bluuebunny/crossref_metadata_2025 huggingface dataset:
Modulation of spinal cord synaptic activity by tumor necrosis factor α in a model of peripheral neuropathy
Authors: Spicarova Diana , Nerandzic Vladimir , Palecek Jiri
Published: December 2011

Background

The cytokine tumor necrosis factor α (TNFα) is an established pain modulator in both the peripheral and central nervous systems. Modulation of nociceptive synaptic transmission in the spinal cord dorsal horn (DH) is thought to be involved in the development and maintenance of several pathological pain states. Increased levels of TNFα and its receptors (TNFR) in dorsal root ganglion (DRG) cells and in the spinal cord DH have been shown to play an essential role in neuropathic pain processing. In the present experiments the effect of TNFα incubation on modulation of primary afferent synaptic activity was investigated in a model of peripheral neuropathy.

Methods

Spontaneous and miniature excitatory postsynaptic currents (sEPSC and mEPSCs) were recorded in superficial DH neurons in acute spinal cord slices prepared from animals 5 days after sciatic nerve transection and in controls.

Results

In slices after axotomy the sEPSC frequency was 2.8 ± 0.8 Hz, while neurons recorded from slices after TNFα incubation had significantly higher sEPSC frequency (7.9 ± 2.2 Hz). The effect of TNFα treatment was smaller in the slices from the control animals, where sEPSC frequency was 1.2 ± 0.2 Hz in slices without and 2.0 ± 0.5 Hz with TNFα incubation. Tetrodotoxin (TTX) application in slices from axotomized animals and after TNFα incubation decreased the mEPSC frequency to only 37.4 ± 6.9% of the sEPSC frequency. This decrease was significantly higher than in the slices without the TNFα treatment (64.4 ± 6.4%). TTX application in the control slices reduced the sEPSC frequency to about 80% in both TNFα untreated and treated slices. Application of low concentration TRPV1 receptors endogenous agonist N-oleoyldopamine (OLDA, 0.2 μM) in slices after axotomy induced a significant increase in mEPSC frequency (175.9 ± 17.3%), similar to the group with TNFα pretreatment (158.1 ± 19.5%).

Conclusions

Our results indicate that TNFα may enhance spontaneous transmitter release from primary afferent fibres in the spinal cord DH by modulation of TTX-sensitive sodium channels following sciatic nerve transection. This nerve injury also leads to enhanced sensitivity of presynaptic TRPV1 receptors to endogenous agonist. Modulation of presynaptic receptor activity on primary sensory terminals by TNFα may play an important role in neuropathic pain development.
Diversity and phylogeography of begomovirus-associated beta satellites of okra in India
Authors: Venkataravanappa V , Lakshminarayana Reddy CN , Swaranalatha P , Jalali Salil , Briddon Rob W , Reddy M Krishna
Published: December 2011

Background

Okra (Abelmoschus esculentus; family Malvaceae) is grown in temperate as well as subtropical regions of the world, both for human consumption as a vegetable and for industrial uses. Okra yields are affected by the diseases caused by phyopathogenic viruses. India is the largest producer of okra and in this region a major biotic constraint to production are viruses of the genus Begomovirus. Begomoviruses affecting okra across the Old World are associated with specific, symptom modulating satellites (beta satellites). We describe a comprehensive analysis of the diversity of beta satellites associated with okra in India.

Results

The full-length sequences of 36 beta satellites, isolated from okra exhibiting typical begomovirus symptoms (leaf curl and yellow vein), were determined. The sequences segregated in to four groups. Two groups correspond to the beta satellites Okra leaf curl beta satellite (OLCuB) and Bhendi yellow vein beta satellite (BYVB) that have previously been identified in okra from the sub-continent. One sequence was distinct from all other, previously isolated beta satellites and represents a new species for which we propose the name Bhendi yellow vein India beta satellite (BYVIB). This new beta satellite was nevertheless closely related to BYVB and OLCuB. Most surprising was the identification of Croton yellow vein mosaic beta satellite (CroYVMB) in okra; a beta satellite not previously identified in a malvaceous plant species. The okra beta satellites were shown to have distinct geographic host ranges with BYVB occurring across India whereas OLCuB was only identified in northwestern India. Okra infections with CroYVMB were only identified across the northern and eastern central regions of India. A more detailed analysis of the sequences showed that OLCuB, BYVB and BYVIB share highest identity with respect βC1 gene. βC1 is the only gene encoded by beta satellites, the product of which is the major pathogenicity determinant of begomovirus-beta satellite complexes and is involved in overcoming host defenses based on RNAi.

Conclusion

The diversity of beta satellites in okra across the sub-continent is higher than previously realized and is higher than for any other malvaceous plant species so far analyzed. The beta satellites identified in okra show geographic segregation, which has implications for the development and introduction of resistant okra varieties. However, the finding that the βC1 gene of the major okra beta satellites (OLCuB, BYVB and BYVIB) share high sequence identity and provides a possible avenue to achieve a broad spectrum resistance.
CRISPR loci reveal networks of gene exchange in archaea
Authors: Brodt Avital , Lurie-Weinberger Mor N , Gophna Uri
Published: December 2011

Background

CRISPR (Clustered, Regularly, Interspaced, Short, Palindromic Repeats) loci provide prokaryotes with an adaptive immunity against viruses and other mobile genetic elements. CRISPR arrays can be transcribed and processed into small crRNA molecules, which are then used by the cell to target the foreign nucleic acid. Since spacers are accumulated by active CRISPR/Cas systems, the sequences of these spacers provide a record of the past "infection history" of the organism.

