2014

Background

Histiocytoma is a common benign neoplasm of young dogs. Multiple histiocytomas are rare. Surgical or medical treatment of solitary tumours is not required in the majority of cases because the tumour usually undergoes spontaneous regression. Therapy is required when lesions are persistent, recurrent, ulcerated or in uncomfortable locations.

Hypothesis/Objectives

To describe a case of canine multiple cutaneous histiocytomas treated with lomustine.

Animal

A 5‐year‐old miniature Pinscher dog was presented with multiple, disseminated, alopecic cutaneous nodules, with no associated systemic signs on initial presentation.

Methods

Histopathological examination of skin biopsies and immunocytochemistry of biopsy imprints were performed. Inguinal lymph node, liver, spleen and bone marrow cytological examination and abdominal ultrasound examination were also performed.

Results

The clinical, histopathological and immunocytochemical findings supported a diagnosis of canine multiple cutaneous histiocytomas. Owing to the increasing number and size of the nodules, medical treatment was initiated. Prednisone and ciclosporin resulted in worsening of lesions. Lomustine orally once monthly led to complete resolution followed by relapse. Metabolic disorders such as increased serum alanine transaminase and alkaline phosphatase activities were recorded, and therapy was stopped. Increase in size of the tumours, severe dullness and anorexia led the owner to elect euthanasia.

Conclusions and clinical importance

To the best of the authors' knowledge, this is the first case report of canine multiple cutaneous histiocytomas treated with lomustine. Lomustine is effective in histiocytic diseases, but adverse effects must be considered because they can be severe and life threatening.
Intestinal adaptations in chronic kidney disease and the influence of gastric bypass surgery
Authors: Hatch Marguerite
Published: June 2014

New Findings

What is the topic of this review?

This review primarily focuses on the compensatory adaptations in gastrointestinal oxalate transport that can impact urinary oxalate excretion in the context of hyperoxaluria, a major risk factor in kidney stone disease.

What advances does it highlight?

The intestine, which is generally considered an absorptive organ, contributes to oxalate homeostasis by playing a role in enteric elimination/excretion of oxalate that has beneficial effects, especially when renal function is compromised. Conversely, when enteric elimination of oxalate is promoted, urinary oxalate can be reduced even in hyperoxaluric conditions. This report highlights the physiological signalling between the kidney and the gut and vice versa.

Studies have shown that compensatory adaptations in gastrointestinal oxalate transport can impact the amount of oxalate excreted by the kidney. Hyperoxaluria is a major risk factor in the formation of kidney stones, and oxalate is derived from both the diet and the liver metabolism of glyoxylate. Although the intestine generally absorbs oxalate from dietary sources and can contribute as much as 50% of urinary oxalate, enteric oxalate elimination plays a significant role when renal function is compromised. While the mechanistic basis for these changes in the direction of intestinal oxalate movements in chronic renal failure involves an upregulation of angiotensin II receptors in the large intestine, enteric secretion/excretion of oxalate can also occur by mechanisms that are independent of angiotensin II. Most notably, the commensal bacterium Oxalobacter sp. interacts with the host enterocyte and promotes the movement of oxalate from the blood into the lumen, resulting in the beneficial effect of significantly lowering urinary oxalate excretion. Changes in the passive permeability of the intestine, such as in steatorrhoea and following gastric bypass, also promote oxalate absorption and hyperoxaluria. In summary, this report highlights the two‐way physiological signalling between the gut and the kidney, which may help to alleviate the consequences of certain kidney diseases.
Neuronal activation by mucosal biopsy supernatants from irritable bowel syndrome patients is linked to visceral sensitivity
Authors: Buhner Sabine , Braak Breg , Li Qin , Kugler Eva Maria , Klooker Tamira , Wouters Mira , Donovan Jemma , Vignali Sheila , Mazzuoli‐Weber Gemma , Grundy David , Boeckxstaens Guy , Schemann Michael
Published: June 2014

New Findings

What is the central question of this study?

Supernatants from colonic mucosal biopsies from patients with irritable bowel syndrome (IBS) activate enteric and dorsal root ganglion (DRG) neurons. Based on the discomfort/pain threshold during rectal distension, IBS patients may be subtyped as normo‐ or hypersensitive. However, the link between neuronal activation and visceral sensitivity remains unknown.

What is the main finding and its importance?

