bluuebunny/crossref_metadata_2025 huggingface dataset:
Deep Sequencing revealed ‘Plant Like Transcripts’ in mosquitoAnopheles culicifacies: an Evolutionary Puzzle
Authors: Sharma Punita , Sharma Swati , Mishra Ashwani Kumar , Thomas Tina , Das De Tanwee , Verma Sonia , Kumari Vandana , Rohilla Suman Lata , Singh Namita , Pandey Kailash C , Dixit Rajnikant
Published: October 2014

As adult female mosquito’s salivary gland facilitate blood meal uptake and pathogen transmission e.g.Plasmodium, virus etc., a plethora of research has been focused to understand the mosquito-vertebrate-pathogen interactions. Despite the fact that mosquito spends longer time over nectar sugar source, the fundamental question ‘how adult female salivary gland’ manages molecular and functional relationship during sugar vs. blood meal uptake remains unanswered. Currently, we are trying to understand these molecular relationships under dual feeding conditions in the salivary glands of the mosquitoAnopheles culicifacies. During functional annotation of salivary transcriptome database, unexpectedly we discovered a cluster of salivary transcripts encoding plant like proteins. Our multiple experimental validations confirmed that Plant like transcripts (PLTs) are of mosquito origin and may encode functional proteins. A comprehensive molecular analysis of the PLTs and ongoing metagenomic analysis of salivary microbiome provide first evidence that how mosquito may have been benefited from its association with plant host and microbes. Future understanding of the underlying mechanism of the feeding associated molecular responses may provide new opportunity to control vector borne diseases.
Short intronic repeat sequences facilitate circular RNA production
Authors: Liang Dongming , Wilusz Jeremy E.
Published: October 2014

Recent deep sequencing studies have revealed thousands of circular noncoding RNAs generated from protein-coding genes. These RNAs are produced when the precursor messenger RNA (pre-mRNA) splicing machinery “backsplices” and covalently joins, for example, the two ends of a single exon. However, the mechanism by which the spliceosome selects only certain exons to circularize is largely unknown. Using extensive mutagenesis of expression plasmids, we show that miniature introns containing the splice sites along with short (∼30- to 40-nucleotide) inverted repeats, such as Alu elements, are sufficient to allow the intervening exons to circularize in cells. The intronic repeats must base-pair to one another, thereby bringing the splice sites into close proximity to each other. More than simple thermodynamics is clearly at play, however, as not all repeats support circularization, and increasing the stability of the hairpin between the repeats can sometimes inhibit circular RNA biogenesis. The intronic repeats and exonic sequences must collaborate with one another, and a functional 3′ end processing signal is required, suggesting that circularization may occur post-transcriptionally. These results suggest detailed and generalizable models that explain how the splicing machinery determines whether to produce a circular noncoding RNA or a linear mRNA.
PINOID AGC Kinases Are Necessary for Phytochrome-Mediated Enhancement of Hypocotyl Phototropism in Arabidopsis
Authors: Haga Ken , Hayashi Ken-ichiro , Sakai Tatsuya
Published: October 2014

Several members of the AGCVIII kinase subfamily, which includes PINOID (PID), PID2, and WAVY ROOT GROWTH (WAG) proteins, have previously been shown to phosphorylate PIN-FORMED (PIN) auxin transporters and control the auxin flow in plants. PID has been proposed as a key component of the phototropin signaling pathway that induces phototropic responses, although the responses were not significantly impaired in the pid single and pid wag1 wag2 triple mutants. This raises questions about the functional roles of the PID family in phototropic responses. Here, we investigated hypocotyl phototropism in the pid pid2 wag1 wag2 quadruple mutant in detail to clarify the roles of the PID family in Arabidopsis (Arabidopsis thaliana). The pid quadruple mutants exhibited moderate responses in continuous light-induced phototropism with a decrease in growth rates of hypocotyls and normal responses in pulse-induced phototropism. However, they showed serious defects in enhancements of pulse-induced phototropic curvatures and lateral fluorescent auxin transport by red light pretreatment. Red light pretreatment significantly reduced the expression level of PID, and the constitutive expression of PID prevented pulse-induced phototropism, irrespective of red light pretreatment. This suggests that the PID family plays a significant role in phytochrome-mediated phototropic enhancement but not the phototropin signaling pathway. Red light treatment enhanced the intracellular accumulation of PIN proteins in response to the vesicle-trafficking inhibitor brefeldin A in addition to increasing their expression levels. Taken together, these results suggest that red light preirradiation enhances phototropic curvatures by up-regulation of PIN proteins, which are not being phosphorylated by the PID family.
Arabidopsis PHOSPHOTYROSYL PHOSPHATASE ACTIVATOR Is Essential for PROTEIN PHOSPHATASE 2A Holoenzyme Assembly and Plays Important Roles in Hormone Signaling, Salt Stress Response, and Plant Development
Authors: Chen Jian , Hu Rongbin , Zhu Yinfeng , Shen Guoxin , Zhang Hong
Published: October 2014