Results

Here we analyzed all currently known spacers present in archaeal genomes and identified their source by DNA similarity. While nearly 50% of archaeal spacers matched mobile genetic elements, such as plasmids or viruses, several others matched chromosomal genes of other organisms, primarily other archaea. Thus, networks of gene exchange between archaeal species were revealed by the spacer analysis, including many cases of inter-genus and inter-species gene transfer events. Spacers that recognize viral sequences tend to be located further away from the leader sequence, implying that there exists a selective pressure for their retention.

Conclusions

CRISPR spacers provide direct evidence for extensive gene exchange in archaea, especially within genera, and support the current dogma where the primary role of the CRISPR/Cas system is anti-viral and anti-plasmid defense.

Open peer review

This article was reviewed by: Profs. W. Ford Doolittle, John van der Oost, Christa Schleper (nominated by board member Prof. J Peter Gogarten)
A rat model of early stage osteonecrosis induced by glucocorticoids
Authors: Kerachian Mohammad Amin , Harvey Edward J , Cournoyer Denis , Chow Terry Y , Nahal Ayoub , Séguin Chantal
Published: December 2011

Background

Glucocorticoid (GC)-induced osteonecrosis (ON) is an important complication of medical therapy. The exact pathomechanisms of ON has not been clearly elucidated. There is a need for a reproducible animal model that better approximates the clinical scenario.

Methods

To determine the genetic susceptibility of rats to develop GC-induced femoral head ON, we evaluated 5 different inbred strains of rats (Spontaneous Hypertensive Rat, Wistar Kyoto, Wistar Furth, SASCO Fisher and Lewis). Prednisone pellets (dosage of 1.82-2.56 mg/kg/day) were implanted subcutaneously for 90. After 90 days, the femurs were resected and examined histologically and radiographically. Pathological and histological examination was performed. Hematoxylin and eosin (H & E) staining was used to delineate the femoral head osteonecrosis lesions as well as abnormalities of articular cartilage and growth plate.

Results

The greatest differences in H & E staining were seen in the Wistar Kyoto and Wistar Furth groups. In these groups 4 out of 5 and 3 out of 5, respectively, steroid-induced rats revealed growth plate disruption with acellular areas. The TUNEL apoptosis staining assay for apoptosis revealed that 4 out of 5 of Wistar Kyoto rats, 5 out of 5 of Wistar Furth, 2 out of 4 of surviving Lewis and 2 out of 2 of the surviving spontaneous hypertensive rats had apoptotic osteocytes in trabeculae, whereas none of the Fisher rats showed apoptotic osteocytes.

Conclusions

We postulate that Wistar Kyoto, Wistar Furth and spontaneous hypertensive rats may be strains of rats more susceptible to develop ON of the femoral head while Fisher rats were the most resistant.
A novel mean-centering method for normalizing microRNA expression from high-throughput RT-qPCR data
Authors: Wylie Dennis , Shelton Jeffrey , Choudhary Ashish , Adai Alex T
Published: December 2011

Background

Normalization is critical for accurate gene expression analysis. A significant challenge in the quantitation of gene expression from biofluids samples is the inability to quantify RNA concentration prior to analysis, underscoring the need for robust normalization tools for this sample type. In this investigation, we evaluated various methods of normalization to determine the optimal approach for quantifying microRNA (miRNA) expression from biofluids and tissue samples when using the TaqMan® Megaplex™ high-throughput RT-qPCR platform with low RNA inputs.

Findings

We compared seven normalization methods in the analysis of variation of miRNA expression from biofluid and tissue samples. We developed a novel variant of the common mean-centering normalization strategy, herein referred to as mean-centering restricted (MCR) normalization, which is adapted to the TaqMan Megaplex RT-qPCR platform, but is likely applicable to other high-throughput RT-qPCR-based platforms. Our results indicate that MCR normalization performs comparable to or better than both standard mean-centering and other normalization methods. We also propose an extension of this method to be used when migrating biomarker signatures from Megaplex to singleplex RT-qPCR platforms, based on the identification of a small number of normalizer miRNAs that closely track the mean of expressed miRNAs.

Conclusions

We developed the MCR method for normalizing miRNA expression from biofluids samples when using the TaqMan Megaplex RT-qPCR platform. Our results suggest that normalization based on the mean of all fully observed (fully detected) miRNAs minimizes technical variance in normalized expression values, and that a small number of normalizer miRNAs can be selected when migrating from Megaplex to singleplex assays. In our study, we find that normalization methods that focus on a restricted set of miRNAs tend to perform better than methods that focus on all miRNAs, including those with non-determined (missing) values. This methodology will likely be most relevant for studies in which a significant number of miRNAs are not detected.
Neuromelanin, neurotransmitter status and brainstem location determine the differential vulnerability of catecholaminergic neurons to mitochondrial DNA deletions
Authors: Elstner Matthias , Müller Sarina K , Leidolt Lars , Laub Christoph , Krieg Lena , Schlaudraff Falk , Liss Birgit , Morris Chris , Turnbull Douglass M , Masliah Eliezer , Prokisch Holger , Klopstock Thomas , Bender Andreas
Published: December 2011

Background

Deletions of the mitochondrial DNA (mtDNA) accumulate to high levels in dopaminergic neurons of the substantia nigra pars compacta (SNc) in normal aging and in patients with Parkinson's disease (PD). Human nigral neurons characteristically contain the pigment neuromelanin (NM), which is believed to alter the cellular redox-status. The impact of neuronal pigmentation, neurotransmitter status and brainstem location on the susceptibility to mtDNA damage remains unclear. We quantified mtDNA deletions (ΔmtDNA) in single pigmented and non-pigmented catecholaminergic, as well as non-catecholaminergic neurons of the human SNc, the ventral tegmental area (VTA) and the locus coeruleus (LC), using laser capture microdissection and single-cell real-time PCR.