We found that supernatants from hypersensitive IBS patients caused stronger activation of enteric and DRG neurons than supernatants from normosensitive IBS patients. The level of activation correlated with the individual discomfort/pain threshold pressure values. We therefore conclude that mucosal biopsy supernatants have biomarker potential and may, in the future, help to personalize treatment of IBS patients with different visceral sensitivities.

Based on the discomfort/pain threshold during rectal distension, irritable bowel syndrome (IBS) patients may be subtyped as normo‐ or hypersensitive. We previously showed that mucosal biopsy supernatants from IBS patients activated enteric and visceral afferent neurons. We tested the hypothesis that visceral sensitivity is linked to the degree of neuronal activation. Normo‐ and hypersensitive IBS patients were distinguished by their discomfort/pain threshold to rectal balloon distension with a barostat. Using potentiometric and Ca2+ dye imaging, we recorded the response of guinea‐pig enteric submucous and mouse dorsal root ganglion (DRG) neurons, respectively, to mucosal biopsy supernatants from normosensitive (n = 12 tested in enteric neurons, n = 9 tested in DRG) and hypersensitive IBS patients (n = 9, tested in both types of neurons). In addition, we analysed the association between neuronal activation and individual discomfort/pain pressure thresholds. The IBS supernatants evoked Ca2+ transients in DRG neurons and spike discharge in submucous neurons. Submucous and DRG neurons showed significantly stronger responses to supernatants from hypersensitive IBS patients as reflected by higher spike frequency or stronger [Ca2+]i transients in a larger proportion of neurons. The neuroindex as a product of spike frequency or [Ca2+]i transients and proportion of responding neurons correlated significantly with the individual discomfort/pain thresholds of the IBS patients. Supernatants from hypersensitive IBS patients caused stronger activation of enteric and DRG neurons. The level of activation correlated with the individual discomfort/pain threshold pressure values. These findings support our hypothesis that visceral sensitivity is linked to activation of peripheral neurons by biopsy supernatants.
Spatial organization and coordination of slow waves in the mouse anorectum
Authors: Hall K. A. , Ward S. M. , Cobine C. A. , Keef K. D.
Published: June 2014

Key points

The internal anal sphincter (IAS) develops tone important for maintaining high anal pressure and continence whereas tone in the rectum is less.

To investigate tone generation, the electrical properties [membrane potential (Em) and slow waves (SWs)] and morphology of the mouse IAS and distal rectum were compared.

SWs were greatest in amplitude and frequency at the distal end of the IAS and declined toward the rectum. SWs were also coordinated to a greater degree in the circumferential than the oral direction.

The circular muscle was divided into ‘minibundles’ in the IAS but not rectum. Intramuscular interstitial cells of Cajal and platelet‐derived growth factor receptor alpha‐positive cells were present in each minibundle making each a possible candidate for SW generation.

The features that distinguish the IAS from rectum (i.e. depolarized Em, larger and higher frequency SWs and multiunit configuration) are all properties that are predicted to result in greater tone generation.

The internal anal sphincter (IAS) develops tone and is important for maintaining a high anal pressure while tone in the rectum is less. The mechanisms responsible for tone generation in the IAS are still uncertain. The present study addressed this question by comparing the electrical properties and morphology of the mouse IAS and distal rectum. The amplitude of tone and the frequency of phasic contractions was greater in the IAS than in rectum while membrane potential (Em) was less negative in the IAS than in rectum. Slow waves (SWs) were of greatest amplitude and frequency at the distal end of the IAS, declining in the oral direction. Dual microelectrode recordings revealed that SWs were coordinated over a much greater distance in the circumferential direction than in the oral direction. The circular muscle layer of the IAS was divided into five to eight ‘minibundles’ separated by connective tissue septa whereas few septa were present in the rectum. The limited coordination of SWs in the oral direction suggests that the activity in adjacent minibundles is not coordinated. Intramuscular interstitial cells of Cajal and platelet‐derived growth factor receptor alpha‐positive cells were present in each minibundle suggesting a role for one or both of these cells in SW generation. In summary, three important properties distinguish the IAS from the distal rectum: (1) a more depolarized Em; (2) larger and higher frequency SWs; and (3) the multiunit configuration of the muscle. All of these characteristics may contribute to greater tone generation in the IAS than in the distal rectum.
Assessment of in vivo skeletal muscle mitochondrial respiratory capacity in humans by near‐infrared spectroscopy: a comparison with in situ measurements
Authors: Ryan Terence E. , Brophy Patricia , Lin Chien‐Te , Hickner Robert C. , Neufer P. Darrell
Published: June 2014

Key points

In vivo skeletal muscle mitochondrial respiratory capacity was determined from the post‐exercise recovery kinetics of muscle oxygen consumption () measured using near‐infrared spectroscopy (NIRS) in humans.

