bluuebunny/crossref_metadata_2025 huggingface dataset:
Novel Roles of Cytoplasmic ICP0: Proteasome-Independent Functions of the RING Finger Are Required To Block Interferon-Stimulated Gene Production but Not To Promote Viral Replication
Authors: Taylor Kathryne E. , Chew Marianne V. , Ashkar Ali A. , Mossman Karen L.
Published: May 2014

The immediate-early protein ICP0 from herpes simplex virus 1 (HSV-1) plays pleiotropic roles in promoting viral lytic replication and reactivation from latency. Most of the known actions of ICP0 occur in the nucleus and are thought to involve the E3 ubiquitin ligase activity of its RING finger domain, which targets proteins for degradation via the proteasome. Although ICP0 translocates to the cytoplasm as the infection progresses, little is known about its activities in this location. Here, we show that cytoplasmic ICP0 has two distinct functions. In primary cell cultures and in an intravaginal mouse model, cytoplasmic ICP0 promotes viral replication in the absence of an intact RING finger domain. Additionally, ICP0 blocks the activation of interferon regulatory factor 3 (IRF3), a key transcription factor of the innate antiviral response, in a mechanism that requires the RING finger domain but not the proteasome. To our knowledge, this is the first observation of a proteasome-independent function of the RING finger domain of ICP0. Collectively, these results underscore the importance of cytoplasm-localized ICP0 and the diverse nature of its activities.

IMPORTANCE
Despite ICP0 being a well-studied viral protein, the significance of its cytoplasmic localization has been largely overlooked. This is, in part, because common experimental manipulations result in the restriction of ICP0 to the nucleus. By overcoming this constraint, we both further characterize the ability of cytoplasmic ICP0 to inhibit antiviral signaling and show that ICP0 at this site has unexpected activities in promoting viral replication. This demonstrates the importance of considering location when analyzing protein function and adds a new perspective to our understanding of this multifaceted protein.
Replication of Many Human Viruses Is Refractory to Inhibition by Endogenous Cellular MicroRNAs
Authors: Bogerd Hal P. , Skalsky Rebecca L. , Kennedy Edward M. , Furuse Yuki , Whisnant Adam W. , Flores Omar , Schultz Kimberly L. W. , Putnam Nicole , Barrows Nicholas J. , Sherry Barbara , Scholle Frank , Garcia-Blanco Mariano A. , Griffin Diane E. , Cullen Bryan R.
Published: May 2014

The issue of whether viruses are subject to restriction by endogenous microRNAs (miRNAs) and/or by virus-induced small interfering RNAs (siRNAs) in infected human somatic cells has been controversial. Here, we address this question in two ways. First, using deep sequencing, we demonstrate that infection of human cells by the RNA virus dengue virus (DENV) or West Nile virus (WNV) does not result in the production of any virus-derived siRNAs or viral miRNAs. Second, to more globally assess the potential of small regulatory RNAs to inhibit virus replication, we used gene editing to derive human cell lines that lack a functional Dicer enzyme and that therefore are unable to produce miRNAs or siRNAs. Infection of these cells with a wide range of viruses, including DENV, WNV, yellow fever virus, Sindbis virus, Venezuelan equine encephalitis virus, measles virus, influenza A virus, reovirus, vesicular stomatitis virus, human immunodeficiency virus type 1, or herpes simplex virus 1 (HSV-1), failed to reveal any enhancement in the replication of any of these viruses, although HSV-1, which encodes at least eight Dicer-dependent viral miRNAs, did replicate somewhat more slowly in the absence of Dicer. We conclude that most, and perhaps all, human viruses have evolved to be resistant to inhibition by endogenous human miRNAs during productive replication and that dependence on a cellular miRNA, as seen with hepatitis C virus, is rare. How viruses have evolved to avoid inhibition by endogenous cellular miRNAs, which are generally highly conserved during metazoan evolution, remains to be determined.

IMPORTANCE
Eukaryotic cells express a wide range of small regulatory RNAs, including miRNAs, that have the potential to inhibit the expression of mRNAs that show sequence complementarity. Indeed, previous work has suggested that endogenous miRNAs have the potential to inhibit viral gene expression and replication. Here, we demonstrate that the replication of a wide range of pathogenic viruses is not enhanced in human cells engineered to be unable to produce miRNAs, indicating that viruses have evolved to be resistant to inhibition by miRNAs. This result is important, as it implies that manipulation of miRNA levels is not likely to prove useful in inhibiting virus replication. It also focuses attention on the question of how viruses have evolved to resist inhibition by miRNAs and whether virus mutants that have lost this resistance might prove useful, for example, in the development of attenuated virus vaccines.
Rules and Exceptions: Sialic Acid Variants and Their Role in Determining Viral Tropism
Authors: Stehle Thilo , Khan Zaigham M.
Published: May 2014

Sialic acids decorate the surfaces of most mammalian cells and are used by many viruses as attachment receptors. In contrast to other mammals, humans cannot synthesize a version of sialic acid known as
N
-glycolyl neuraminic acid. This difference is exploited by some viruses to establish tropism. Here we compare recently determined structures of closely related animal and human polyomaviruses and examine their strategies for engaging specific sialic acid variants.
Is that lumbar disc symptomatic? Herniated lumbar disc associated with contralateral radiculopathy
Authors: Abdul Jalil Muhammad Fahmi , Lam Miu Fei , Wang Yi Yuen
Published: May 2014