PROTEIN PHOSPHATASE 2A (PP2A) is a major group of serine/threonine protein phosphatases in eukaryotes. It is composed of three subunits: scaffolding subunit A, regulatory subunit B, and catalytic subunit C. Assembly of the PP2A holoenzyme in Arabidopsis (Arabidopsis thaliana) depends on Arabidopsis PHOSPHOTYROSYL PHOSPHATASE ACTIVATOR (AtPTPA). Reduced expression of AtPTPA leads to severe defects in plant development, altered responses to abscisic acid, ethylene, and sodium chloride, and decreased PP2A activity. In particular, AtPTPA deficiency leads to decreased methylation in PP2A-C subunits (PP2Ac). Complete loss of PP2Ac methylation in the suppressor of brassinosteroid insensitive1 mutant leads to 30% reduction of PP2A activity, suggesting that PP2A with a methylated C subunit is more active than PP2A with an unmethylated C subunit. Like AtPTPA, PP2A-A subunits are also required for PP2Ac methylation. The interaction between AtPTPA and PP2Ac is A subunit dependent. In addition, AtPTPA deficiency leads to reduced interactions of B subunits with C subunits, resulting in reduced functional PP2A holoenzyme formation. Thus, AtPTPA is a critical factor for committing the subunit A/subunit C dimer toward PP2A heterotrimer formation.
Differentially Phased Leaf Growth and Movements in Arabidopsis Depend on Coordinated Circadian and Light Regulation
Authors: Dornbusch Tino , Michaud Olivier , Xenarios Ioannis , Fankhauser Christian
Published: October 2014

In contrast to vastly studied hypocotyl growth, little is known about diel regulation of leaf growth and its coordination with movements such as changes in leaf elevation angle (hyponasty). We developed a 3D live-leaf growth analysis system enabling simultaneous monitoring of growth and movements. Leaf growth is maximal several hours after dawn, requires light, and is regulated by daylength, suggesting coupling between growth and metabolism. We identify both blade and petiole positioning as important components of leaf movements in Arabidopsis thaliana and reveal a temporal delay between growth and movements. In hypocotyls, the combination of circadian expression of PHYTOCHROME INTERACTING FACTOR4 (PIF4) and PIF5 and their light-regulated protein stability drives rhythmic hypocotyl elongation with peak growth at dawn. We find that PIF4 and PIF5 are not essential to sustain rhythmic leaf growth but influence their amplitude. Furthermore, EARLY FLOWERING3, a member of the evening complex (EC), is required to maintain the correct phase between growth and movement. Our study shows that the mechanisms underlying rhythmic hypocotyl and leaf growth differ. Moreover, we reveal the temporal relationship between leaf elongation and movements and demonstrate the importance of the EC for the coordination of these phenotypic traits.
Critical Function of a Chlamydomonas reinhardtii Putative Polyphosphate Polymerase Subunit during Nutrient Deprivation
Authors: Aksoy Munevver , Pootakham Wirulda , Grossman Arthur R.
Published: October 2014

