bluuebunny/crossref_metadata_2025 huggingface dataset:
CDK12 is a transcription elongation-associated CTD kinase, the metazoan ortholog of yeast Ctk1
Authors: Bartkowiak Bartlomiej , Liu Pengda , Phatnani Hemali P. , Fuda Nicholas J. , Cooper Jeffrey J. , Price David H. , Adelman Karen , Lis John T. , Greenleaf Arno L.
Published: October 2010

Drosophila contains one (dCDK12) and humans contain two (hCDK12 and hCDK13) proteins that are the closest evolutionary relatives of yeast Ctk1, the catalytic subunit of the major elongation-phase C-terminal repeat domain (CTD) kinase in Saccharomyces cerevisiae, CTDK-I. However, until now, neither CDK12 nor CDK13 has been demonstrated to be a bona fide CTD kinase. Using Drosophila, we demonstrate that dCDK12 (CG7597) is a transcription-associated CTD kinase, the ortholog of yCtk1. Fluorescence microscopy reveals that the distribution of dCDK12 on formaldehyde-fixed polytene chromosomes is virtually identical to that of hyperphosphorylated RNA polymerase II (RNAPII), but is distinct from that of P-TEFb (dCDK9 + dCyclin T). Chromatin immunoprecipitation (ChIP) experiments confirm that dCDK12 is present on the transcribed regions of active Drosophila genes. Compared with P-TEFb, dCDK12 amounts are lower at the 5′ end and higher in the middle and at the 3′ end of genes (both normalized to RNAPII). Appropriately, Drosophila dCDK12 purified from nuclear extracts manifests CTD kinase activity in vitro. Intriguingly, we find that cyclin K is associated with purified dCDK12, implicating it as the cyclin subunit of this CTD kinase. Most importantly, we demonstrate that RNAi knockdown of dCDK12 in S2 cells alters the phosphorylation state of the CTD, reducing its Ser2 phosphorylation levels. Similarly, in human HeLa cells, we show that hCDK13 purified from nuclear extracts displays CTD kinase activity in vitro, as anticipated. Also, we find that chimeric (yeast/human) versions of Ctk1 containing the kinase homology domains of hCDK12/13 (or hCDK9) are functional in yeast cells (and also in vitro); using this system, we show that a bur1ts mutant is rescued more efficiently by a hCDK9 chimera than by a hCDK13 chimera, suggesting the following orthology relationships: Bur1 ↔ CDK9 and Ctk1 ↔ CDK12/13. Finally, we show that siRNA knockdown of hCDK12 in HeLa cells results in alterations in the CTD phosphorylation state. Our findings demonstrate that metazoan CDK12 and CDK13 are CTD kinases, and that CDK12 is orthologous to yeast Ctk1.
CREB mediates brain serotonin regulation of bone mass through its expression in ventromedial hypothalamic neurons
Authors: Oury Franck , Yadav Vijay K. , Wang Ying , Zhou Bin , Liu X. Sherry , Guo X. Edward , Tecott Laurence H. , Schutz Günther , Means Anthony R. , Karsenty Gerard
Published: October 2010

Serotonin is a bioamine regulating bone mass accrual differently depending on its site of synthesis. It decreases accrual when synthesized in the gut, and increases it when synthesized in the brain. The signal transduction events elicited by gut-derived serotonin once it binds to the Htr1b receptor present on osteoblasts have been identified and culminate in cAMP response element-binding protein (CREB) regulation of osteoblast proliferation. In contrast, we do not know how brain-derived serotonin favors bone mass accrual following its binding to the Htr2c receptor on neurons of the hypothalamic ventromedial nucleus (VMH). We show here—through gene expression analysis, serotonin treatment of wild-type andHtr2c−/−hypothalamic explants, and cell-specific gene deletion in the mouse—that, following its binding to the Htr2c receptor on VMH neurons, serotonin uses a calmodulin kinase (CaMK)-dependent signaling cascade involving CaMKKβ and CaMKIV to decrease the sympathetic tone and increase bone mass accrual. We further show that the transcriptional mediator of these events is CREB, whose phosphorylation on Ser 133 is increased by CaMKIV following serotonin treatment of hypothalamic explants. A microarray experiment identified two genes necessary for optimum sympathetic activity whose expression is regulated by CREB. These results provide a molecular understanding of how serotonin signals in hypothalamic neurons to regulate bone mass accrual and identify CREB as a critical determinant of this function, although through different mechanisms depending on the cell type, neuron, or osteoblast in which it is expressed.
Interaction of noncoding RNA with the rDNA promoter mediates recruitment of DNMT3b and silencing of rRNA genes
Authors: Schmitz Kerstin-Maike , Mayer Christine , Postepska Anna , Grummt Ingrid
Published: October 2010

Noncoding RNAs are important components of regulatory networks controlling the epigenetic state of chromatin. We analyzed the role of pRNA (promoter-associated RNA), a noncoding RNA that is complementary to the rDNA promoter, in mediating de novo CpG methylation of rRNA genes (rDNA). We show that pRNA interacts with the target site of the transcription factor TTF-I, forming a DNA:RNA triplex that is specifically recognized by the DNA methyltransferase DNMT3b. The results reveal a compelling new mechanism of RNA-dependent DNA methylation, suggesting that recruitment of DNMT3b by DNA:RNA triplexes may be a common and generally used pathway in epigenetic regulation.
Microarray Analysis of the Abscission-Related Transcriptome in the Tomato Flower Abscission Zone in Response to Auxin Depletion
Authors: Meir Shimon , Philosoph-Hadas Sonia , Sundaresan Srivignesh , Selvaraj K.S. Vijay , Burd Shaul , Ophir Ron , Kochanek Bettina , Reid Michael S. , Jiang Cai-Zhong , Lers Amnon
Published: October 2010