Herniated lumbar disc may be asymptomatic or associated with lower limb radiculopathy. Most spinal surgeons would offer surgery following a period of conservative measures if the radiological and clinical findings correlate. However, the existing dictum that lumbar radiculopathy should correlate with ipsilateral lumbar disc herniation may not be accurate as it can rarely present with contralateral sciatica. Literature regarding this phenomenon is scarce. Therefore, we report a patient with herniated lumbar disc presenting with predominantly contralateral motor weakness radiculopathy, which resolved after discectomy.
Mild meningococcaemia, pyrexia protocols and a problematic public health response
Authors: Allen Luke Nelson
Published: May 2014

Meningococcal septicaemia was masked by an acute tonsillitis in a university student presenting to the emergency department. The diagnosis was made as a result of routine blood culture and resulted in the patient being recalled for appropriate treatment. Although the patient was relatively well, public health contact tracing and prophylaxis ensued as per any other case of meningococcal disease. This case highlights the protean manifestations of meningococcaemia and the importance of evidence-based protocols. Blood culture guidelines led to the diagnosis in the absence of clinical suspicion and in sharp contrast to the paucity of evidence informing the public health response in this area.
An unusual cause of a fall: an unusual presentation of lung carcinoma
Authors: Kowa Xin-Ying , Hyams Catherine , Farrugia Mark , Lightowlers Sara
Published: May 2014

An 80-year-old gentleman presented with a fall and subtle symptoms suggestive of a cerebellar lesion, on a background of hemiparesis due to a previous cerebral vascular accident. On admission it was thought that changes on a chest radiograph were consistent with a community-acquired pneumonia. A CT of the head showed a space-occupying lesion in the right cerebellum with surrounding oedema. A previous MRI of the brain performed 8 months earlier for investigation of dementia showed evidence of this lesion although it was significantly smaller and without midline shift; however, this had not been discussed on the MRI report. A subsequent CT of the thorax confirmed a thick-walled cavitating mass in the left upper lobe, with biopsy of this lesion and bronchial washings showing metaplastic and atypical cells suggestive of adenocarcinoma. This report highlights both the importance of accurate imaging reporting, even of incidental findings, and the insidious nature of lung malignancy and its broad clinical presentation.
Missing the beat: arrhythmia as a presenting feature of eosinophilic granulomatosis with polyangiitis
Authors: Sharpley Faye Amelia
Published: May 2014

Palpitations are a common presenting symptom in patients attending the emergency department; however, eosinophilic infiltration of the myocardium is rarely the cause. This case describes a 77-year-old woman who presents with sudden onset palpitations and is later diagnosed with eosinophilic granulomatosis with polyangiitis (previously known as Churg-Strauss syndrome). Cardiac involvement does occur in 50% of cases but heart failure, myocarditis, pericarditis, constrictive pericarditis and myocardial infarction are much more commonly recognised complications. Arrhythmia is less well described. In this report, we propose eosinophilic inflammation as the precipitant for an aberrant conduction pathway.
An unexpected cause of small bowel obstruction in an adult patient: midgut volvulus
Authors: Söker Gökhan , Yılmaz Cengiz , Karateke Faruk , Gülek Bozkurt
Published: May 2014

The most important complication of intestinal malrotation is midgut volvulus because it may lead to intestinal ischaemia and necrosis. A 29-year-old male patient was admitted to the emergency department with abdominal pain. Ultrasonography (US), colour Doppler ultrasonography (CDUS), CT and barium studies were carried out. On US and CDUS, twisting of intestinal segments around the superior mesenteric artery (SMA) and superior mesenteric vein (SMV) and alteration of the SMA–SMV relationship were detected. CT demonstrated that the small intestine was making a rotation around the SMA and SMV, which amounted to more than 360°. The upper gastrointestinal barium series revealed a corkscrew appearance of the duodenum and proximal jejunum, which is a pathognomonic finding of midgut volvulus. Prior knowledge of characteristic imaging findings of midgut volvulus is essential in order to reach proper diagnosis and establish proper treatment before the development of intestinal ischaemia and necrosis.
Morphology and ontogeny of tannin-producing structures in two tropical legume trees
Authors: de Barros Thais Cury , Teixeira Simone Pádua
Published: May 2014

Two legume trees largely known as tannin producers — Dimorphandra mollis Benth. (Caesalpinioideae) and Stryphnodendron adstringens (Mart.) Coville (Mimosoideae) — were used as models to elucidate the morphology and ontogeny of tannin cells. Vegetative parts of plants were processed for observation using light and electron microscopy (scanning and transmission). Idioblasts, found even in young plants of both species, and secretory trichomes, observed in vegetative buds of mature plants of S. adstringens, are responsible for tannin production. The tanniniferous idioblasts originate from protoderm and also from ground meristem cells. The ground meristem proved to be the best place to study the development of tanniniferous idioblasts at different stages of development, which allowed us to monitor the production and accumulation of tannins in the same tissue. Our data indicate that there is a relationship between the production of tannins and the process of vacuolation of tanniniferous cells. The results also indicate the probable performance of rough endoplasmic reticulum (RER) and plastids in the production of tannins.
Halogen bond interactions enhanced by sodium bonds — Theoretical evidence for cooperative and substitution effects in NCX···NCNa···NCY complexes (X = F, Cl, Br, I; Y = H, F, OH)
Authors: Esrafili Mehdi D. , Mohammadirad Nafiseh
Published: May 2014