Forward genetics was used to isolate Chlamydomonas reinhardtii mutants with altered abilities to acclimate to sulfur (S) deficiency. The ars76 mutant has a deletion that eliminates several genes, including VACUOLAR TRANSPORTER CHAPERONE1 (VTC1), which encodes a component of a polyphosphate polymerase complex. The ars76 mutant cannot accumulate arylsulfatase protein or mRNA and shows marked alterations in levels of many transcripts encoded by genes induced during S deprivation. The mutant also shows little acidocalcisome formation compared with wild-type, S-deprived cells and dies more rapidly than wild-type cells following exposure to S-, phosphorus-, or nitrogen (N)-deficient conditions. Furthermore, the mutant does not accumulate periplasmic l-amino acid oxidase during N deprivation. Introduction of the VTC1 gene specifically complements the ars76 phenotypes, suggesting that normal acidocalcisome formation in cells deprived of S requires VTC1. Our data also indicate that a deficiency in acidocalcisome function impacts trafficking of periplasmic proteins, which can then feed back on the transcription of the genes encoding these proteins. These results and the reported function of vacuoles in degradation processes suggest a major role of the acidocalcisome in reshaping the cell during acclimation to changing environmental conditions.
ArabidopsisATG8-INTERACTING PROTEIN1 Is Involved in Autophagy-Dependent Vesicular Trafficking of Plastid Proteins to the Vacuole
Authors: Michaeli Simon , Honig Arik , Levanony Hanna , Peled-Zehavi Hadas , Galili Gad
Published: October 2014

Selective autophagy has been extensively studied in various organisms, but knowledge regarding its functions in plants, particularly in organelle turnover, is limited. We have recently discovered ATG8-INTERACTING PROTEIN1 (ATI1) from Arabidopsis thaliana and showed that following carbon starvation it is localized on endoplasmic reticulum (ER)-associated bodies that are subsequently transported to the vacuole. Here, we show that following carbon starvation ATI1 is also located on bodies associating with plastids, which are distinct from the ER ATI bodies and are detected mainly in senescing cells that exhibit plastid degradation. Additionally, these plastid-localized bodies contain a stroma protein marker as cargo and were observed budding and detaching from plastids. ATI1 interacts with plastid-localized proteins and was further shown to be required for the turnover of one of them, as a representative. ATI1 on the plastid bodies also interacts with ATG8f, which apparently leads to the targeting of the plastid bodies to the vacuole by a process that requires functional autophagy. Finally, we show that ATI1 is involved in Arabidopsis salt stress tolerance. Taken together, our results implicate ATI1 in autophagic plastid-to-vacuole trafficking through its ability to interact with both plastid proteins and ATG8 of the core autophagy machinery.
Chromatin-Dependent Repression of theArabidopsisFloral Integrator Genes Involves Plant Specific PHD-Containing Proteins
Authors: López-González Leticia , Mouriz Alfonso , Narro-Diego Laura , Bustos Regla , Martínez-Zapater José Miguel , Jarillo Jose A. , Piñeiro Manuel
Published: October 2014

The interplay among histone modifications modulates the expression of master regulatory genes in development. Chromatin effector proteins bind histone modifications and translate the epigenetic status into gene expression patterns that control development. Here, we show that two Arabidopsis thaliana paralogs encoding plant-specific proteins with a plant homeodomain (PHD) motif, SHORT LIFE (SHL) and EARLY BOLTING IN SHORT DAYS (EBS), function in the chromatin-mediated repression of floral initiation and play independent roles in the control of genes regulating flowering. Previous results showed that repression of the floral integrator FLOWERING LOCUS T (FT) requires EBS. We establish that SHL is necessary to negatively regulate the expression of SUPPRESSOR OF OVEREXPRESSION OF CO1 (SOC1), another floral integrator. SHL and EBS recognize di- and trimethylated histone H3 at lysine 4 and bind regulatory regions of SOC1 and FT, respectively. These PHD proteins maintain an inactive chromatin conformation in SOC1 and FT by preventing high levels of H3 acetylation, bind HISTONE DEACETYLASE6, and play a central role in regulating flowering time. SHL and EBS are widely conserved in plants but are absent in other eukaryotes, suggesting that the regulatory module mediated by these proteins could represent a distinct mechanism for gene expression control in plants.
Genomic characterization of ϕRS603, a filamentous bacteriophage that is infectious to the phytopathogen Ralstonia solanacearum
Authors: Van Truong Thi Bich , Yoshida Shohei , Miki Kaito , Kondo Akihiro , Kamei Kaeko
Published: October 2014