NIRS recovery rates were compared with the in situ gold standard of high‐resolution respirometry measured in permeabilized muscle fibre bundles prepared from muscle biopsies taken from the same participants.

NIRS‐measured recovery kinetics of were well correlated with maximal ADP‐stimulated mitochondrial respiration in permeabilized fibre bundles.

NIRS provides a cost‐effective, non‐invasive means of assessing in vivo mitochondrial respiratory capacity.

The present study aimed to compare in vivo measurements of skeletal muscle mitochondrial respiratory capacity made using near‐infrared spectroscopy (NIRS) with the current gold standard, namely in situ measurements of high‐resolution respirometry performed in permeabilized muscle fibres prepared from muscle biopsies. Mitochondrial respiratory capacity was determined in 21 healthy adults in vivo using NIRS to measure the recovery kinetics of muscle oxygen consumption following a ∼15 s isometric contraction of the vastus lateralis muscle. Maximal ADP‐stimulated (State 3) respiration was measured in permeabilized muscle fibres using high‐resolution respirometry with sequential titrations of saturating concentrations of metabolic substrates. Overall, the in vivo and in situ measurements were strongly correlated (Pearson's r = 0.61–0.74, all P < 0.01). Bland–Altman plots also showed good agreement with no indication of bias. The results indicate that in vivo NIRS corresponds well with the current gold standard, in situ high‐resolution respirometry, for assessing mitochondrial respiratory capacity.
Mitochondrial reactive oxygen species production and respiratory complex activity in rats with pressure overload‐induced heart failure
Authors: Schwarzer Michael , Osterholt Moritz , Lunkenbein Anne , Schrepper Andrea , Amorim Paulo , Doenst Torsten
Published: June 2014

Key points

Pressure overload induces cardiac hypertrophy developing into heart failure.

During pressure overload‐induced heart failure development in the rat, mitochondrial capacity to produce reactive oxygen species (ROS) increased significantly with the onset of diastolic functional changes.

Treatment to reduce ROS production was able to diminish mitochondrial ROS production but was not able to prevent or delay heart failure development.

The results question a primary role of ROS in the mechanism causing contractile dysfunction under pressure overload.

We investigated the impact of cardiac reactive oxygen species (ROS) during the development of pressure overload‐induced heart failure. We used our previously described rat model where transverse aortic constriction (TAC) induces compensated hypertrophy after 2 weeks, heart failure with preserved ejection fraction at 6 and 10 weeks, and heart failure with systolic dysfunction after 20 weeks. We measured mitochondrial ROS production rates, ROS damage and assessed the therapeutic potential of in vivo antioxidant therapies. In compensated hypertrophy (2 weeks of TAC) ROS production rates were normal at both mitochondrial ROS production sites (complexes I and III). Complex I ROS production rates increased with the appearance of diastolic dysfunction (6 weeks of TAC) and remained high thereafter. Surprisingly, maximal ROS production at complex III peaked at 6 weeks of pressure overload. Mitochondrial respiratory capacity (state 3 respiration) was elevated 2 and 6 weeks after TAC, decreased after this point and was significantly impaired at 20 weeks, when contractile function was also impaired and ROS damage was found with increased hydroxynonenal. Treatment with the ROS scavenger α‐phenyl‐N‐tert‐butyl nitrone or the uncoupling agent dinitrophenol significantly reduced ROS production rates at 6 weeks. Despite the decline in ROS production capacity, no differences in contractile function between treated and untreated animals were observed. Increased ROS production occurs early in the development of heart failure with a peak at the onset of diastolic dysfunction. However, ROS production may not be related to the onset of contractile dysfunction.
Hypometabolism and hypothermia in the rat model of endotoxic shock: independence of circulatory hypoxia
Authors: Corrigan Joshua J. , Fonseca Monique T. , Flatow Elizabeth A. , Lewis Kevin , Steiner Alexandre A.
Published: June 2014

Key points

The hypometabolic, hypothermic response that often replaces fever in endotoxic shock might be consequential to hypoxia, but the available evidence is circumstantial.