Quantum chemical calculations are performed to study cooperativity and substituent effects between halogen bond and sodium bond interactions in NCX···NCNa···NCY complexes, where X = F, Cl, Br, I and Y = H, F, OH. These effects are studied in terms of equilibrium geometry, interaction energy,15N nuclear magnetic resonance (NMR) parameters, and electron density analysis of the complexes at the MP2/aug-cc-pVTZ level. The X···N and Na···N bond lengths in the ternary systems are always shorter than those in the corresponding dyads. In each triad, the decrease in the halogen bond length is far greater than that of the sodium bond. In all triads studied, a favorable cooperativity is found with values that range between –0.30 and –1.72 kcal/mol. It is revealed that NMR parameters at the site of the nitrogen atom of the NCNa molecule can be regarded as a good description to quantify the degree of cooperative effects in the NCX···NCNa···NCY systems. The nature of halogen bond and sodium bond interactions of the complexes is analyzed using parameters derived from the quantum theory of atoms in molecules methodology and energy decomposition analysis.
Longitudinal variation in community structure of floodplain fishes along two rivers of the southeastern USA
Authors: Garnett Jeffrey A. , Batzer Darold P.
Published: May 2014

Many studies have examined whether longitudinal variation exists in fish communities within stream channels. Yet despite the acknowledged importance of adjacent floodplain ecosystems, no study has investigated longitudinal variation in fish communities in floodplain wetlands. We conducted a study to determine if distinct fish communities exist between upper and lower river floodplains. We predicted that floodplains in the upper portions of a watershed will be dominated by opportunistic riverine fish. In contrast, floodplains farther downstream in the watershed will primarily consist of obligate floodplain fishes. We sampled fishes at 21 floodplain sites along the lengths of the Oconee–Altamaha and Broad–Savannah river systems of Georgia, USA, during the 2008 and 2009 flood seasons. A modified mark–recapture method was utilized to estimate probabilities of species detection and occupancy. Ordination and analysis of similarity using species site occupancy estimates identified two unique communities related to floodplain position, and indicator species analysis supported our hypothesis of community distinction based on life history traits. Water resource managers should consider how flood pulse character influences fish communities when regulating flow regimes.
Lake Michigan offshore ecosystem structure and food web changes from 1987 to 2008
Authors: Rogers Mark W. , Bunnell David B. , Madenjian Charles P. , Warner David M.
Published: May 2014

Ecosystems undergo dynamic changes owing to species invasions, fisheries management decisions, landscape modifications, and nutrient inputs. At Lake Michigan, new invaders (e.g., dreissenid mussels (Dreissena spp.), spiny water flea (Bythotrephes longimanus), round goby (Neogobius melanostomus)) have proliferated and altered energy transfer pathways, while nutrient concentrations and stocking rates to support fisheries have changed. We developed an ecosystem model to describe food web structure in 1987 and ran simulations through 2008 to evaluate changes in biomass of functional groups, predator consumption, and effects of recently invading species. Keystone functional groups from 1987 were identified as Mysis, burbot (Lota lota), phytoplankton, alewife (Alosa pseudoharengus), nonpredatory cladocerans, and Chinook salmon (Oncorhynchus tshawytscha). Simulations predicted biomass reductions across all trophic levels and predicted biomasses fit observed trends for most functional groups. The effects of invasive species (e.g., dreissenid grazing) increased across simulation years, but were difficult to disentangle from other changes (e.g., declining offshore nutrient concentrations). In total, our model effectively represented recent changes to the Lake Michigan ecosystem and provides an ecosystem-based tool for exploring future resource management scenarios.
Nonlocal metric realizations of MOND
Authors: Woodard R.P.
Published: May 2014

I discuss relativistic extensions of MOND in which the metric couples normally to matter. I argue that MOND might be a residual effect from the vacuum polarization of infrared gravitons produced during primordial inflation. If so, MOND corrections to the gravitational field equations would be nonlocal. Nonlocality also results when one constructs metric field equations that reproduce the Tully–Fisher relation, along with sufficient weak lensing. I give the full field equations for the simplest class of models, and I specialize these equations to the geometries relevant for cosmology. I conclude by sketching the direction of future studies.
Graphene and high energy physics
Authors: Sidharth B. G.
Published: May 2014

We briefly comment upon the parallel between graphene and high energy fermions and explore the possibility of using the former as a test bed for the latter. We also point out that there are parallels to quantum gravity approaches, which indeed provide a novel explanation for such effects as the Fractional Quantum Hall Effect (FQAE).
The GBAR experiment
Authors: van der Werf D. P. ,   (for the GBAR Collaboration)
Published: May 2014

The classical Weak Equivalence Principle has not yet been tested using antimatter in matter gravitational fields. The GBAR (Gravitational Behaviour of Antihydrogen at Rest) experiment, recently approved by CERN, proposes to measure the free-fall acceleration of antihydrogen. In this experiment, positive antihydrogen ions will be produced, and subsequently cooled down using laser cooled Be + ions. Then, when a temperature of around 20 μK has been reached, the excess positron will be detached and the free-fall time will be measured using the antiproton annihilation products. An overview of the experiment will be given together with its present status.
Pore-pressure cycling experiments on Mx80 Bentonite
Authors: Graham C. C. , Harrington J. F. , Cuss R. J. , Sellin P.
Published: May 2014