The abscission process is initiated by changes in the auxin gradient across the abscission zone (AZ) and is triggered by ethylene. Although changes in gene expression have been correlated with the ethylene-mediated execution of abscission, there is almost no information on the molecular and biochemical basis of the increased AZ sensitivity to ethylene. We examined transcriptome changes in the tomato (Solanum lycopersicum ‘Shiran 1335’) flower AZ during the rapid acquisition of ethylene sensitivity following flower removal, which depletes the AZ from auxin, with or without preexposure to 1-methylcyclopropene or application of indole-3-acetic acid after flower removal. Microarray analysis using the Affymetrix Tomato GeneChip revealed changes in expression, occurring prior to and during pedicel abscission, of many genes with possible regulatory functions. They included a range of auxin- and ethylene-related transcription factors, other transcription factors and regulatory genes that are transiently induced early, 2 h after flower removal, and a set of novel AZ-specific genes. All gene expressions initiated by flower removal and leading to pedicel abscission were inhibited by indole-3-acetic acid application, while 1-methylcyclopropene pretreatment inhibited only the ethylene-induced expressions, including those induced by wound-associated ethylene signals. These results confirm our hypothesis that acquisition of ethylene sensitivity in the AZ is associated with altered expression of auxin-regulated genes resulting from auxin depletion. Our results shed light on the regulatory control of abscission at the molecular level and further expand our knowledge of auxin-ethylene cross talk during the initial controlling stages of the process.
Using rapid assessment and demographic methods to evaluate the effects of fishing on Heterodontus portusjacksoni off far‐eastern Victoria, Australia
Authors: Tovar‐Ávila J. , Day R. W. , Walker T. I.
Published: October 2010

A rapid semi‐quantitative ecological risk assessment method (productivity and susceptibility analysis) indicated that, despite its low biological productivity, the Port Jackson shark Heterodontus portusjacksoni is at low risk to all fishing methods in far‐eastern Victoria, Australia, under the present fishing practices, because of its low catch susceptibility. The risk to this population, however, would increase if the shark gillnet fishery operating in the region were to retain the species as a by‐product. Demographic analysis indicated that the species has medium intrinsic population growth rate and potential rebound in comparison with other chondrichthyan species, juveniles have higher elasticity than mature females and both juvenile and mature females have higher elasticities than hatchlings. Because of its low biological productivity and moderate resilience to the effects of fishing, cautious management measures will be necessary to ensure the sustainable use of H. portusjacksoni if its marketing increases in the future. Information on the dynamics of a population that is valuable to provide management advice can be obtained through demographic methods, but rapid assessment methods can also provide complementary information on the effects of fishing by considering the catch susceptibility of the population to each fishing method.
Rainbow trout Oncorhynchus mykiss consume less energy when swimming near obstructions
Authors: Cook C. L. , Coughlin D. J.
Published: October 2010

The effect of obstructions in steady flow on swimming by rainbow trout Oncorhynchus mykiss was examined in a respirometry swim tunnel to test the prediction that fish interacting with obstructions require less energy to hold station. When an obstruction was present, O. mykiss altered the kinematics of swimming and the rate of oxygen consumption was significantly reduced. The fish employed both entrainment and Kármán gait swimming strategies, permitting greater locomotor efficiency.
Glycoproteins histochemistry of the gills of Odontesthes bonariensis (Teleostei, Atherinopsidae)
Authors: Díaz A. O. , García A. M. , Escalante A. H. , Goldemberg A. L.
Published: October 2010

The histochemistry of glycoproteins (GP) in the mucous cells of the gills of the silverside Odontesthes bonariensis was identified with: (1) oxidizable vicinal diols; (2) sialic acid and some of their chain variants, carbon 7 (7C), carbon 8 (8C) or carbon 9 (9C); (3) sialic acid residues without O‐acyl substitution and with O‐acyl substitution at 7C, 8C or 9C; (4) carboxyl groups and (5) sulphate groups. A battery of seven biotinylated lectins allowed GPs sugar residues to be distinguished. Mucous cells showed the presence of neutral, sulphated and sialylated GPs. Dolichos biflorus agglutinin (DBA) and Glycine max agglutinin (SBA) showed strong positive staining; Arachis hypogaea agglutinin (PNA), Ricinus communis agglutinin‐I (RCA‐I) and Triticum vulgaris agglutinin (WGA) showed moderate staining, while Ulex europaeus agglutinin‐I (UEA‐I) was completely negative.
Systematic review: granulocyte/monocyte adsorptive apheresis for ulcerative colitis
Authors: Thanaraj S. , Hamlin P. J. , Ford A. C.
Published: October 2010

Aliment Pharmacol Ther 2010; 32: 1297–1306

Background  Patients with ulcerative colitis (UC) that is chronically active despite 5‐aminosalicylates or immunomodulators, or who are dependent on corticosteroids to maintain remission, have limited treatment options. Granulocyte/monocyte adsorptive apheresis (GMAA) may have a role in this situation.

Aim  To conduct a systematic review of GMAA in UC.

Methods  MEDLINE, EMBASE and the Cochrane central register of controlled trials were searched to identify randomized controlled trials (RCTs) comparing GMAA with conventional medical therapy, sham procedure or ‘intensive’ with ‘conventional’ GMAA regimens in adult UC patients. Studies reported clinical remission or response rates.