Results

In healthy aged individuals, ΔmtDNA levels were highest in pigmented catecholaminergic neurons (25.2 ± 14.9%), followed by non-pigmented catecholamergic (18.0 ± 11.2%) and non-catecholaminergic neurons (12.3 ± 12.3%; p < 0.001). Within the catecholaminergic population, ΔmtDNA levels were highest in dopaminergic neurons of the SNc (33.9 ± 21.6%) followed by dopaminergic neurons of the VTA (21.9 ± 12.3%) and noradrenergic neurons of the LC (11.1 ± 11.4%; p < 0.001). In PD patients, there was a trend to an elevated mutation load in surviving non-pigmented nigral neurons (27.13 ± 16.73) compared to age-matched controls (19.15 ± 11.06; p = 0.052), but levels where similar in pigmented nigral neurons of PD patients (41.62 ± 19.61) and controls (41.80 ± 22.62).

Conclusions

Catecholaminergic brainstem neurons are differentially susceptible to mtDNA damage. Pigmented dopaminergic neurons of the SNc show the highest ΔmtDNA levels, possibly explaining the exceptional vulnerability of the nigro-striatal system in PD and aging. Although loss of pigmented noradrenergic LC neurons also is an early feature of PD pathology, mtDNA levels are not elevated in this nucleus in healthy controls. Thus, ΔmtDNA are neither an inevitable consequence of catecholamine metabolism nor a universal explanation for the regional vulnerability seen in PD.
Adaptation of Rhizobium leguminosarumto pea, alfalfa and sugar beet rhizospheres investigated by comparative transcriptomics
Authors: Ramachandran Vinoy K , East Alison K , Karunakaran Ramakrishnan , Downie J Allan , Poole Philip S
Published: December 2011

Background

The rhizosphere is the microbe-rich zone around plant roots and is a key determinant of the biosphere's productivity. Comparative transcriptomics was used to investigate general and plant-specific adaptations during rhizosphere colonization. Rhizobium leguminosarum biovar viciae was grown in the rhizospheres of pea (its legume nodulation host), alfalfa (a non-host legume) and sugar beet (non-legume). Gene expression data were compared to metabolic and transportome maps to understand adaptation to the rhizosphere.

Results

Carbon metabolism was dominated by organic acids, with a strong bias towards aromatic amino acids, C1 and C2 compounds. This was confirmed by induction of the glyoxylate cycle required for C2 metabolism and gluconeogenesis in all rhizospheres. Gluconeogenesis is repressed in R. leguminosarum by sugars, suggesting that although numerous sugar and putative complex carbohydrate transport systems are induced in the rhizosphere, they are less important carbon sources than organic acids. A common core of rhizosphere-induced genes was identified, of which 66% are of unknown function. Many genes were induced in the rhizosphere of the legumes, but not sugar beet, and several were plant specific. The plasmid pRL8 can be considered pea rhizosphere specific, enabling adaptation of R. leguminosarum to its host. Mutation of many of the up-regulated genes reduced competitiveness for pea rhizosphere colonization, while two genes specifically up-regulated in the pea rhizosphere reduced colonization of the pea but not alfalfa rhizosphere.

Conclusions

Comparative transcriptome analysis has enabled differentiation between factors conserved across plants for rhizosphere colonization as well as identification of exquisite specific adaptation to host plants.
Curated collection of yeast transcription factor DNA binding specificity data reveals novel structural and gene regulatory insights
Authors: Gordân Raluca , Murphy Kevin F , McCord Rachel P , Zhu Cong , Vedenko Anastasia , Bulyk Martha L
Published: December 2011

Background

Transcription factors (TFs) play a central role in regulating gene expression by interacting with cis-regulatory DNA elements associated with their target genes. Recent surveys have examined the DNA binding specificities of most Saccharomyces cerevisiae TFs, but a comprehensive evaluation of their data has been lacking.

Results

We analyzed in vitro and in vivo TF-DNA binding data reported in previous large-scale studies to generate a comprehensive, curated resource of DNA binding specificity data for all characterized S. cerevisiae TFs. Our collection comprises DNA binding site motifs and comprehensive in vitro DNA binding specificity data for all possible 8-bp sequences. Investigation of the DNA binding specificities within the basic leucine zipper (bZIP) and VHT1 regulator (VHR) TF families revealed unexpected plasticity in TF-DNA recognition: intriguingly, the VHR TFs, newly characterized by protein binding microarrays in this study, recognize bZIP-like DNA motifs, while the bZIP TF Hac1 recognizes a motif highly similar to the canonical E-box motif of basic helix-loop-helix (bHLH) TFs. We identified several TFs with distinct primary and secondary motifs, which might be associated with different regulatory functions. Finally, integrated analysis of in vivo TF binding data with protein binding microarray data lends further support for indirect DNA binding in vivo by sequence-specific TFs.