The Swedish concept for geological disposal of radioactive waste involves the use of bentonite as part of an engineered barrier system. A primary function of the bentonite is its ability to swell when hydrated by its surroundings. One particular uncertainty is the impact on this function, resulting from deviations in pore-water pressure,
p

w

, from expected
in situ
hydrostatic conditions. We present results from a series of laboratory experiments designed to investigate the form of the relationship between swelling pressure and
p

w

, for compacted Mx80 bentonite, from low to elevated applied water pressure conditions. The experiments were conducted using constant volume cells, designed to allow the total stresses acting on the surrounding vessel to be monitored (at five locations) during clay swelling. The results demonstrate that swelling pressure reduces non-linearly with increasing
p

w

, becoming less sensitive to changes at elevated pressures. After cyclic loading a marked hysteresis was also observed, with swelling pressure remaining elevated after a subsequent reduction in applied water pressure. Such behaviour may impact the mechanical and transport properties of the bentonite and its resulting performance. However, such hysteric behaviour was not always observed. Further testing is required to better understand the causes of this phenomenon and the controls on such behaviour.
Full-scale 3D modelling of a nuclear waste repository in the Callovo-Oxfordian clay. Part 1: thermo-hydraulic two-phase transport of water and hydrogen
Authors: Brommundt J. , Kaempfer Th. U. , Enssle C. P. , Mayer G. , Wendling J.
Published: May 2014

The development of deep geological repositories for nuclear waste requires a sound system understanding, to which performance analyses by means of numerical flow and transport simulations form an important contribution. In this context, we present modelling studies of the thermo-hydraulic two-phase transport of water and hydrogen in the planned French repository in this paper, part 1. Part 2 of this paper uses the same modelling concept extended to include radionuclide transport. The numerical TOUGH2-MP-model encompasses the host rock from cap to bed rock, including the repository in full 3D. We compare two repository layouts and assess their robustness through sensitivity simulations with varied key parameters and hydrogen generation regimes, and a coupling of hydrogen generation to water availability. The results show that a continuous gas phase functions as balancing system between connected waste areas. Its extension and continuity depend on the permeability and diffusion parameters, and the amount and temporal regime of hydrogen production. Naturally, the available storage volumes have an impact as well. The layout with larger storage volume behaves more benevolenty towards parameter variations. Overall, both layouts perform satisfactorily. Our methodology enables detailed studies of the global system behaviour and helps to compare different repository layouts.
Full-scale 3D modelling of a nuclear waste repository in the Callovo-Oxfordian clay. Part 2: thermo-hydraulic two-phase transport of water, hydrogen,
14
C and
129
I
Authors: Enssle C. P. , Brommundt J. , Kaempfer Th. U. , Mayer G. , Wendling J.
Published: May 2014

The development of deep geological repositories for nuclear waste requires a sound system understanding, to which performance analyses by means of numerical flow and transport simulations form an important contribution. In this context, we present modelling studies of the thermo-hydraulic two-phase transport of water and hydrogen in the planned French repository in part 1. This paper, part 2, uses the same modelling concept extended to include radionuclide transport. The numerical TOUGH2-MP-model encompasses the host rock from cap to bed rock including the repository in full 3D. We performed a series of single-phase and two-phase transport simulations of volatile radionuclides (
14
C) and highly soluble ones (
129
I). The sensitivity analyses concerned different conditions and scenarios, for example, early canister failure, varied hydrogen generation regimes and transport parameters. The results allow determination of the dominant radionuclide transport paths and transfer processes. They demonstrate that hydrogen generation inside the repository has a large impact on the transfer of
14
C, favouring its advective transfer in the gas phase along the backfilled tunnels, whereas it has only minor influence on the transfer of
129
I which is mainly transported by diffusion in the formation water of the host rock. The study demonstrates the feasibility of full-scale radionuclide transport simulations.
Thyroid hormone inhibition in L6 myoblasts of IGF-I-mediated glucose uptake and proliferation: new roles for integrin αvβ3
Authors: Incerpi Sandra , Hsieh Meng-Ti , Lin Hung-Yun , Cheng Guei-Yun , De Vito Paolo , Fiore Anna Maria , Ahmed R. G. , Salvia Rosanna , Candelotti Elena , Leone Stefano , Luly Paolo , Pedersen Jens Z. , Davis Faith B. , Davis Paul J.
Published: May 2014

Thyroid hormones l-thyroxine (T4) and 3,3′,5-triiodo-l-thyronine (T3) have been shown to initiate short- and long-term effects via a plasma membrane receptor site located on integrin αvβ3. Also insulin-like growth factor type I (IGF-I) activity is known to be subject to regulation by this integrin. To investigate the possible cross-talk between T4 and IGF-I in rat L6 myoblasts, we have examined integrin αvβ3-mediated modulatory actions of T4 on glucose uptake, measured through carrier-mediated 2-deoxy-[3H]-d-glucose uptake, and on cell proliferation stimulated by IGF-I, assessed by cell counting, [3H]-thymidine incorporation, and fluorescence-activated cell sorting analysis. IGF-I stimulated glucose transport and cell proliferation via the cell surface IGF-I receptor (IGFIR) and, downstream of the receptor, by the phosphatidylinositol 3-kinase signal transduction pathway. Addition of 0.1 nM free T4 caused little or no cell proliferation but prevented both glucose uptake and proliferative actions of IGF-I. These actions of T4 were mediated by an Arg-Gly-Asp (RGD)-sensitive pathway, suggesting the existence of crosstalk between IGFIR and the T4 receptor located near the RGD recognition site on the integrin. An RGD-sequence-containing integrin inhibitor, a monoclonal antibody to αvβ3, and the T4 metabolite tetraiodothyroacetic acid all blocked the inhibition by T4 of IGF-I-stimulated glucose uptake and cell proliferation. Western blotting confirmed roles for activated phosphatidylinositol 3-kinase and extracellular regulated kinase 1/2 (ERK1/2) in the effects of IGF-I and also showed a role for ERK1/2 in the actions of T4 that modified the effects of IGF-I. We conclude that thyroid hormone inhibits IGF-I-stimulated glucose uptake and cell proliferation in L6 myoblasts.
Time window-dependent effect of perinatal maternal protein restriction on insulin sensitivity and energy substrate oxidation in adult male offspring
Authors: Agnoux Aurore Martin , Antignac Jean-Philippe , Simard Gilles , Poupeau Guillaume , Darmaun Dominique , Parnet Patricia , Alexandre-Gouabau Marie-Cécile
Published: May 2014