Results  Ten RCTs were eligible. Formal meta‐analysis was not undertaken due to concerns about methodological quality of identified studies. Compared with medical therapy, remission rates with GMAA were generally higher, and corticosteroid‐sparing effects were observed. Compared with sham procedure, GMAA did not achieve significantly higher remission rates. ‘Intensive’ GMAA regimens demonstrated generally higher remission rates, and time to remission was shorter compared with ‘conventional’ regimens. Only two RCTs were at low risk of bias. Six were conducted in Japanese patients, which may limit generalizability.

Conclusions  Granulocyte/monocyte adsorptive apheresis appears of some benefit in UC. High‐quality RCTs comparing granulocyte/monocyte adsorptive apheresis with conventional medical therapy or sham procedure in Western populations, with disease activity confirmed endoscopically, are required.
Effects of female feeding regime in a sexually cannibalistic mantid: fecundity, cannibalism, and male response inStagmomantis limbata(Mantodea)
Authors: MAXWELL MICHAEL R. , GALLEGO KEVIN M. , BARRY KATHERINE L.
Published: October 2010

1. Female feeding regime exhibited a cascade of effects on reproductive biology and behaviour in the mantidStagmomantis limbata(Hahn).

2. Well‐fed females (High diet) consistently attained greater body mass, thicker abdomens, and higher fecundity than food‐limited females (Low diet) in four annual generations. Dorso‐ventral abdominal thickness strongly correlated with fecundity.

3. In an experiment in captivity, Low diet females were more likely to cannibalize males than High diet females.

4. High diet females attracted more males than Low diet females in several contexts. In terms of long‐range attraction, caged High diet females attracted more males in the field. In terms of short‐range attraction, males preferentially mounted and copulated with High diet females in captive paired choice trials. In naturally‐occurring pairs in the field, a preference for females with thick abdomens was evident. These results point to two possible explanations: male choice for well‐fed females and state‐dependent female pheromone emission.

5. Two years of field data indicate considerable variation in female feeding success, as measured by abdominal thickness. Average feeding success in nature appears to be relatively high, when comparing the rate of abdominal expansion in the field to captive females maintained on High diets.

6. In the field, nearly all mounts and copulations occurred in September in both years, when female feeding success is expected to be high. If mating activity occurs during a window of high female feeding success, so as to reduce the risk of cannibalism for males, then the female feeding regime may be implicated in the phenology of mating activity in cannibalistic species.
Variability of larval Baltic sprat (Sprattus sprattus L.) otolith growth: a modeling approach combining spatially and temporally resolved biotic and abiotic environmental key variables
Authors: HINRICHSEN HANS‐HARALD , VOSS RUDI , HUWER BASTIAN , CLEMMESEN CATRIONA
Published: October 2010

Plankton sampling was conducted in the Baltic to obtain sprat larvae. Their individual drift patterns were back‐calculated using a hydrodynamic model. The modelled positions along the individual drift trajectories were subsequently used to provide insight into the environmental conditions experienced by the larvae. Autocorrelation analysis revealed that successive otolith increment widths of individual larvae were not independent. Otolith increment width was then modelled using two different generalized additive model (GAM) analyses (with and without autocorrelation), using environmental variables determined for each modelled individual larval position as explanatory variables. The results indicate that otolith growth was not only influenced by the density of potential prey but was controlled by a number of simultaneously acting environmental factors. The final model, not considering autocorrelation, explained more than 80% of the variance of otolith growth, with larval age as a factor variable showing the strongest significant impact on otolith growth. Otolith growth was further explained by statistically significant ambient environmental factors such as temperature, bottom depth, prey density and turbulence. The GAM analysis, taking autocorrelation into account, explained almost 98% of the variability, with the previous otolith increment showing the strongest significant effect. Larval age as well as ambient temperature and prey abundance also had a significant effect. An alternative approach applied individual‐based model (IBM) simulations on larval drift, feeding, growth and survival starting as exogenously feeding larvae at the back‐calculated positions. The IBM results revealed optimal growth conditions for more than 97% of the larvae, with a tendency for our IBM to slightly overestimate larval growth.
Influence of mesoscale eddies on ichthyoplankton assemblages in the Gulf of Alaska
Authors: ATWOOD ELIZABETH , DUFFY‐ANDERSON JANET T. , HORNE JOHN K. , LADD CAROL
Published: October 2010

Mesoscale eddies (100–200 km in diameter) propagating along the shelf‐break in the Gulf of Alaska are ubiquitous and have been shown to influence the ecosystem, but their influence on ichthyoplankton species composition and diversity has not been described. Evidence for larval fish entrainment in these eddies was examined using data from a cruise in 2005 that sampled three eastern Gulf of Alaska mesoscale eddies, and sampling that compared shelf to slope ichthyoplankton assemblages in the northern Gulf of Alaska (2002–2004). Hierarchical cluster analysis of oceanographic data showed that stations grouped according to location within an eddy. Species hierarchical cluster analysis revealed a latitudinal turnover in species composition, and an abundant species group. Species richness was correlated with distance from eddy center (P = 0.00025), and assemblages within eddies were significantly different (P < 0.05) from those in surrounding basin and shelf waters. These results suggest that mesoscale eddies propagating along the continental shelf‐break influence larval fish assemblages over the shelf and slope, which has implications for the timing and extent of larval fish distribution in the Gulf of Alaska.
Sudden cardiac death among people with diabetes: preventive measures documented in their medical records
Authors: Forslund Ann‐Sofie , Lundblad Dan , Söderberg Siv
Published: October 2010

Aims.  The purpose of this study was to examine how prevention of complications for people with diabetes mellitus had been conducted, as described in their medical records, focusing particularly on sudden cardiac death. A further aim was to compare the documentation with guidelines for diabetes care.