Conclusions

The comprehensive data in this curated collection allow for more accurate analyses of regulatory TF-DNA interactions, in-depth structural studies of TF-DNA specificity determinants, and future experimental investigations of the TFs' predicted target genes and regulatory roles.
Disruption of the Chemokine-Like Receptor-1 (CMKLR1) Gene Is Associated with Reduced Adiposity and Glucose Intolerance
Authors: Ernst Matthew C. , Haidl Ian D. , Zúñiga Luis A. , Dranse Helen J. , Rourke Jillian L. , Zabel Brian A. , Butcher Eugene C. , Sinal Christopher J.
Published: December 2011

Adipose tissue secretes a variety of bioactive signaling molecules, termed adipokines, which regulate numerous biological functions including appetite, energy balance, glucose homeostasis, and inflammation. Chemerin is a novel adipokine that regulates adipocyte differentiation and metabolism by binding to and activating the G protein-coupled receptor, chemokine like receptor-1 (CMKLR1). In the present study, we investigated the impact of CMKLR1 deficiency on adipose development, glucose homeostasis, and inflammation in vivo. Herein we report that regardless of diet (low or high fat), CMKLR1−/− mice had lower food consumption, total body mass, and percent body fat compared with wild-type controls. CMKLR1−/− mice also exhibited decreased hepatic and white adipose tissue TNFα and IL-6 mRNA levels coincident with decreased hepatic dendritic cell infiltration, decreased adipose CD3+ T cells, and increased adipose natural killer cells. CMKLR1−/− mice were glucose intolerant compared with wild-type mice, and this was associated with decreased glucose stimulated insulin secretion as well as decreased skeletal muscle and white adipose tissue glucose uptake. Collectively these data provide compelling evidence that CMKLR1 influences adipose tissue development, inflammation, and glucose homeostasis and may contribute to the metabolic derangement characteristic of obesity and obesity-related diseases.
Novel Biological Action of the Dipeptidylpeptidase-IV Inhibitor, Sitagliptin, as a Glucagon-Like Peptide-1 Secretagogue
Authors: Sangle Ganesh V. , Lauffer Lina M. , Grieco Anthony , Trivedi Shivangi , Iakoubov Roman , Brubaker Patricia L.
Published: December 2011

Glucagon-like peptide-1 (GLP-1) is an incretin hormone secreted into the circulation by the intestinal L cell. The dipeptidylpeptidase-IV (DPP-IV) inhibitor, sitagliptin, prevents GLP-1 degradation and is used in the clinic to treat patients with type 2 diabetes mellitus, leading to improved glycated hemoglobin levels. When the effect of sitagliptin on GLP-1 levels was examined in neonatal streptozotocin rats, a model of type 2 diabetes mellitus, a 4.9 ± 0.9-fold increase in basal and 3.6 ± 0.4-fold increase in oral glucose-stimulated plasma levels of active GLP-1 was observed (P < 0.001), in association with a 1.5 ± 0.1-fold increase in the total number of intestinal L cells (P < 0.01). The direct effects of sitagliptin on GLP-1 secretion and L cell signaling were therefore examined in murine GLUTag (mGLUTag) and human hNCI-H716 intestinal L cells in vitro. Sitagliptin (0.1–2 μm) increased total GLP-1 secretion by mGLUTag and hNCI-H716 cells (P < 0.01–0.001). However, MK0626 (1–50 μm), a structurally unrelated inhibitor of DPP-IV, did not affect GLP-1 secretion in either model. Treatment of mGLUTag cells with the GLP-1 receptor agonist, exendin-4, did not modulate GLP-1 release, indicating the absence of feedback effects of GLP-1 on the L cell. Sitagliptin increased cAMP levels (P < 0.01) and ERK1/2 phosphorylation (P < 0.05) in both mGLUTag and hNCI-H716 cells but did not alter either intracellular calcium or phospho-Akt levels. Pretreatment of mGLUTag cells with protein kinase A (H89 and protein kinase inhibitor) or MAPK kinase-ERK1/2 (PD98059 and U0126) inhibitors prevented sitagliptin-induced GLP-1 secretion (P < 0.05–0.01). These studies demonstrate, for the first time, that sitagliptin exerts direct, DPP-IV-independent effects on intestinal L cells, activating cAMP and ERK1/2 signaling and stimulating total GLP-1 secretion.
Corncob Bedding Alters the Effects of Estrogens on Aggressive Behavior and Reduces Estrogen Receptor-α Expression in the Brain
Authors: Landeros Rosalina Villalon , Morisseau Christophe , Yoo Hyun Ju , Fu Samuel H. , Hammock Bruce D. , Trainor Brian C.
Published: December 2011

There is growing appreciation that estrogen signaling pathways can be modulated by naturally occurring environmental compounds such as phytoestrogens and the more recently discovered xenoestrogens. Many researchers studying the effects of estrogens on brain function or behavior in animal models choose to use phytoestrogen-free food for this reason. Corncob bedding is commonly used in animal facilities across the United States and has been shown to inhibit estrogen-dependent reproductive behavior in rats. The mechanism for this effect was unclear, because the components of corncob bedding mediating this effect did not bind estrogen receptors. Here, we show in the California mouse (Peromyscus californicus) that estrogens decrease aggression when cardboard-based bedding is used but that this effect is absent when corncob bedding is used. California mice housed on corncob bedding also had fewer estrogen receptor-α-positive cells in the bed nucleus of the stria terminalis and ventromedial hypothalamus compared with mice housed on cardboard-based bedding. In addition, corncob bedding suppressed the expression of phosphorylated ERK in these brain regions as well as in the medial amygdala and medial preoptic area. Previous reports of the effects of corncob bedding on reproductive behavior are not widely appreciated. Our observations on the effects of corncob bedding on behavior and brain function should draw attention to the importance that cage bedding can exert on neuroendocrine research.
Integrating perspectives on vocal performance and consistency
Authors: Sakata Jon T. , Vehrencamp Sandra L.
Published: December 2011