Epidemiological and experimental evidence suggests that a suboptimal environment during perinatal life programs offspring susceptibility to the development of metabolic syndrome and Type 2 diabetes. We hypothesized that the lasting impact of perinatal protein deprivation on mitochondrial fuel oxidation and insulin sensitivity would depend on the time window of exposure. To improve our understanding of underlying mechanisms, an integrative approach was used, combining the assessment of insulin sensitivity and untargeted mass spectrometry-based metabolomics in the offspring. A hyperinsulinemic-euglycemic clamp was performed in adult male rats born from dams fed a low-protein diet during gestation and/or lactation, and subsequently exposed to a Western diet (WD) for 10 wk. Metabolomics was combined with targeted acylcarnitine profiling and analysis of liver gene expression to identify markers of adaptation to WD that influence the phenotype outcome evaluated by body composition analysis. At adulthood, offspring of protein-restricted dams had impaired insulin secretion when fed a standard diet. Moreover, rats who demonstrated catch-up growth at weaning displayed higher gluconeogenesis and branched-chain amino acid catabolism, and lower fatty acid β-oxidation compared with control rats. Postweaning exposure of intrauterine growth restriction-born rats to a WD exacerbated incomplete fatty acid β-oxidation and excess fat deposition. Control offspring nursed by protein-restricted mothers showed peculiar low-fat accretion through adulthood and preserved insulin sensitivity even after WD-exposure. Altogether, our findings suggest a testable hypothesis about how maternal diet might influence metabolic outcomes (insulin sensitivity) in the next generation such as mitochondrial overload and/or substrate oxidation inflexibility dependent on the time window of perinatal dietary manipulation.
Low-dose B-type natriuretic peptide raises cardiac sympathetic nerve activity in sheep
Authors: Charles Christopher J. , Jardine David L. , Rademaker Miriam T. , Richards A. Mark
Published: May 2014

The reported effects of atrial natriuretic peptide (ANP) on sympathetic nerve activity (SNA) are variable, dependent on concomitant hemodynamic actions, and likely to be regionally differentiated. There are few reports of the effect of B-type natriuretic peptide (BNP) on SNA and none have measured cardiac SNA (CSNA) by direct microneurography. We measured the effects of low-dose ANP and BNP (2.4 pmol·kg−1·min−1 infused for 120 min) on CSNA and hemodynamics in conscious sheep ( n = 8). While there was a trend for mean arterial pressure and cardiac output to fall with both ANP and BNP, changes were not significant compared with vehicle control. However, BNP did significantly reduce systolic arterial (97 ± 4.2 vs. 107 ± 6.8 mmHg during control; P = 0.043) and pulse pressures (0.047) and increase heart rate (110 ± 6.7 vs. 96 ± 7.3 beats/min; P = 0.044). Trends for these hemodynamic parameters to change with ANP did not achieve statistical significance. ANP also had no significant effect on any CSNA parameters measured. In contrast, BNP induced a rise in both CSNA burst frequency (∼20 bursts/min higher than control, P = 0.011) and burst area (∼40% higher than control, P = 0.013). BNP-induced rises in burst incidence (bursts/100 beats), and burst area per 100 beats, however, were not significant. In conclusion, BNP infused at low doses that only had subtle effects on hemodynamics increased CSNA burst frequency and burst are per minute. This increase in CSNA may in large part be secondary to an increase in heart rate as CSNA burst incidence and burst area per 100 beats were not significantly increased. This study provides no evidence for inhibition of CSNA by natriuretic peptides.
Combination of bladder ultrasonography and novel cystometry method in mice reveals rapid decrease in bladder capacity and compliance in LPS-induced cystitis
Authors: Takezawa Kentaro , Kondo Makoto , Kiuchi Hiroshi , Soda Tetsuji , Takao Tetsuya , Miyagawa Yasushi , Tsujimura Akira , Nonomura Norio , Shimada Shoichi
Published: May 2014