Background.  Diabetes mellitus is associated with an increased risk of cardiovascular disease, death and sudden cardiac death. About half of those affected by sudden cardiac death are assumed to have had one or more risk factors for cardiovascular disease that could have been treated effectively resulting in a reduced risk of sudden death.

Design.  Survey.

Method.  Fifty‐six people diagnosed with diabetes mellitus, who had died of a sudden cardiac arrest between the years 2003–2005, from the Northern Sweden MONICA myocardial registry were included. These people’s medical records were examined with regard to documentation of the care given during the year prior to the person’s sudden cardiac death.

Results.  The qualitative content analysis resulted in four categories: individualised goals for diabetes care; prevention of complications; self‐care; and factors which may affect ability to adhere to treatment. The quantitative analysis showed that few people with diabetes mellitus achieved goals for metabolic control, compared with those set in guidelines for diabetes mellitus care.

Conclusion.  To prevent complications for people with diabetes mellitus, it is a challenge for nurses and physicians to involve people with diabetes mellitus in their own care to improve the prognosis.

Relevance to clinical practice.  Examination of medical records of people with diabetes mellitus showed that documentation could be more informative and systematic. It is important to achieve better adherence to treatment and to increase people’s understanding of their illness.
The effectiveness of structured discharge education on maternal confidence, caring knowledge and growth of premature newborns
Authors: Shieh Shwu‐Jiuan , Chen Hsiu‐Lin , Liu Fen‐Chen , Liou Chuen‐Ching , Lin Y‐in‐Hui , Tseng Hsing‐I , Wang Ruey‐Hsia
Published: October 2010

Aims.  The aim of this study is to evaluate the effectiveness of structured discharge education on maternal confidence and caring knowledge and the growth of premature newborns.

Background.  Parents of premature newborns are usually confronted with great difficulties in caring for their babies after discharge. Building maternal confidence and caring knowledge can help mothers reduce such difficulties of caring for their babies after discharge from hospital.

Design.  Randomised controlled trial.

Methods.  Mothers with premature babies were randomly assigned into experimental (n = 29) and control groups (n = 30) at a medical centre in southern Taiwan. The mothers of both groups had received a questionnaire concerning maternal confidence and caring knowledge at pretest. After the pretest, a structured discharged education programme was provided to the mothers of the experimental group. The control group only received traditional discharge education. Mothers of experimental and control groups again received the questionnaire of maternal confidence and caring knowledge at the day before discharge and one month after discharge. At that time, the body height and body weight of newborns were measured and recorded.

Results.  Maternal confidence and caring knowledge of mothers in the experimental group were significantly higher than those of the control group at the day before discharge; however, there were no significant differences between the two groups one month after discharge. Using repeated anova analysis, the time × group interaction was significant for maternal confidence and caring knowledge, indicating that the groups differed significantly in changes in maternal confidence and caring knowledge over the three time points.

Conclusions.  Structured discharge education on mothers could significantly increase maternal confidence and caring knowledge at the day before discharge. Furthermore, structured discharge education could significantly increase the percentage of growth on body height of premature newborns.

Relevance to clinical practice.  The results could help clinical nurses design appropriate discharge education programmes for mothers of premature newborns.
A young clade repeating an old pattern: diversity inNothonotusdarters (Teleostei: Percidae) endemic to the Cumberland River
Authors: KECK BENJAMIN P. , NEAR THOMAS J.
Published: October 2010

Hypotheses of diversification in eastern North American freshwater fishes have focused primarily on allopatric distributions of species between disjunct highland areas and major river systems. However, these hypotheses do not fully explain the rich diversity of species within highland regions and river systems. Relatively old diversification events at small geographic scales have been observed in the Barcheek Darter subclade that occurs in the Cumberland River drainage (CRD) in Kentucky and Tennessee, United States of America, but it is unknown if this pattern is consistent in other darter subclades. We explored phylogeographic diversity in two species ofNothonotusdarters,N. microlepidusandN. sanguifluus, endemic to the CRD to compare phylogenetic patterns between Barcheek Darters and species ofNothonotus. We collected sequence data for a mitochondrial gene (cytb) and three nuclear genes (MLL, S7 and RAG1) from 19 N. microlepidusand 35 N. sanguifluusspecimens. Gene trees were estimated using maximum likelihood and Bayesian methods, and a ‘species tree’ was inferred using a Bayesian method. These trees indicate that species diversity inNothonotusis underestimated. Five distinct lineages were evident, despite retained ancestral polymorphism and unsampled extirpated populations. Comparison of chronograms for Barcheek Darters andNothonotusrevealed that microendemism resulting from species diversification at small geographic scales in the CRD is a consistent pattern in both old and young darter subclades. Our analyses reveal that geographic isolating mechanisms that result in similar phylogeographic patterns in the CRD are persistent through long expanses of evolutionary time.
Involvement of the Aspergillus nidulans protein kinase C with farnesol tolerance is related to the unfolded protein response
Authors: Colabardini Ana Cristina , de Castro Patrícia Alves , de Gouvêa Paula Fagundes , Savoldi Marcela , Malavazi Iran , Goldman Maria Helena S. , Goldman Gustavo Henrique
Published: October 2010