Recent experiments in divergent fields of birdsong have revealed that vocal performance is important for reproductive success and under active control by distinct neural circuits. Vocal consistency, the degree to which the spectral properties (e.g. dominant or fundamental frequency) of song elements are produced consistently from rendition to rendition, has been highlighted as a biologically important aspect of vocal performance. Here, we synthesize functional, developmental and mechanistic (neurophysiological) perspectives to generate an integrated understanding of this facet of vocal performance. Behavioral studies in the field and laboratory have found that vocal consistency is affected by social context, season and development, and, moreover, positively correlated with reproductive success. Mechanistic investigations have revealed a contribution of forebrain and basal ganglia circuits and sex steroid hormones to the control of vocal consistency. Across behavioral, developmental and mechanistic studies, a convergent theme regarding the importance of vocal practice in juvenile and adult songbirds emerges, providing a basis for linking these levels of analysis. By understanding vocal consistency at these levels, we gain an appreciation for the various dimensions of song control and plasticity and argue that genes regulating the function of basal ganglia circuits and sex steroid hormones could be sculpted by sexual selection.
Interactions between cortisol and Rhesus glycoprotein expression in ureogenic toadfish, Opsanus beta
Authors: Rodela Tamara M. , McDonald M. Danielle , Walsh Patrick J. , Gilmour Kathleen M.
Published: December 2011

In their native environment, gulf toadfish excrete equal quantities of ammonia and urea. However, upon exposure to stressful conditions in the laboratory (i.e. crowding, confinement or air exposure), toadfish decrease branchial ammonia excretion and become ureotelic. The objective of this study was to determine the influences of cortisol and ammonia on ammonia excretion relative to expression of Rhesus (Rh) glycoproteins and the ammonia-fixing enzyme, glutamine synthetase (GS). In vivo infusions and/or injections were used to manipulate corticosteroid activity and plasma ammonia concentrations in ureotelic toadfish. Metyrapone treatment to lower circulating cortisol levels resulted in a 3.5-fold elevation of ammonia excretion rates, enhanced mRNA expression of two of the toadfish Rh isoforms (Rhcg1 and Rhcg2), and decreased branchial and hepatic GS activity. Correspondingly, cortisol infusion decreased ammonia excretion 2.5-fold, a change that was accompanied by reduced branchial expression of all toadfish Rh isoforms (Rhag, Rhbg, Rhcg1 and Rhcg2) and a twofold increase in hepatic GS activity. In contrast, maintenance of high circulating ammonia levels by ammonia infusion enhanced ammonia excretion and Rh expression (Rhag, Rhbg and Rhcg2). Toadfish treated with cortisol showed an attenuated response to ammonia infusion with no change in Rh mRNA expression or GS activity. In summary, the evidence suggests that ammonia excretion in toadfish is modulated by cortisol-induced changes in both Rh glycoprotein expression and GS activity.
Evidence for damage-dependent hygienic behaviour towards Varroa destructor-parasitised brood in the western honey bee, Apis mellifera
Authors: Schöning Caspar , Gisder Sebastian , Geiselhardt Sven , Kretschmann Ivonne , Bienefeld Kaspar , Hilker Monika , Genersch Elke
Published: December 2011

The ectoparasitic mite Varroa destructor and honey bee pathogenic viruses have been implicated in the recent demise of honey bee colonies. Several studies have shown that the combination of V. destructor and deformed wing virus (DWV) poses an especially serious threat to honey bee health. Mites transmitting virulent forms of DWV may cause fatal DWV infections in the developing bee, while pupae parasitised by mites not inducing or activating overt DWV infections may develop normally. Adult bees respond to brood diseases by removing affected brood. This hygienic behaviour is an essential part of the bees' immune response repertoire and is also shown towards mite-parasitised brood. However, it is still unclear whether the bees react towards the mite in the brood cell or rather towards the damage done to the brood. We hypothesised that the extent of mite-associated damage rather than the mere presence of parasitising mites triggers hygienic behaviour. Hygienic behaviour assays performed with mites differing in their potential to transmit overt DWV infections revealed that brood parasitised by ‘virulent’ mites (i.e. mites with a high potential to induce fatal DWV infections in parasitised pupae) were removed significantly more often than brood parasitised by ‘less virulent’ mites (i.e. mites with a very low potential to induce overt DWV infections) or non-parasitised brood. Chemical analyses of brood odour profiles suggested that the bees recognise severely affected brood by olfactory cues. Our results suggest that bees show selective, damage-dependent hygienic behaviour, which may be an economic way for colonies to cope with mite infestation.
Magnesium chenoursodeoxycholic acid ameliorates carbon tetrachloride-induced liver fibrosis in rats
Authors: Kang Jung-Woo , Yoon Seong-Jin , Sung Yong-Kyung , Lee Sun-Mee
Published: December 2011

This study was designed to investigate the hepatoprotective effects of magnesium chenoursodeoxycholic acid (Mg-CUD), a magnesium trihydrate salt of ursodeoxycholic acid and chenodeoxycholic acid, in carbon tetrachloride (CCl4)-induced liver fibrosis in rats. Rats were treated with CCl4 dissolved in olive oil (0.5 mL/kg, twice a week) intraperitoneally for eight weeks. Mg-CUD was administered orally at 15.625, 31.25, 62.5 and 125 mg/kg once a day. Chronic CCl4 administration induced increases in serum transforming growth factor- β1, hepatic hydroxyproline content and serum alanine aminotransferase activity. Mg-CUD attenuated these increases. The levels of α-smooth muscle actin protein and mRNA expression were increased by chronic CCl4 exposure and Mg-CUD attenuated these increases. Mg-CUD suppressed increases in matrix metalloproteinase-2 and tissue inhibitor of metalloproteinase-1 mRNA expression and elevation of oxidative stresses by attenuating lipid peroxidation and enhancing reduced glutathione/oxidized glutathione ratio. The overexpression of toll-like receptor 4 and increased nuclear translocation of nuclear factor- κB and phosphorylated c-Jun, a component of activator protein 1, were suppressed by Mg-CUD. Furthermore, CCl4 increased the levels of tumor necrosis factor (TNF)- α, interleukin (IL)-6, IL-10 and cyclooxygenase (COX)-2. Mg-CUD attenuated the levels of TNF- α, IL-6 and COX-2, while it augmented the level of IL-10. Our results suggest that Mg-CUD may prevent liver fibrosis by modulating collagen accumulation and inflammatory signaling pathways.
Crystal structure of release factor RF3 trapped in the GTP state on a rotated conformation of the ribosome
Authors: Zhou Jie , Lancaster Laura , Trakhanov Sergei , Noller Harry F.
Published: December 2011