Various animal models have been used in research into bladder dysfunction, and in vivo cystometry is a common method to analyze bladder function in animals. However, it is rather difficult to perform reliably in small animals. Transabdominal bladder ultrasonography combined with cystometry in urethane-anesthetized mice have revealed physical inhibition of bladder wall movement by a bladder catheter conventionally placed in the bladder apex. For reliable evaluation of mouse lower urinary tract function, we established a novel cystometry method in which a catheter was placed in the bladder anterior wall, in combination with bladder ultrasonography. This new method allowed the bladder to be well distended (i.e., larger maximum bladder capacity, lower pressure threshold, higher voided volume, and higher bladder compliance compared with conventional methods), which reflected more spontaneous voiding than conventional cystometry methods. We also demonstrated the usefulness of bladder ultrasonography for analysis of mouse bladder function, especially bladder dynamics, maximum bladder capacity, and post-voiding residual volume. We analyzed bladder functional changes in lipopolysaccharide (LPS)-induced cystitis by combining bladder ultrasonography and this new cystometry method. Bladder ultrasonography revealed a rapid decrease in bladder capacity, and cystometry showed a rapid decrease in voided volume due to intravesical LPS instillation. This new cystometry method also revealed a rapid decrease in bladder compliance caused by LPS instillation, which was not detectable by conventional methods. The combination of ultrasonography and the new cystometry method may become a powerful tool for analysis of mouse bladder function and could contribute to the development of new treatments for bladder dysfunction.
pH-responsive, gluconeogenic renal epithelial LLC-PK1-FBPase+cells: a versatile in vitro model to study renal proximal tubule metabolism and function
Authors: Curthoys Norman P. , Gstraunthaler Gerhard
Published: May 2014

Ammoniagenesis and gluconeogenesis are prominent metabolic features of the renal proximal convoluted tubule that contribute to maintenance of systemic acid-base homeostasis. Molecular analysis of the mechanisms that mediate the coordinate regulation of the two pathways required development of a cell line that recapitulates these features in vitro. By adapting porcine renal epithelial LLC-PK1cells to essentially glucose-free medium, a gluconeogenic subline, termed LLC-PK1-FBPase+cells, was isolated. LLC-PK1-FBPase+cells grow in the absence of hexoses and pentoses and exhibit enhanced oxidative metabolism and increased levels of phosphate-dependent glutaminase. The cells also express significant levels of the key gluconeogenic enzymes, fructose-1,6-bisphosphatase (FBPase) and phospho enolpyruvate carboxykinase (PEPCK). Thus the altered phenotype of LLC-PK1-FBPase+cells is pleiotropic. Most importantly, when transferred to medium that mimics a pronounced metabolic acidosis (9 mM HCO3−, pH 6.9), the LLC-PK1-FBPase+cells exhibit a gradual increase in NH4+ion production, accompanied by increases in glutaminase and cytosolic PEPCK mRNA levels and proteins. Therefore, the LLC-PK1-FBPase+cells retained in culture many of the metabolic pathways and pH-responsive adaptations characteristic of renal proximal tubules. The molecular mechanisms that mediate enhanced expression of the glutaminase and PEPCK in LLC-PK1-FBPase+cells have been extensively reviewed. The present review describes novel properties of this unique cell line and summarizes the molecular mechanisms that have been defined more recently using LLC-PK1-FBPase+cells to model the renal proximal tubule. It also identifies future studies that could be performed using these cells.
First D1-like receptor PET imaging of the rat and primate kidney: implications for human disease monitoring
Authors: Granda Michael L. , Schroeder Frederick A. , Borra Ronald H. J. , Schauer Nathan , Aisaborhale Ehimen , Guimaraes Alexander R. , Hooker Jacob M.
Published: May 2014

The intrarenal dopamine system is important for signaling and natriuresis, and significant dysfunction is associated with hypertension and kidney disease in ex vivo studies. Dopamine receptors also modulate and are modulated by the renin-angiotensin-aldosterone system. Here, we show the first in vivo measurement of D1-like receptors in the renal cortex of Sprague-Dawley rat and Papio anubis baboon using [11C]NNC 112, a positron emission tomography radioligand for D1-like receptors. In addition, we show a D1-like binding potential response to angiotensin II blockade in rats using losartan. Demonstration of self-saturable binding in the rat as well as specific and saturable binding in Papio anubis validate the use of [11C]NNC 112 in the first in vivo measurement of renal dopamine D1-like receptors. Furthermore, [11C]NNC 112 is a radioligand tool already validated for use in probing human central nervous system (CNS) D1-like receptors. Our work demonstrates specific and saturable non-CNS binding in higher animals and the ability to quantify physiological response to drug treatment and provides a clear path to extend use of [11C]NNC 112 to study renal dopamine in humans.
Novel mechanisms of Na+ retention in obesity: phosphorylation of NKCC2 and regulation of SPAK/OSR1 by AMPK
Authors: Davies Matthew , Fraser Scott A. , Galic Sandra , Choy Suet-Wan , Katerelos Marina , Gleich Kurt , Kemp Bruce E. , Mount Peter F. , Power David A.
Published: May 2014