Here, this hypothesis was tested in an unprecedented experimental preparation that provides simultaneous measurements of oxygen delivery and consumption in rats whose ability to regulate body temperature has not been disrupted by anaesthetics.

The results are striking for indicating that hypometabolism and hypothermia in endotoxic shock occur independently of global or tissue hypoxia.

The results also demonstrate that a switch in thermal response from fever to hypothermia serves as a pre‐emptive strategy to avoid hypoxia in endotoxic shock.

These findings are potentially relevant to critical care, as interventions aimed at elevating oxygen delivery in patients with septic shock are based on the (perhaps erroneous) premise that circulatory hypoxia is the cause of hypometabolism in this patient population.

We tested the hypothesis that development of hypothermia instead of fever in endotoxic shock is consequential to hypoxia. Endotoxic shock was induced by bacterial lipopolysaccharide (LPS, 500 μg kg−1 i.v.) in rats at an ambient temperature of 22°C. A β3‐adrenergic agonist known to activate metabolic heat production, CL316,243, was employed to evaluate whether thermogenic capacity could be impaired by the fall in oxygen delivery () during endotoxic shock. This possibility was rejected as CL316,243 (0.15 mg kg−1 i.v.) evoked similar rises in oxygen consumption () in the presence and absence of endotoxic shock. Next, to investigate whether a less severe form of circulatory hypoxia could be triggering hypothermia, the circulating volume of LPS‐injected rats was expanded using 6% hetastarch with the intention of improving tissue perfusion and alleviating hypoxia. This intervention attenuated not only the fall in arterial pressure induced by LPS, but also the associated falls in and body temperature. These effects, however, occurred independently of hypoxia, as they were not accompanied by any detectable changes in NAD+/NADH ratios. Further experimentation revealed that even the earliest drops in cardiac output and during endotoxic shock did not precede the reduction in that brings about hypothermia. In fact, and fell in such a synchrony that the / ratio remained unaffected. Only when hypothermia was prevented by exposure to a warm environment (30°C) did an imbalance in the / ratio become evident, and such an imbalance was associated with reductions in the renal and hypothalamic NAD+/NADH ratios. In conclusion, hypometabolism and hypothermia in endotoxic shock are not consequential to hypoxia but serve as a pre‐emptive strategy to avoid hypoxia in this model.
Significant Head Accelerations Can Influence Immediate Neurological Impairments in a Murine Model of Blast-Induced Traumatic Brain Injury
Authors: Gullotti David M. , Beamer Matthew , Panzer Matthew B. , Chia Chen Yung , Patel Tapan P. , Yu Allen , Jaumard Nicolas , Winkelstein Beth , Bass Cameron R. , Morrison Barclay , Meaney David F.
Published: June 2014

Although blast-induced traumatic brain injury (bTBI) is well recognized for its significance in the military population, the unique mechanisms of primary bTBI remain undefined. Animate models of primary bTBI are critical for determining these potentially unique mechanisms, but the biomechanical characteristics of many bTBI models are poorly understood. In this study, we examine some common shock tube configurations used to study blast-induced brain injury in the laboratory and define the optimal configuration to minimize the effect of torso overpressure and blast-induced head accelerations. Pressure transducers indicated that a customized animal holder successfully reduced peak torso overpressures to safe levels across all tested configurations. However, high speed video imaging acquired during the blast showed significant head accelerations occurred when animals were oriented perpendicular to the shock tube axis. These findings of complex head motions during blast are similar to previous reports [Goldstein et al., 2012, “Chronic Traumatic Encephalopathy in Blast-Exposed Military Veterans and a Blast Neurotrauma Mouse Model,” Sci. Transl. Med., 4(134), 134ra160; Sundaramurthy et al., 2012, “Blast-Induced Biomechanical Loading of the Rat: An Experimental and Anatomically Accurate Computational Blast Injury Model,” J. Neurotrauma, 29(13), pp. 2352–2364; Svetlov et al., 2010, “Morphologic and Biochemical Characterization of Brain Injury in a Model of Controlled Blast Overpressure Exposure,” J. Trauma, 69(4), pp. 795–804]. Under the same blast input conditions, minimizing head acceleration led to a corresponding elimination of righting time deficits. However, we could still achieve righting time deficits under minimal acceleration conditions by significantly increasing the peak blast overpressure. Together, these data show the importance of characterizing the effect of blast overpressure on head kinematics, with the goal of producing models focused on understanding the effects of blast overpressure on the brain without the complicating factor of superimposed head accelerations.
Boys Versus Girls: T Lymphocytes in Hypertension
Authors: Price Elvin
Published: June 2014