Previously, we demonstrated that the Aspergillus nidulans calC2 mutation in protein kinase C pkcA was able to confer tolerance to farnesol (FOH), an isoprenoid that has been shown to inhibit proliferation and induce apoptosis. Here, we investigate in more detail the role played by A. nidulans pkcA in FOH tolerance. We demonstrate that pkcA overexpression during FOH exposure causes increased cell death. FOH is also able to activate several markers of endoplasmic reticulum (ER) stress and the unfolded protein response (UPR). Our results suggest an intense cross‐talk between PkcA and the UPR during FOH‐induced cell death. Furthermore, the overexpression of pkcA increases both mRNA accumulation and metacaspases activity, and there is a genetic interaction between PkcA and the caspase‐like protein CasA. Mutant analyses imply that MAP kinases are involved in the signal transduction in response to the effects caused by FOH.
Review: Cerebral microvascular pathology in ageing and neurodegeneration
Authors: Brown W. R. , Thore C. R.
Published: October 2010

W. R. Brown and C. R. Thore (2011) Neuropathology and Applied Neurobiology37, 56–74 Cerebral microvascular pathology in ageing and neurodegeneration

This review of age‐related brain microvascular pathologies focuses on topics studied by this laboratory, including anatomy of the blood supply, tortuous vessels, venous collagenosis, capillary remnants, vascular density and microembolic brain injury. Our studies feature thick sections, large blocks embedded in celloidin, and vascular staining by alkaline phosphatase. This permits study of the vascular network in three dimensions, and the differentiation of afferent from efferent vessels. Current evidence suggests that there is decreased vascular density in ageing, Alzheimer's disease and leukoaraiosis, and cerebrovascular dysfunction precedes and accompanies cognitive dysfunction and neurodegeneration. A decline in cerebrovascular angiogenesis may inhibit recovery from hypoxia‐induced capillary loss. Cerebral blood flow is inhibited by tortuous arterioles and deposition of excessive collagen in veins and venules. Misery perfusion due to capillary loss appears to occur before cell loss in leukoaraiosis, and cerebral blood flow is also reduced in the normal‐appearing white matter. Hypoperfusion occurs early in Alzheimer's disease, inducing white matter lesions and correlating with dementia. In vascular dementia, cholinergic reductions are correlated with cognitive impairment, and cholinesterase inhibitors have some benefit. Most lipid microemboli from cardiac surgery pass through the brain in a few days, but some remain for weeks. They can cause what appears to be a type of vascular dementia years after surgery. Donepezil has shown some benefit. Emboli, such as clots, cholesterol crystals and microspheres can be extruded through the walls of cerebral vessels, but there is no evidence yet that lipid emboli undergo such extravasation.
Thermal tolerance of sympatric hymenopteran parasitoid species: does it match seasonal activity?
Authors: LE LANN CECILE , ROUX OLIVIER , SERAIN NELLY , VAN ALPHEN JACQUES J. M. , VERNON PHILIPPE , VAN BAAREN JOAN
Published: October 2010

Climatic changes result in an increased in mean temperature and in a higher incidence of extreme weather events such as heat and cold waves. For ectotherms, such as insect parasitoids, the ability to remain active under extreme climatic conditions is a significant key to fitness. The body size of individuals, and in particular their surface to volume ratio, may play a role in their resistance to thermal conditions. The thermal tolerances are investigated of two closely‐related sympatric parasitoid species [Aphidius avenae Haliday and Aphidius rhopalosiphi De Stefani‐Perez (Hymenoptera: Aphidiinae)] that have a similar ecology but differ in body size and phenologies. The critical thermal limits of individuals are assessed in both sexes of each parasitoid species and the influence of surface–volume ratios on their thermal tolerances. Aphidius avenae is less resistant to low temperatures and more resistant to high temperatures than A. rhopalosiphi. The lower surface to volume ratio of A. avenae individuals may help them to remain active in summer when experiencing heat waves. However, body size is not the sole factor that plays a role in differences of thermal tolerance between species and body size may not be an adaptation to extreme temperatures but rather a by‐product of developmental regulation. Closely‐related sympatric species from the same ecological guild can have different thermal tolerances that may allow them to occur within the same habitat. The present study also highlights the importance of clearly defining how to measure critical thermal limits to determine the thermal tolerance of a species.
Cryptogein, a fungal elicitor, remodels the phenylpropanoid metabolism of tobacco cell suspension cultures in a calcium‐dependent manner
Authors: AMELOT NICOLAS , CARROUCHE AUDREY , DANOUN SAÏDA , BOURQUE STÉPHANE , HAIECH JACQUES , PUGIN ALAIN , RANJEVA RAOUL , GRIMA‐PETTENATI JACQUELINE , MAZARS CHRISTIAN , BRIERE CHRISTIAN
Published: October 2010