Enhanced tubular reabsorption of salt is important in the pathogenesis of obesity-related hypertension, but the mechanisms remain poorly defined. To identify changes in the regulation of salt transporters in the kidney, C57BL/6 mice were fed a 40% fat diet [high-fat diet (HFD)] or a 12% fat diet (control diet) for 14 wk. Compared with control diet-fed mice, HFD-fed mice had significantly greater elevations in weight, blood pressure, and serum insulin and leptin levels. When we examined Na+ transporter expression, Na+-K+-2Cl− cotransporter (NKCC2) was unchanged in whole kidney and reduced in the cortex, Na+-Cl− cotransporter (NCC) and α-epithelial Na+ channel (ENaC) and γ-ENaC were unchanged, and β-ENaC was reduced. Phosphorylation of NCC was unaltered. Activating phosphorylation of NKCC2 at S126 was increased 2.5-fold. Activation of STE-20/SPS1-related proline-alanine-rich protein kinase (SPAK)/oxidative stress responsive 1 kinase (OSR1) was increased in kidneys from HFD-fed mice, and enhanced phosphorylation of NKCC2 at T96/T101 was evident in the cortex. Increased activity of NKCC2 in vivo was confirmed with diuretic experiments. HFD-fed mice had reduced activating phosphorylation of AMP-activated protein kinase (AMPK) in the renal cortex. In vitro, activation of AMPK led to a reduction in phospho-SPAK/phospho-OSR1 in AMPK+/+ murine embryonic fibroblasts (MEFs), but no effect was seen in AMPK−/− MEFs, indicating an AMPK-mediated effect. Activation of the with no lysine kinase/SPAK/OSR1 pathway with low-NaCl solution invoked a greater elevation in phospho-SPAK/phospho-OSR1 in AMPK−/− MEFs than in AMPK+/+ MEFs, consistent with a negative regulatory effect of AMPK on SPAK/OSR1 phosphorylation. In conclusion, this study identifies increased phosphorylation of NKCC2 on S126 as a hitherto-unrecognized mediator of enhanced Na+ reabsorption in obesity and identifies a new role for AMPK in regulating the activity of SPAK/OSR1.
Phosphate overload directly induces systemic inflammation and malnutrition as well as vascular calcification in uremia
Authors: Yamada Shunsuke , Tokumoto Masanori , Tatsumoto Narihito , Taniguchi Masatomo , Noguchi Hideko , Nakano Toshiaki , Masutani Kosuke , Ooboshi Hiroaki , Tsuruya Kazuhiko , Kitazono Takanari
Published: May 2014

Hyperphosphatemia contributes to increased cardiovascular mortality through vascular calcification (VC) in patients with chronic kidney disease (CKD). Malnutrition and inflammation are also closely linked to an increased risk of cardiovascular death in CKD. However, the effects of Pioverload on inflammation and malnutrition remain to be elucidated. The aim of the present study was to investigate the effects of dietary Piloading on the interactions among inflammation, malnutrition, and VC in CKD. We used control rats fed normal diets and adenine-induced CKD rats fed diets with different Piconcentrations ranging from 0.3% to 1.2% for 8 wk. CKD rats showed dietary Piconcentration-dependent increases in serum and tissue levels of TNF-α and urinary and tissue levels of oxidative stress markers and developed malnutrition (decrease in body weight, serum albumin, and urinary creatinine excretion), VC, and premature death without affecting kidney function. Treatment with 6% lanthanum carbonate blunted almost all changes induced by Pioverload. Regression analysis showed that serum Pilevels closely correlated with the extent of inflammation, malnutrition, and VC. Also, in cultured human vascular smooth muscle cells, high-Pimedium directly increased the expression of TNF-α in advance of the increase in osteochondrogenic markers. Our data suggest that dietary Pioverload induces systemic inflammation and malnutrition, accompanied by VC and premature death in CKD, and that inhibition of Piloading through dietary or pharmacological interventions or anti-inflammatory therapy may be a promising treatment for the prevention of malnutrition-inflammation-atherosclerosis syndrome.
Plane-specific Purkinje cell responses to vertical head rotations in the cat cerebellar nodulus and uvula
Authors: Kitama Toshihiro , Komagata Junya , Ozawa Kenichi , Suzuki Yutaka , Sato Yu
Published: May 2014

We recorded simple spike (SS) and complex spike (CS) firing of Purkinje cell in the cerebellar nodulus and uvula of awake, head-restrained cats during sinusoidal vertical rotation of the head in four stimulus planes (pitch, roll, and two vertical canal planes). Two SS response types (position- and velocity-types) with response phases close to those of head position and velocity, respectively, were recognized. Optimal response planes and directions for SS and CS of each cell were estimated from the response amplitudes in the four stimulus planes by fitting with a sinusoidal function. The principal findings are as follows: 1) two rostrocaudally oriented functional zones of Purkinje cells can be distinguished; 2) the medially located parasagittal band is active during rotation in the pitch plane; 3) the laterally located band is active during rotation in the roll plane. These two zones are the same as previously reported zones in the cerebellar flocculus active during head rotation in the canal planes in the point that both cerebellar sagittal zones are plane-specific functional zones, suggesting that the anatomical sagittal zones serve as functional plane-specific zones at least in the vestibulocerebellum.
Neural coding of passive lump detection in compliant artificial tissue
Authors: Gwilliam James C. , Yoshioka Takashi , Okamura Allison M. , Hsiao Steven S.
Published: May 2014

Here, we investigate the neural mechanisms of detecting lumps embedded in artificial compliant tissues. We performed a combined psychophysical study of humans performing a passive lump detection task with a neurophysiological study in nonhuman primates ( Macaca mulatta) where we recorded the responses of peripheral mechanoreceptive afferents to lumps embedded at various depths in intermediates (rubbers) of varying compliance. The psychophysical results reveal that human lump detection is greatly degraded by both lump depth and decreased compliance of the intermediate. The neurophysiology results reveal that only the slowly adapting type 1 (SA1) afferents provide a clear spatial representation of lumps at all depths and that the representation is affected by lump size, depth, and compliance of the intermediate. The rapidly adapting afferents are considerably less sensitive to the lump. We defined eight neural response measures that we hypothesized could explain the psychophysical behavior, including peak firing rate, spatial spread of neural activity, and additional parameters derived from these measures. We find that peak firing rate encodes the depth of the lump, and the neural spatial spread of the SA1 response encodes for lump size but not lump shape. We also find that the perception of lump size may be affected by the compliance of the intermediate. The results show that lump detection is based on a spatial population code of the SA1 afferents, which is distorted by the depth of the lump and compliance of the tissue.
Interference from mere thinking: mental rehearsal temporarily disrupts recall of motor memory
Authors: Yin Cong , Wei Kunlin
Published: May 2014