Immune cells contribute to the development of hypertension in males but not females.
A New Biological Nano-Superglue
Authors: White Nathan J.
Published: June 2014

A fast, easily applied nano-superglue can close both deep and superficial wounds and can attach materials to biological tissues.
The pH-Dependent Expression of the Urease Operon in Streptococcus salivarius Is Mediated by CodY
Authors: Huang Szu-Chuan , Burne Robert A. , Chen Yi-Ywan M.
Published: June 2014

Urease gene expression in
Streptococcus salivarius
57.I, a strain of one of the major alkali producers in the mouth, is induced by acidic pH and excess amounts of carbohydrate. Expression is controlled primarily at the transcriptional level from a promoter, p

ureI

. Recent sequencing analysis revealed a CodY box located 2 bases 5′ to the −35 element of p

ureI

. Using continuous chemostat culture, transcription from p

ureI

was shown to be repressed by CodY, and at pH 7 the repression was more pronounced than that in cells grown at pH 5.5 under both 20 and 100 mM glucose. The direct binding of CodY to p

ureI

was demonstrated by electrophoretic mobility shift assay and chromatin immunoprecipitation (ChIP)–quantitative real-time PCR (qPCR). The result of ChIP-qPCR also confirmed that the regulation of CodY is indeed modulated by pH and the binding of CodY at neutral pH is further enhanced by a limited supply of glucose (20 mM). In the absence of CodY, the C-terminal domain of the RNA polymerase (RNAP) α subunit interacted with the AT tracks within the CodY box, indicating that CodY and RNAP compete for the same binding region. Such regulation could ensure optimal urease expression when the enzyme is most required, i.e., at an acidic growth pH with an excess amount of carbon nutrients.
Trichoderma atroviride Transcriptional Regulator Xyr1 Supports the Induction of Systemic Resistance in Plants
Authors: Reithner Barbara , Mach-Aigner Astrid R. , Herrera-Estrella Alfredo , Mach Robert L.
Published: June 2014

As a result of a transcriptome-wide analysis of the ascomycete
Trichoderma atroviride
, mycoparasitism-related genes were identified; of these, 13 genes were further investigated for differential expression.
In silico
analysis of the upstream regulatory regions of these genes pointed to xylanase regulator 1 (Xyr1) as a putatively involved regulatory protein. Transcript analysis of the
xyr1
gene of
T. atroviride
in confrontation with other fungi allowed us to determine that
xyr1
levels increased during mycoparasitism. To gain knowledge about the precise role of Xyr1 in the mycoparasitic process, the corresponding gene was deleted from the
T. atroviride
genome. This resulted in strong reductions in the transcript levels of
axe1
and
swo1
, which encode accessory cell wall-degrading enzymes considered relevant for mycoparasitism. We also analyzed the role of Xyr1 in the
Trichoderma
-
Arabidopsis
interaction, finding that the plant response elicited by
T. atroviride
is delayed if Xyr1 is missing in the fungus.
Abundance and Diversity of Soybean-Nodulating Rhizobia in Black Soil Are Impacted by Land Use and Crop Management
Authors: Yan Jun , Han Xiao Zeng , Ji Zhao Jun , Li Yan , Wang En Tao , Xie Zhi Hong , Chen Wen Feng
Published: June 2014