Plant cells use calcium‐based signalling pathways to transduce biotic and/or abiotic stimuli into adaptive responses. However, little is known about the coupling between calcium signalling, transcriptional regulation and the downstream biochemical processes. To understand these relationships better, we challenged tobacco BY‐2 cells with cryptogein and evaluated how calcium transients (monitored through the calcium sensor aequorin) impact (1) transcript levels of phenylpropanoid genes (assessed by RT‐qPCR); and (2) derived‐phenolic compounds (analysed by mass spectrometry). Most genes of the phenylpropanoid pathway were up‐regulated by cryptogein and cell wall‐bound phenolic compounds accumulated (mainly 5‐hydroxyferulic acid). The accumulation of both transcripts and phenolics was calcium‐dependent. The transcriptional regulation of phenylpropanoid genes was correlated in a non‐linear manner with stimulus intensity and with components of the cryptogein‐induced calcium signature. In addition, calmodulin inhibitors increased the sensitivity of cells to low concentrations of cryptogein. These results led us to propose a model of coupling between the cryptogein signal, calcium signalling and the transcriptional response, exerting control of transcription through the coordinated action of two decoding modules exerting opposite effects.
Biochemical, molecular and preclinical characterization of a double‐virus‐reduced human butyrylcholinesterase preparation designed for clinical use
Authors: Weber A. , Butterweck H. , Mais‐Paul U. , Teschner W. , Lei L. , Muchitsch E.‐M. , Kolarich D. , Altmann F. , Ehrlich H. J. , Schwarz H. P.
Published: October 2010

Background and Objectives A human plasma‐derived butyrylcholinesterase preparation manufactured on the industrial scale is described.

Material and Methods The human butyrylcholinesterase (hBChE) product was extensively investigated for its purity using immunological and electrophoretic methods and characterized by thorough glycoproteomic approaches. A comprehensive preclinical testing programme addressing safety and pharmacokinetic parameters supplemented the biochemical characterization.

Results The high‐purity hBChE preparation is tetrameric and has high specific activity and molecular integrity of the protein backbone. Acute toxicity studies andin vivothrombogenicity studies provided evidence of a sufficient safety margin for use in humans.

Conclusion Extensive preclinical safety and pharmacokinetic testing confirmed that this hBChE preparation can be used for further efficacy testing as a bioscavenger for toxic organophosphate compounds in appropriate animal models and ultimately in humans.
Population genetic structure of the endangered limpetCymbula nigrain a temperate Northern hemisphere region: influence of palaeoclimatic events?
Authors: Espinosa Free , Nakano Tomoyuki , Guerra‐García Jose M. , García‐Gómez Jose C.
Published: October 2010

Cymbula nigra(da Costa L, 1771) is the largest limpet in Europe and it is at present considered an endangered species. Nevertheless, its biology is poorly understood and there is a lack of knowledge about population genetic structure. A total of 61C. nigraspecimens were sampled and studied for a fragment of the mitochondrial gene cytochromecoxidase subunit I. The low genetic diversity, the absence of geographic structure, the significant values reported by measures to test for a recent population expansion and a star‐shaped phylogeny pattern are here interpreted as forming a hypothesis concerning the influence of palaeoclimatic events in the genetic pool of limpets, considering also the information available on different limpet species.
In Vitro Fertilization using Non‐sexed and Sexed Bovine Sperm: Sperm Concentration, Sorter Pressure, and Bull Effects
Authors: Barceló‐Fimbres M , Campos‐Chillón LF , Seidel Jr GE
Published: October 2010

Contents

The objective of these experiments was to study bovine in vitro fertilization (IVF) conditions for blastocyst production using non‐sexed sperm (Experiment 1) and sexed sperm (Experiment 2). For Experiment 1, in vitro‐matured oocytes (N = 707) were allocated to a 2 × 3 × 4 factorial design: time of co‐incubation of gametes for fertilization (4 and 18 h), sperm dose (1, 0.33, and 0.11 × 106 frozen–thawed sperm/ml, and sperm source (four bulls). Pronuclear status was evaluated for a subset. Experiment 2 (N = 2155 oocytes) was a 2 × 3 × 2 × 6 factorial design: sex of sperm (X and Y), sperm dose (1, 0.33, and 0.11 × 106 frozen–thawed sperm/ml), and sperm‐sorting pressures (40 and 50 psi), replicated with sperm of six bulls. Presumptive zygotes were cultured 60 h in chemically defined medium‐1 (CDM‐1), and for 114 h in CDM‐2. For Experiment 1, pronuclear formation, cleavage and blastocysts rates were greater for 1, and 0.33 × 106 than 0.11 × 106 sperm/ml (72 and 62 vs 42%; 89 and 81 vs 58%; and 21 and 17 vs 9%, respectively; all p < 0.01); polyspermy was greater for 1, than 0.33 and 0.11 × 106 sperm concentrations (24 vs 2 and 0%; p < 0.01). There were greater main effects (p < 0.01) of pronuclear formation (69 vs 48%), polyspermy (13 vs 4%), and cleavage (63 vs 54%), at 18 than at 4 h of co‐incubation of gametes (all p < 0.01). For Experiment 2, cleavage and blastocyst rates were greater for 1 × 106 sperm/ml vs 0.33 and 0.11 (69%, 47%, and 30% cleavage and 30%, 14%, and 8% blastocysts) and 40 vs 50 psi (54% and 44% cleavage and 18% and 15% blastocysts) (p < 0.01). A marked bull by fertilization sperm dose interaction was found for cleavage (p < 0.05). The main conclusion was that the optimal sperm concentration for cleavage and producing blastocysts via IVF with sexed sperm was considerably higher and more variable among bulls than for unsexed sperm.
Two cases of Reciprocal Chromosomal Translocation (4; 7)(p+; q−) (2; 8)(q−; q+) in Piglets Produced by ICSI
Authors: García‐Vázquez FA , Hernández‐Caravaca I , Martín M , Gómez E , Rodríguez A , Sánchez‐Sánchez R , Gadea J
Published: October 2010