The class II release factor RF3 is a GTPase related to elongation factor EF-G, which catalyzes release of class I release factors RF1 and RF2 from the ribosome after termination of protein synthesis. The 3.3 Å crystal structure of the RF3·GDPNP·ribosome complex provides a high-resolution description of interactions and structural rearrangements that occur when binding of this translational GTPase induces large-scale rotational movements in the ribosome. RF3 induces a 7° rotation of the body and 14° rotation of the head of the 30S ribosomal subunit, and itself undergoes inter- and intradomain conformational rearrangements. We suggest that ordering of critical elements of switch loop I and the P loop, which help to form the GTPase catalytic site, are caused by interactions between the G domain of RF3 and the sarcin–ricin loop of 23S rRNA. The rotational movements in the ribosome induced by RF3, and its distinctly different binding orientation to the sarcin–ricin loop of 23S rRNA, raise interesting implications for the mechanism of action of EF-G in translocation.
Quantifying Transaction Costs in Online/Off-line Grocery Channel Choice
Authors: Chintagunta Pradeep K. , Chu Junhong , Cebollada Javier
Published: December 2011

Households incur transaction costs when choosing among off-line stores for grocery purchases. They may incur additional transaction costs when buying groceries online versus off-line. We integrate the various transaction costs into a channel choice framework and empirically quantify the relative transaction costs when households choose between the online and off-line channels of the same grocery chain. The key challenges in quantifying these costs are (i) the complexity of channel choice decision and (ii) that several of the costs depend on the items a household expects to buy in the store, and unobserved factors that influence channel choice also likely influence the items purchased. We use the unique features of our empirical context to address the first issue and the plausibly exogenous approach in a hierarchical Bayesian framework to account for the endogeneity of the channel choice drivers. We find that transaction costs for grocery shopping can be sizable and play an important role in the choice between online and off-line channels. We provide monetary metrics for several types of transaction costs, such as travel time and transportation costs, in-store shopping time, item-picking costs, basket-carrying costs, quality inspection costs, and inconvenience costs. We find considerable household heterogeneity in these costs and characterize their distributions. We discuss the implications of our findings for the retailer's channel strategy.
Customer Bill of Rights Under No-Fault Service Failure: Confinement and Compensation
Authors: Chen Rachel R. , Gerstner Eitan , Yang Yinghui (Catherine)
Published: December 2011

Service providers and their customers are sometimes victims of failures caused by exogenous factors such as unexpected bad weather, power outages, or labor strikes. When such no-fault failures occur in confined zones, service providers may confine customers against their will if making arrangements for them to leave is very costly. Such confinements, however, can result in severe pain and suffering, and customer complaints put regulators under pressure to pass a customer bill of rights that allows captive customers to abort failed services. This paper shows that service providers are better off preempting such laws by voluntarily allowing customers to escape the service under failure. Moreover, service providers can profit by targeting compensation to customers based on whether they use or leave the service under failure.
The challenges of evaluating large-scale, multi-partner programmes: the case of NIHR CLAHRCs
Authors: Martin Graham P. , Ward Vicky , Hendy Jane , Rowley Emma , Nancarrow Susan , Heaton Janet , Britten Nicky , Fielden Sandra , Ariss Steven
Published: December 2011

The limited extent to which research evidence is utilised in healthcare and other public services is widely acknowledged. The United Kingdom government has attempted to address this gap by funding nine Collaborations for Leadership in Applied Health Research and Care (CLAHRCs). CLAHRCs aim to carry out health research, implement research findings in local healthcare organisations and build capacity across organisations for generating and using evidence. This wide-ranging brief requires multifaceted approaches; assessing CLAHRCs’ success thus poses challenges for evaluation. This paper discusses these challenges in relation to seven CLAHRC evaluations, eliciting implications and suggestions for others evaluating similarly complex interventions with diverse objectives.
Cerium in human milk samples and its transfer from blood to milk: Is there an elevated nutritional risk for breast-fed babies?
Authors: Michalke Bernhard , González-Estecha Montserrat , Trasobares Elena M. , Herraiz Miguel Angel , Giussani Augusto , Oeh Uwe , Höllriegl Vera
Published: December 2011

The general population is increasingly exposed to cerium (Ce), which is contained in industrial products or is present as nuclear Ce fission products. Some studies suggested a link between elevated Ce levels and endomyocardial fibrosis. Since breast milk is the optimal, and directly after birth, usually the sole nutrition for newborns, exposure of females to Ce and its transfer to infants by breast-feeding is of concern in neonate protection. Consequently, the transfer rate of Ce from blood to breast milk is of interest for elucidating the Ce exposure of infants. Biomonitoring of paired serum and breast milk samples provides such information about Ce transfer to human milk. Therefore, this study is aimed at clarification of the relationship between Ce in human milk and serum from respective mothers for elucidating Ce enrichment in human milk with possible nutritional risk for newborns. As a prerequisite a strictly quality-controlled Ce determination method applicable to very low Ce concentration was developed, and its figures of merit were determined and found to be sufficient for our purpose. It turned out that Ce concentration in milk from Munich (Germany) and Madrid (Spain) showed a median of 13 ng/L. Ce concentrations in serum were at limit of quantification (LOQ) 10 ng/L (Munich) or 21.6–70.3 ng/L (Madrid), suggesting a higher Ce intake in Madrid. No enrichment from blood to milk was seen, and no elevated nutritional risk for breast-fed babies from Ce was found. Ce in serum, but not in milk, could indicate environmental Ce.
Syntheses and structures of two uranyl complexes with DMPU
Authors: Weis Eric M. , Duval Paul B. , Jurisson Silvia S.
Published: December 2011