To investigate the effects of land use and crop management on soybean rhizobial communities, 280 nodule isolates were trapped from 7 fields with different land use and culture histories. Besides the known
Bradyrhizobium japonicum
, three novel genospecies were isolated from these fields. Grassland (GL) maintained a higher diversity of soybean bradyrhizobia than the other cultivation systems. Two genospecies (
Bradyrhizobium
spp. I and III) were distributed widely in all treatments, while
Bradyrhizobium
sp. II was found only in GL treatment. Cultivation with soybeans increased the rhizobial abundance and diversity, except for the soybean monoculture (S-S) treatment. In monoculture systems, soybeans favored
Bradyrhizobium
sp. I, while maize and wheat favored
Bradyrhizobium
sp. III. Fertilization decreased the rhizobial diversity indexes but did not change the species composition. The organic carbon (OC) and available phosphorus (AP) contents and pH were the main soil parameters positively correlated with the distribution of
Bradyrhizobium
spp. I and II and
Bradyrhizobium japonicum
and negatively correlated with
Bradyrhizobium
sp. III. These results revealed that different land uses and crop management could not only alter the diversity and abundance of soybean rhizobia, but also change interactions between rhizobia and legume or nonlegume plants, which offered novel information about the biogeography of rhizobia.
Optimization of Multilocus Sequence Analysis for Identification of Species in the Genus Vibrio
Authors: Gabriel Michael W. , Matsui George Y. , Friedman Robert , Lovell Charles R.
Published: June 2014

Multilocus sequence analysis (MLSA) is an important method for identification of taxa that are not well differentiated by 16S rRNA gene sequences alone. In this procedure, concatenated sequences of selected genes are constructed and then analyzed. The effects that the number and the order of genes used in MLSA have on reconstruction of phylogenetic relationships were examined. The
recA
,
rpoA
,
gapA
, 16S rRNA gene,
gyrB
, and
ftsZ
sequences from 56 species of the genus
Vibrio
were used to construct molecular phylogenies, and these were evaluated individually and using various gene combinations. Phylogenies from two-gene sequences employing
recA
and
rpoA
in both possible gene orders were different. The addition of the
gapA
gene sequence, producing all six possible concatenated sequences, reduced the differences in phylogenies to degrees of statistical (bootstrap) support for some nodes. The overall statistical support for the phylogenetic tree, assayed on the basis of a reliability score (calculated from the number of nodes having bootstrap values of ≥80 divided by the total number of nodes) increased with increasing numbers of genes used, up to a maximum of four. No further improvement was observed from addition of the fifth gene sequence (
ftsZ
), and addition of the sixth gene (
gyrB
) resulted in lower proportions of strongly supported nodes. Reductions in the numbers of strongly supported nodes were also observed when maximum parsimony was employed for tree construction. Use of a small number of gene sequences in MLSA resulted in accurate identification of
Vibrio
species.
Microbial Community Degradation of Widely Used Quaternary Ammonium Disinfectants
Authors: Oh Seungdae , Kurt Zohre , Tsementzi Despina , Weigand Michael R. , Kim Minjae , Hatt Janet K. , Tandukar Madan , Pavlostathis Spyros G. , Spain Jim C. , Konstantinidis Konstantinos T.
Published: June 2014

Benzalkonium chlorides (BACs) are disinfectants widely used in a variety of clinical and environmental settings to prevent microbial infections, and they are frequently detected in nontarget environments, such as aquatic and engineered biological systems, even at toxic levels. Therefore, microbial degradation of BACs has important ramifications for alleviating disinfectant toxicity in nontarget environments as well as compromising disinfectant efficacy in target environments. However, how natural microbial communities respond to BAC exposure and what genes underlie BAC biodegradation remain elusive. Our previous metagenomic analysis of a river sediment microbial community revealed that BAC exposure selected for a low-diversity community, dominated by several members of the
Pseudomonas
genus that quickly degraded BACs. To elucidate the genetic determinants of BAC degradation, we conducted time-series metatranscriptomic analysis of this microbial community during a complete feeding cycle with BACs as the sole carbon and energy source under aerobic conditions. Metatranscriptomic profiles revealed a candidate gene for BAC dealkylation, the first step in BAC biodegradation that results in a product 500 times less toxic. Subsequent biochemical assays and isolate characterization verified that the putative amine oxidase gene product was functionally capable of initiating BAC degradation. Our analysis also revealed cooperative interactions among community members to alleviate BAC toxicity, such as the further degradation of BAC dealkylation by-products by organisms not encoding amine oxidase. Collectively, our results advance the understanding of BAC aerobic biodegradation and provide genetic biomarkers to assess the critical first step of this process in nontarget environments.
Transcriptional Regulator LsrB of Sinorhizobium meliloti Positively Regulates the Expression of Genes Involved in Lipopolysaccharide Biosynthesis
Authors: Tang Guirong , Wang Ying , Luo Li
Published: June 2014