Contents

In this study, the karyotypes of 14 piglets from four different litters produced by intracytoplasmic sperm injection (ICSI) and embryo transfer were analysed. The chromosome analysis was based on a classical cytogenetic examination following the standard protocols of lymphocyte cultures. Two cases of reciprocal translocation [(4; 7)(p+; q−) and (2; 8)(q−; q+)] were detected in two female transgenic piglets. These animals showed neither anatomical nor physiological alterations and had normal growth. To our knowledge, this is the first karyotype study of piglets produced by ICSI.
Preeclamptic nephropathy
Authors: HENNESSY ANNEMARIE , MAKRIS ANGELA
Published: October 2010

With the recent discovery of potential serum ‘toxins’ in human preeclampsia, it is timely to consider how these might relate to preeclamptic nephropathy. This review will discuss the clinical presentation of preeclampsia with an emphasis on renal involvement. It will explore the nature of the renal histological changes including endothelial and the more recently discovered podocyte changes, the implications of elevated anti‐angiogenic molecules, anti‐angiotensin‐1 receptor agonistic antibodies and other proposed mechanisms in causing or exacerbating renal lesions. It will also explore the role of pre‐existing renal disease in causing preeclampsia and the potential for new biomarkers, both serum and urinary, to inform clinical practice with regard to differentiating preeclampsia from pre‐existing renal disease. Recommendations about the future of women who have had preeclampsia are unclear but the general consensus is that there are future cardiovascular risks, and to a lesser extent, future renal risks in these women. Regular review of proteinuria and glomerular filtration rate as well as overall cardiovascular risk status seems a logical step.
Understanding cardiac biomarkers in end‐stage kidney disease: Frequently asked questions and the promise of clinical application
Authors: ROBERTS MATTHEW A , HEDLEY ADAM J , IERINO FRANCESCO L
Published: October 2010

A novel strategy in the management of cardiovascular disease in patients with end‐stage kidney disease is the use of biochemical markers to facilitate the detection of cardiovascular abnormalities in the hope that this will allow effective therapy to be instituted earlier. The cardiac troponins and B‐type natriuretic peptide are among the best studied of these biochemical markers of cardiovascular disease. However, controversy remains regarding the interpretation of such results and the subsequent clinical application of these biomarkers, particularly when abnormal in patients with end‐stage kidney disease. This review addresses some of the important issues to consider with the interpretation of abnormal cardiac troponin and B‐type natriuretic peptide results in patients undergoing dialysis.
Carvedilol protects tubular epithelial cells from ischemia–reperfusion injury by inhibiting oxidative stress
Authors: Hayashi Taiji , De Velasco Marco Antonio , Saitou Yoshitaka , Nose Kazuhiro , Nishioka Tsukasa , Ishii Tokumi , Uemura Hirotsugu
Published: October 2010

Objectives: Renal ischemia–reperfusion injury (IRI), leading to acute kidney injury, is a frequent complication with renal transplantation and it is associated with graft function. Its pathogenesis involves ischemia, vascular congestion and reactive oxygen metabolites. Carvedilol is an antihypertensive drug with potent anti‐oxidant properties. In this study we investigated the protective effects of carvedilol in a rat renal IRI model.

Methods: Twenty‐four rats were randomized into sham, untreated control and carvedilol (2 mg/kg 30 min before surgery and 12 hr after reperfusion) treatment groups and were subjected to 60 min of left renal ischemia followed by reperfusion at 24, 48, 96 and 168 hr.

Results: Treatment with carvedilol significantly decreased plasma creatinine levels after IRI (up to 168 hr) compared to controls (P < 0.001), suggesting an improvement in renal function. Histopathological analysis revealed decreased IRI‐induced damage in kidneys from carvedilol‐treated rats. A significant increase in the expression levels of Cu/Zn superoxide dismutase and reduction of 8‐hydroxydeoxyguanosine and apoptosis levels (P < 0.005) suggested a protective effect after treatment with carvedilol.

Conclusions: Our findings suggest that carvedilol ameliorates IRI resulting in improved renal function.
Retinoic acid receptor‐α signalling antagonizes both intracellular and extracellular amyloid‐β production and prevents neuronal cell death caused by amyloid‐β
Authors: Jarvis C. I. , Goncalves M. B. , Clarke E. , Dogruel M. , Kalindjian S. B. , Thomas S. A. , Maden M. , Corcoran J. P. T.
Published: October 2010

Alzheimer’s disease (AD) is characterized by amyloid‐β (Aβ) deposition in the brain, neuronal cell loss and cognitive decline. We show here that retinoic acid receptor (RAR)α signalling in vitro can prevent both intracellular and extracellular Aβ accumulation. RARα signalling increases the expression of a disintegrin and metalloprotease 10, an α‐secretase that processes the amyloid precursor protein into the non‐amyloidic pathway, thus reducing Aβ production. We also show that RARα agonists are neuroprotective, as they prevent Aβ‐induced neuronal cell death in cortical cultures. If RARα agonists are given to the Tg2576 mouse, the normal Aβ production in their brains is suppressed. In contrast, neither RARβ nor γ‐agonists affect Aβ production or Aβ‐mediated neuronal cell death. Therefore, RARα agonists have therapeutic potential for the treatment of AD.
Representations of visual proportions in the primate posterior parietal and prefrontal cortices
Authors: Vallentin Daniela , Nieder Andreas
Published: October 2010