Two uranyl complexes with the space-demanding ligand 1,3-dimethyltetrahydropyrimidin-2(1H)-one, DMPU, have been synthesized and characterized spectroscopically and by single crystal X-ray diffraction analysis.
Incorporation of pertechnetate and perrhenate into corroded steel surfaces studied by X-ray absorption fine structure spectroscopy
Authors: Heald S. M. , Krupka K. M. , Brown C. F.
Published: December 2011

Batch reaction experiments and solid-phase characterization analyses were completed to examine the uptake of dissolved perrhenate [Re(VII)] or pertechnetate [Tc(VII)] by A-516 steel coupons that corroded in simulated groundwater solutions or dilute water. The goal was to identify the mechanism(s) that control the uptake of99Tc by corrosion products on carbon steel in the presence of dilute solutions. X-ray absorption fine structure spectroscopy (XAFS) was used to study the oxidation states of Re and Tc incorporated into the corroded steel coupon surfaces. X-ray fluorescence maps showed that the corroded coupons contain localized regions enriched in Re or Tc. The ReL3near edge XAFS results for the coupons reacted with Re-spiked waters were consistent with nearly all of the sorbed Re being present as perrhenate and not significantly reduced to Re(IV). Linear combination fits of the extended XAFS signals for the perrhenate and ReIVO2standards indicate that Re sorbed to the steel coupons corroded in simulated J-13 (a relatively dilute Na-HCO3-CO3groundwater) and even more dilute waters consists of a maximum of 5 and 10% Re(IV), respectively. The fluorescence results also showed that the Re concentrations increased with increasing time of exposure to the X-ray beam, which suggests that the perrhenate ions are only weakly bonded to the matrix of the corrosion product. In contrast to the Re results, the TcKedge XAFS results for the coupons reacted in99Tc-spiked waters indicate that most of the sorbed Tc had been reduced to Tc(IV). The shape of the near edge and extended fine structure is similar to the Tc(IV)-hydrous ferric oxide (HFO) and not the TcO2·nH2O standard. Differences were noted in the XAFS results for steel coupons reacted with waters spiked with 0.001vs.0.1 mmol/L99Tc in that much more of the sorbed Tc from 0.001 mmol/L99Tc experiments was in the form of pertechnetate. Comparison of the XAFS results for coupons reacted with 0.001 mmol/L99Tc-spiked dilute simulated Na-HCO3-CO3groundwatervs.0.001 mmol/L99Tc-spiked dilute water also suggest that there are likely differences in the sorption mechanism for the pertechnetate fraction in the corrosion product which formed in these two test solutions. The cause for these differences is not known, but is likely due to differences in the compositions of the dilute simulated Na-HCO3-CO3groundwater and more dilute waters, such as the dissolved carbonate concentrations.
Interactions between oppositely-charged vs. those between like-charged species in crystal of aminoacetonitrile picrate: a charge density investigation
Authors: Nelyubina Yulia V. , Guru Row Tayur N. , Lyssenko Konstantin
Published: December 2011

Energy and charge aspects of two types of ion association – between oppositely-charged and between like-charged species – were quantified using the topological analysis of the electron density function derived from the low-temperature X-ray diffraction experiment for a crystal of aminoacetonitrile picrate (sp. gr. Cmca, Z = 8, R = 0.0187), providing an experimental evidence of their “equal rights” in crystal packing formation.
Child Pornography Sentencing and Demographic Data: Reforming Through Research
Authors: SpearIt
Published: December 2011

This article examines demographic research on child pornography offenders and considers its utility for sentencing reform. It begins by tracing the history of the Internet and federal possession law, detailing particularly how public and political panic about child pornography evolved within a growing fear of the Internet itself. The article continues by surveying current demographic research on possession offenders. Drawing on this data and related research, the article considers what the literature can contribute to sentencing policy, simultaneously showcasing vast differences between the type of offender Congress intended to punish and those actually receiving the harsh punishment. Taken wholly, this article explains why child pornography guidelines represent a departure from the normal process of creating Federal Sentencing Guidelines; it tells how law succumbed to the forces of fear and stacked the scale against child pornography offenders.
The molecular detection of relaxin and its receptor RXFP1 in reproductive tissue of Felis catus and Lynx pardinus during pregnancy
Authors: Braun Beate C , Vargas Astrid , Jewgenow Katarina
Published: December 2011

Relaxin acts as a pregnancy-specific signal in feline species, but specific information about protein structure and binding is essential for the improvement of pregnancy diagnosis in endangered feline species, like the Iberian lynx. To generate a felid-specific relaxin antibody, the DNA and protein sequences of lynx and cat were determined and peptides were chosen for antibody generation. In addition, relaxin and relaxin receptor (RXFP1) mRNA expressions were measured in uteri and ovaries of pregnant domestic cats and lynx placentae. Using real-time PCR and immunohistochemistry, it was established that feline placenta is the main source of relaxin during pregnancy. In other tested tissues, relaxin mRNA expression was weak. TheRXFP1mRNA expression was found mainly in cat uterine tissue and feline placentae. It was assumed that these tissues were main targets for relaxin. In the ovary, relaxin immunostaining was associated with blood vessels, signifying its role in vascularization.
Inhibition of germinal vesicle breakdown by antioxidants and the roles of signaling pathways related to nitric oxide and cGMP during meiotic resumption in oocytes of a marine worm
Authors: Stricker Stephen A
Published: December 2011