Rhizobia induce nitrogen-fixing nodules on host legumes, which is important in agriculture and ecology. Lipopolysaccharide (LPS) produced by rhizobia is required for infection or bacteroid survival in host cells. Genes required for LPS biosynthesis have been identified in several
Rhizobium
species. However, the regulation of their expression is not well understood. Here,
Sinorhizobium meliloti
LsrB, a member of the LysR family of transcriptional regulators, was found to be involved in LPS biosynthesis by positively regulating the expression of the
lrp3-lpsCDE
operon. An
lsrB
in-frame deletion mutant displayed growth deficiency, sensitivity to the detergent sodium dodecyl sulfate, and acidic pH compared to the parent strain. This mutant produced slightly less LPS due to lower expression of the
lrp3
operon. Analysis of the transcriptional start sites of the
lrp3
and
lpsCDE
gene suggested that they constitute one operon. The expression of
lsrB
was positively autoregulated. The promoter region of
lrp3
was specifically precipitated by anti-LsrB antibodies
in vivo
. The promoter DNA fragment containing TN11A motifs was bound by the purified LsrB protein
in vitro
. These new findings suggest that
S. meliloti
LsrB is associated with LPS biosynthesis, which is required for symbiotic nitrogen fixation on some ecotypes of alfalfa plants.
F-Box Protein RcyA Controls Turnover of the Kinesin-7 Motor KipA in Aspergillus nidulans
Authors: Herrero Saturnino , Takeshita Norio , Fischer Reinhard
Published: June 2014

Fungal filamentous growth depends on continuous membrane insertion at the tip, the delivery of membrane-bound positional markers, and the secretion of enzymes for cell wall biosynthesis. This is achieved through exocytosis. At the same time, polarized growth requires membrane and protein recycling through endocytosis. Endocytic vesicles are thought to enter the protein degradation pathway or recycle their content to the cell surface. In
Saccharomyces cerevisiae
, the Rcy1 F-box protein is involved in the recycling process of a v-SNARE protein. We identified a Rcy1 orthologue, RcyA, in the filamentous fungus
Aspergillus nidulans
as a protein interacting with the KipA kinesin-7 motor protein in a yeast two-hybrid screen. The interaction was confirmed through bimolecular fluorescence complementation. RcyA possesses an F-box domain at the N terminus and a prenylation (CaaX) motif at the C terminus. RcyA shows also similarity to Sec10, a component of the exocyst complex. The RcyA protein localized to the hyphal tip and forming septa, likely through transportation on secretory vesicles and partially on early endosomes, but independently of KipA. Deletion of
rcyA
did not cause severe morphological changes but caused partial defects in the recycling of the SynA v-SNARE protein and the positioning of the cell end markers TeaA and TeaR. In addition, deletion of
rcyA
led to increased concentrations of the KipA protein, whereas the transcript concentration was unaffected. These results suggest that RcyA probably labels KipA for degradation and thereby controls the protein amount of KipA.
Toxoplasma gondii Development of Its Replicative Niche: in Its Host Cell and Beyond
Authors: Blader Ira J. , Koshy Anita A.
Published: June 2014

Intracellular pathogens can replicate efficiently only after they manipulate and modify their host cells to create an environment conducive to replication. While diverse cellular pathways are targeted by different pathogens, metabolism, membrane and cytoskeletal architecture formation, and cell death are the three primary cellular processes that are modified by infections.Toxoplasma gondiiis an obligate intracellular protozoan that infects ∼30% of the world's population and causes severe and life-threatening disease in developing fetuses, in immune-comprised patients, and in certain otherwise healthy individuals who are primarily found in South America. The high prevalence ofToxoplasmain humans is in large part a result of its ability to modulate these three host cell processes. Here, we highlight recent work defining the mechanisms by whichToxoplasmainteracts with these processes. In addition, we hypothesize why some processes are modified not only in the infected host cell but also in neighboring uninfected cells.
How Does It Kill?: Understanding the Candidacidal Mechanism of Salivary Histatin 5
Authors: Puri Sumant , Edgerton Mira
Published: June 2014