The primate prefrontal (PFC) and posterior parietal cortices (PPC) have been shown to be cardinal structures in processing abstract absolute magnitudes, such as numerosity or length. The neuronal representation of quantity relations, however, remained largely elusive. Recent functional imaging studies in humans showed that blood flow changes systematically both in the PFC and the PPC as a function of relational distance between proportions. We investigated the response properties of single neurons in the lateral PFC and the inferior parietal lobule (IPL, area 7) in rhesus monkeys performing a lengths‐proportion‐discrimination task. Neurons in both areas shared many characteristics and showed peaked tuning functions with preferred proportions. However, a significantly higher percentage of neurons coding proportions was found in the PFC compared with the IPL. In agreement with human studies, our study shows that proportions are represented in the fronto‐parietal network that has already been implicated for absolute magnitude processing.
Deglycosylation of puerarin and other aromatic C‐glucosides by a newly isolated human intestinal bacterium
Authors: Braune Annett , Blaut Michael
Published: October 2010

The human intestinal microbiota may influence the fate of bioactive polyphenols, such as the isoflavone puerarin (daidzein 8‐C‐glucoside), following their oral intake. Faecal suspensions from 19 healthy subjects were tested for their ability to C‐deglycosylate puerarin. Only one of these catalysed this reaction. A rod‐shaped Gram‐positive bacterium, strain CG19‐1, capable of deglycosylating puerarin to daidzein was isolated from the corresponding suspension. However, the strictly anaerobic isolate was unable to utilize puerarin as sole carbon and energy source nor any of the tested carbohydrates. Comparative 16S rRNA gene sequence analysis indicated that strain CG19‐1 is a new species of the Lachnospiraceae. Strain CG19‐1 also converted other aromatic C‐glucosides in addition to puerarin. The xanthone C‐glucoside mangiferin was deglycosylated to norathyriol. The flavone C‐glucosides homoorientin and vitexin were degraded to 3‐(3,4‐dihydroxyphenyl)propionic acid via luteolin and 3‐(4‐hydroxyphenyl)propionic acid respectively. In addition, strain CG19‐1 converted flavonoid O‐glucosides, but at rates that were lower than those of the C‐glucosides tested. The isolate deglycosylated the isoflavone O‐glucosides daidzin and genistin to daidzein and genistein respectively. Several O‐glucosides of the flavones luteolin and apigenin undergoing deglycosylation were subsequently cleaved to 3‐(3,4‐dihydroxyphenyl)propionic acid and 3‐(4‐hydroxyphenyl)propionic acid respectively. Moreover, strain CG19‐1 cleaved both O‐desmethylangolensin and 6′‐hydroxy‐O‐desmethylangolensin to yield 2‐(4‐dihydroxyphenyl)propionic acid. The corresponding cleavage product, resorcinol, was only observed for O‐desmethylangolensin.
A novel, mat‐forming Thiomargarita population associated with a sulfidic fluid flow from a deep‐sea mud volcano
Authors: Girnth Anne‐Christin , Grünke Stefanie , Lichtschlag Anna , Felden Janine , Knittel Katrin , Wenzhöfer Frank , de Beer Dirk , Boetius Antje
Published: October 2010

A mat‐forming population of the giant sulfur bacterium Thiomargarita was discovered at the flank of the mud volcano Amon on the Nile Deep Sea Fan in the Eastern Mediterranean Sea. All cells were of a spherical and vacuolated phenotype and internally stored globules of elemental sulfur. With a diameter of 24–65 µm, Thiomargarita cells from the Eastern Mediterranean were substantially smaller than cells of previously described populations. A 16S rRNA gene fragment was amplified and could be assigned to the Thiomargarita‐resembling cells by fluorescence in situ hybridization. This sequence is monophyletic with published Thiomargarita sequences but sequence similarities are only about 94%, indicating a distinct diversification. In the investigated habitat, highly dynamic conditions favour Thiomargarita species over other sulfur‐oxidizing bacteria. In contrast to Thiomargarita namibiensis populations, which rely on periodic resuspension from sulfidic sediment into the oxygenated water column, Thiomargarita cells at the Amon mud volcano seem to remain stationary at the sediment surface while environmental conditions change around them due to periodic brine flow.
A genetic element present on megaplasmids allows Enterococcus faecium to use raffinose as carbon source
Authors: Zhang Xinglin , Vrijenhoek Joyce E. P. , Bonten Marc J. M. , Willems Rob J. L. , van Schaik Willem
Published: October 2010

Enterococcus faecium is a commensal of the gastrointestinal tract of humans and animals. Since the 1990s, it has also emerged as a nosocomial pathogen. Little is known about carbon metabolism of E. faecium even though the ability to utilize different sugars could be an important factor in adapting to different ecological niches. In this study we identify an E. faecium gene cluster that is responsible for the metabolism of the α‐galactoside sugar raffinose. Phenotypic testing of seven E. faecium isolates of which the genomes were previously sequenced showed that one isolate (strain E980) could grow on raffinose. Genome analysis identified a gene cluster containing two genes encoding α‐galactosidases (termed agaA and agaB) that was uniquely present in E980. The agaA and agaB genes were significantly more frequently found in strains that are phylogenetically related to E980 and were more prevalent in surveillance isolates from hospital and community sources than in isolates from clinical infections. Disruption of the α‐galactosidase gene agaB, but not of agaA, disabled growth on raffinose in strain E980. In all strains agaA and agaB are carried on megaplasmids that are between 150 and 300 kb in size. Filter‐mating experiments showed that the megaplasmid of E980 can be transferred to a plasmidless recipient which then gains the ability to grow on raffinose. The observation that raffinose utilization by E. faecium is a trait carried by megaplasmids indicates that these megaplasmids can have important roles in shaping the competitive fitness of E. faecium in the environment, for example by expanding the metabolic repertoire of this organism.