Interference between successively learned tasks is widely investigated to study motor memory. However, how simultaneously learned motor memories interact with each other has been rarely studied despite its prevalence in daily life. Assuming that motor memory shares common neural mechanisms with declarative memory system, we made unintuitive predictions that mental rehearsal, as opposed to further practice, of one motor memory will temporarily impair the recall of another simultaneously learned memory. Subjects simultaneously learned two sensorimotor tasks, i.e., visuomotor rotation and gain. They retrieved one memory by either practice or mental rehearsal and then had their memory evaluated. We found that mental rehearsal, instead of execution, impaired the recall of unretrieved memory. This impairment was content-independent, i.e., retrieving either gain or rotation impaired the other memory. Hence, conscious recollection of one motor memory interferes with the recall of another memory. This is analogous to retrieval-induced forgetting in declarative memory, suggesting a common neural process across memory systems. Our findings indicate that motor imagery is sufficient to induce interference between motor memories. Mental rehearsal, currently widely regarded as beneficial for motor performance, negatively affects memory recall when it is exercised for a subset of memorized items.
Adrenoreceptor modulation of oromotor pathways in the rat medulla
Authors: Nasse Jason S. , Travers Joseph B.
Published: May 2014

Regulation of feeding behavior involves the integration of multiple physiological and neurological pathways that control both nutrient-seeking and consummatory behaviors. The consummatory phase of ingestion includes stereotyped oromotor movements of the tongue and jaw that are controlled through brain stem pathways. These pathways encompass not only cranial nerve sensory and motor nuclei for processing feeding-related afferent signals and supplying the oromotor musculature but also reticular neurons for orchestrating ingestion and coordinating it with other behaviors that utilize the same musculature. Based on decerebrate studies, this circuit should be sensitive to satiety mechanisms mediated centrally by A2 noradrenergic neurons in the caudal nucleus of the solitary tract (cNST) that are potently activated during satiety. Because the first observable phase of satiety is inhibition of oromotor movements, we hypothesized that norepinephrine (NE) would act to inhibit prehypoglossal neurons in the medullary reticular formation. Using patch-clamp electrophysiology of retrogradely labeled prehypoglossal neurons and calcium imaging to test this hypothesis, we demonstrate that norepinephrine can influence both pre- and postsynaptic properties of reticular neurons through both α1- and α2-adrenoreceptors. The α1-adrenoreceptor agonist phenylephrine (PE) activated an inward current in the presence of TTX and increased the frequency of both inhibitory and excitatory miniature postsynaptic currents. The α2-adrenoreceptor agonist dexmedetomidine (DMT) inhibited cNST-evoked excitatory currents as well as spontaneous and miniature excitatory currents through presynaptic mechanisms. The diversity of adrenoreceptor modulation of these prehypoglossal neurons may reflect their role in a multifunctional circuit coordinating both ingestive and respiratory lingual function.
Characterization of motor units in behaving adult mice shows a wide primary range
Authors: Ritter Laura K. , Tresch Matthew C. , Heckman C. J. , Manuel Marin , Tysseling Vicki M.
Published: May 2014

The mouse is essential for genetic studies of motor function in both normal and pathological states. Thus it is important to consider whether the structure of motor output from the mouse is in fact analogous to that recorded in other animals. There is a striking difference in the basic electrical properties of mouse motoneurons compared with those in rats, cats, and humans. The firing evoked by injected currents produces a unique frequency-current (F-I) function that emphasizes recruitment of motor units at their maximum force. These F-I functions, however, were measured in anesthetized preparations that lacked two key components of normal synaptic input: high levels of synaptic noise and neuromodulatory inputs. Recent studies suggest that the alterations in the F-I function due to these two components are essential for recreating firing behavior of motor units in human subjects. In this study we provide the first data on firing patterns of motor units in the awake mouse, focusing on steady output in quiet stance. The resulting firing patterns did not match the predictions from the mouse F-I behaviors but instead revealed rate modulation across a remarkably wide range (10–60 Hz). The low end of the firing range may be due to changes in the F-I relation induced by synaptic noise and neuromodulatory inputs. The high end of the range may indicate that, unlike other species, quiet standing in the mouse involves recruitment of relatively fast-twitch motor units.
A Multiobjective Iterated Greedy Algorithm for Truck Scheduling in Cross-Dock Problems
Authors: Naderi B. , Rahmani Shadi , Rahmani Shabnam
Published: May 2014

The cross-docking system is a new distribution strategy which can reduce inventories, lead times, and improve responding time to customers. This paper considers biobjective problem of truck scheduling in cross-docking systems with temporary storage. The objectives are minimizing both makespan and total tardiness. For this problem, it proposes a multiobjective iterated greedy algorithm employing advance features such as modified crowding selection, restart phase, and local search. To evaluate the proposed algorithm for performance, it is compared with two available algorithms, subpopulation particle swarm optimization-II and strength Pareto evolutionary algorithm-II. The comparison shows that the proposed multiobjective iterated greedy algorithm shows high performance and outperforms the other two algorithms.