Histatins are salivary cationic peptides that provide the first line of defense against oral candidiasis caused by
Candida albicans
. This minireview presents a critical evaluation of our knowledge of the candidacidal mechanism of histatin 5 (Hst 5). Hst 5 is the most potent among all histatin family members with regard to its antifungal activity. The mode of action of Hst 5 has been a subject of intense debate. Unlike other classical host innate immune proteins, pore formation or membrane lysis by Hst 5 has largely been disproven, and it is now known that all targets of Hst 5 are intracellular. Hst 5 binds
C. albicans
cell wall proteins (Ssa1/2) and glycans and is taken up by the cells through fungal polyamine transporters in an energy-dependent manner. Once inside the fungal cells, Hst 5 may affect mitochondrial functions and cause oxidative stress; however, the ultimate cause of cell death is by volume dysregulation and ion imbalance triggered by osmotic stress. Besides these diverse targets, a novel mechanism based on the metal binding abilities of Hst 5 is discussed. Finally, translational approaches for Hst 5, based on peptide design and synergy with other known drugs, are considered a step forward for bench-to-bed application of Hst 5.
Membrane Fluidity and Temperature Sensing Are Coupled via Circuitry Comprised of Ole1, Rsp5, and Hsf1 in Candida albicans
Authors: Leach Michelle D. , Cowen Leah E.
Published: June 2014

Temperature is a ubiquitous environmental variable which can profoundly influence the physiology of living cells as it changes over time and space. When yeast cells are exposed to a sublethal heat shock, normal metabolic functions become repressed and the heat shock transcription factor Hsf1 is activated, inducing heat shock proteins (HSPs).
Candida albicans
, the most prevalent human fungal pathogen, is an opportunistic pathogen that has evolved as a relatively harmless commensal of healthy individuals. Even though
C. albicans
occupies thermally buffered niches, it has retained the classic heat shock response, activating Hsf1 during slow thermal transitions such as the increases in temperature suffered by febrile patients. However, the mechanism of temperature sensing in fungal pathogens remains enigmatic. A few studies with
Saccharomyces cerevisiae
suggest that thermal stress is transduced into a cellular signal at the level of the membrane. In this study, we manipulated the fluidity of
C. albicans
membrane to dissect mechanisms of temperature sensing. We determined that in response to elevated temperature, levels of
OLE1
, encoding a fatty acid desaturase, decrease. Subsequently, loss of
OLE1
triggers expression of
FAS2
, encoding a fatty acid synthase. Furthermore, depletion of
OLE1
prevents full activation of Hsf1, thereby reducing
HSP
expression in response to heat shock. This reduction in Hsf1 activation is attributable to the E3 ubiquitin ligase Rsp5, which regulates
OLE1
expression. To our knowledge, this is the first study to define a molecular link between fatty acid synthesis and the heat shock response in the fungal kingdom.
Identification of Microorganisms by FilmArray and Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry Prior to Positivity in the Blood Culture System
Authors: Almuhayawi Mohammed , Altun Osman , Strålin Kristoffer , Özenci Volkan
Published: June 2014

In this study, we investigated the performance of the FilmArray and matrix-assisted laser desorption ionization–time of flight mass spectrometry (MALDI-TOF MS) in identifying microorganisms from blood culture (BC) bottles prior to positivity. First, we used simulated BacT/Alert FA Plus BC bottles with five each for
Escherichia coli
and
Staphylococcus aureus
isolates. The FilmArray identified all 10 isolates before BC positivity with 9/10 at 5 h and 1 at 7.5 h after incubation in the BC system. MALDI-TOF MS failed to identify the isolates prior to positivity. When the bottles were incubated for 2.5 h at room temperature (RT) before we put them into the BC system, the FilmArray identified 6/10 at 2.5 h and the remaining 4 at 5 h. Finally, we tested simulated BC bottles after incubation at RT. Interestingly, 9/10 isolates were identified with the FilmArray after 8 h of incubation at RT. Second, we studied clinical BC bottles in quadruplicate. When three-fourths of the parallel bottles signaled positive, the FilmArray was run on the fourth nonsignaled bottle and was found to be positive in 14/15 such cases. Third, we analyzed the performance of the FilmArray in the identification of microorganisms from clinical BC bottles before incubation in the system. Two milliliters of broth from 400 BC bottles was collected after arrival at the laboratory and stored at −70°C. Sixteen bottles later signaled positive in the system. When the frozen broth from these bottles was analyzed, the FilmArray identified all the microorganisms in 8/16 bottles prior to incubation in the BC system. This study shows that the FilmArray can identify microorganisms from BC bottles prior to positivity and in some cases even prior to incubation in the BC system.