In mammalian oocytes, cAMP elevations prevent the resumption of meiotic maturation and thereby block nuclear disassembly (germinal vesicle breakdown (GVBD)), whereas nitric oxide (NO) and its downstream mediator cGMP can either inhibit or induce GVBD. Alternatively, some invertebrate oocytes use cAMP to stimulate, rather than inhibit, GVBD, and in such cases, the effects of NO/cGMP signaling on GVBD remain unknown. Moreover, potential interactions between NO/cGMP and AMP-activated kinase (AMPK) have not been assessed during GVBD. Thus, this study analyzed intraoocytic signaling pathways related to NO/cGMP in a marine nemertean worm that uses cAMP to induce GVBD. For such tests, follicle-free nemertean oocytes were stimulated to mature by seawater (SW) and cAMP elevators. Based on immunoblots and NO assays of maturing oocytes, SW triggered AMPK deactivation, NO synthase (NOS) phosphorylation, and an NO elevation. Accordingly, SW-induced GVBD was blocked by treatments involving the AMPK agonist AICAR, antioxidants, the NO scavenger carboxy-PTIO, NOS inhibitors, and cGMP antagonists that target the NO-stimulated enzyme, soluble guanylate cyclase (sGC). Conversely, SW solutions combining NO/cGMP antagonists with a cAMP elevator restored GVBD. Similarly, AICAR plus a cAMP-elevating drug reestablished GVBD while deactivating AMPK and phosphorylating NOS. Furthermore, sGC stimulators and 8-Br-cGMP triggered GVBD. Such novel results indicate that NO/cGMP signaling can upregulate SW-induced GVBD and that cAMP-elevating drugs restore GVBD by overriding the inhibition of various NO/cGMP downregulators, including AMPK. Moreover, considering the opposite effects of intraoocytic cAMP in nemerteans vs mammals, these data coincide with previous reports that NO/cGMP signaling blocks GVBD in rats.
Índice de manejo de carbono e atributos químicos de Latossolo Vermelho sob diferentes sistemas de manejo
Authors: Schiavo Jolimar Antonio , Rosset Jean Sérgio , Pereira Marcos Gervasio , Salton Júlio Cesar
Published: December 2011

O objetivo deste trabalho foi avaliar mudanças nos atributos químicos, no estoque de carbono e no índice de manejo de C de um Latossolo Vermelho argiloso, sob diferentes sistemas de manejo. Avaliou-se o solo sob sistemas de manejo com diferentes históricos de uso, sob as seguintes coberturas: Zea mays, Urochloa decumbens e Panicum maximum cv. Tanzania. Como testemunha, avaliou-se área sob vegetação de cerrado. Foram coletadas amostras compostas das camadas de solo de 0-5, 5-10 e 10-20 cm. Os teores de fósforo diminuíram com a profundidade, e os maiores valores foram observados na área com P. maximum. Na área com milho, observaram-se maiores valores de K+, Ca++, Mg++, CTC, e de soma e saturação por bases. O teor de C orgânico total do solo (COT) foi maior sob cerrado e menor sob cobertura com P. maximum, à profundidade de 5-10 cm. Os maiores estoques de COT, carbono nas frações particuladas (C-MOP) e nas frações associadas à fase mineral do solo ocorreram na área com U. decumbens, o que promoveu maiores índices de labilidade e de manejo de C, similares aos da vegetação de cerrado. Em todas as coberturas, a labilidade da matéria orgânica do solo diminuiu com o aumento da profundidade. A fração C-MOP apresenta maior sensibilidade às alterações de manejo e relaciona-se ao COT.
Desempenho e características das carcaças de novilhos superprecoces em pastos hibernais submetidos a intensidades de pastejo
Authors: Rocha Lemar Maciel da , Carvalho Paulo César de Faccio , Baggio Carolina , Anghinoni Ibanor , Lopes Marília Lazzarotto Terra , Macari Stefani , Silva Jamir Luis Silva da
Published: December 2011

O objetivo deste trabalho foi avaliar os efeitos de intensidades de pastejo na terminação de novilhos superprecoces, em pastagens de inverno em sucessão à soja. Foram avaliadas diferentes alturas de pré-pastejo de pastos mistos de azevém e aveia: 10, 20, 30 e 40 cm. Empregou-se o delineamento de blocos ao acaso, com três repetições. Foram utilizados machos mestiços de raças de corte, inteiros, com idade de 10 meses e peso médio inicial de 192±12 kg. As seguintes variáveis foram avaliadas: massa e oferta de forragem, altura do pasto, taxa de acúmulo diário e acúmulo total de forragem, ganho de peso médio diário (GMD), carga animal e ganho de peso vivo por unidade de área (GPA), além de características qualitativas de carcaça. Os novilhos foram abatidos aos 14 meses de idade. O GMD ajustou-se a um modelo de regressão quadrático e teve o ponto de máximo desempenho entre 20 e 25 cm de altura de pré-pastejo, enquanto o GPA ajustou-se a um modelo linear negativo. A qualidade da carcaça dos novilhos foi satisfatória, quando os pastos foram manejados em alturas superiores a 10 cm. É possível produzir carcaças de novilhos superprecoces, na fase de pastagem de sistemas integrados de produção, durante o inverno, em sucessão à soja. O manejo de pastos mistos de azevém e aveia, em alturas entre 20 e 25 cm, otimiza o desempenho animal individual e por área.