A filamentous bacteriophage (ϕ), ϕRS603, which is infectious to the phytopathogen Ralstonia solanacearum was isolated. ϕRS603 was found to have a circular single‐stranded DNA genome composed of 7679 nucleotides and to contain 13 putative open reading frames (ORFs). The ϕRS603 genome showed strong similarity with those of Ralstonia phages ϕRSM1 and ϕRSM3, as reported by Askora et al. The ϕRS603 genome had no ORFs corresponding to ORFs 2, 3, 13 and 14 (integrase) of ϕRSM3. ϕRS603 had an ORF that was homologous to other Ralstonia phages ϕRSS0 and ϕRSS1; however, ϕRSM1 and ϕRSM3 did not.
Development of an Activity Card Sort for Australian adults aged 18–64 years
Authors: Gustafsson Louise , de Jonge Desleigh , Lai Yvonne , Muuse Jessica , Naude Nicola , Hoyle Melanie
Published: October 2014

Background/aim

The Activity Card Sort (ACS) is an interview‐based, client‐centred tool that measures the activity engagement of older adults aged 65 years and above. The tool consists of photos which clients are asked to sort into categories to reflect their previous and current level of engagement. The activities that older adults engage in may differ to that of younger adults and there is a need to explore the development of an ACS Australia for adults aged 18–64 years.

Methods

The Delphi survey method as applied by previous ACS development was used in this study. Purposeful and snowball sampling were employed to recruit participants between the ages of 18 and 64 years who were Australian citizens or residents. Demographic information was obtained via a questionnaire. There were two stages in the study. The first Delphi survey generated, selected and finalised activities and the second Delphi survey generated, selected and finalised categories.

Results

Australian citizens and residents were involved in the development of the activities (n = 61) and categories (n = 65). The final tool includes 85 activities sorted into three broad categories representing personal care, daily life and home maintenance; rest and relaxation; and high and low impact activities.

Conclusions

This study has resulted in the development of a preliminary version of an ACS for adults aged 18–64 years. Further work is required to determine the psychometric properties of the tool.
Reduced primary care respiratory infection visits following pregnancy and infancy vitamin D supplementation: a randomised controlled trial
Authors: Grant Cameron C. , Kaur Suhina , Waymouth Ellen , Mitchell Edwin A. , Scragg Robert , Ekeroma Alec , Stewart Alistair , Crane Julian , Trenholme Adrian , Camargo Carlos A.
Published: October 2014

Aim

To determine whether vitamin D supplementation reduces primary care visits for acute respiratory infection (ARI).

Methods

A randomised, double‐blind, placebo‐controlled trial was conducted in New Zealand and powered to determine the vitamin D dose needed to achieve normal vitamin D status during infancy. Healthy pregnant women, from 27 weeks’ gestation to birth, and their infants, from birth to age 6 months, were assigned to placebo or one of the two dosages of daily oral vitamin D3. Woman/infant pairs were randomised to placebo/placebo, 1000 IU/400 IU or 2000 IU/800 IU. For this ad hoc analysis, the primary care records of enrolled children were audited to age 18 months.

Results

Two hundred and sixty pregnant women were randomised to placebo (n = 87), lower‐dose (n = 87) or higher‐dose (n = 86) vitamin D3. In comparison with the placebo group (99%), the proportion of children making any ARI visits was smaller in the higher‐dose (87%, p = 0.004), but not the lower‐dose vitamin D3 group (95%, p = 0.17). The median number of ARI visits/child was less in the higher‐dose vitamin D3 group from age 6–18 months (placebo 4, lower dose 3, higher dose 2.5; p = 0.048 for higher‐dose vitamin D3 vs. placebo).

Conclusion

Vitamin D3 supplementation during pregnancy and infancy reduces primary care visits for ARI during early childhood.
Physiological weight loss chart helps to identify high‐risk infants who need breastfeeding support
Authors: Bertini Giovanna , Breschi Rita , Dani Carlo
Published: October 2014

Aim

Healthy, full‐term, exclusively breastfed infants are expected to lose weight in the first days after birth, but experts disagree about what constitutes a physiological neonatal weight loss and there is a lack of evidence‐based data. Our study aimed to construct a centile chart of neonatal weight loss in a healthy population of exclusively breastfed term neonates.

Methods

We retrospectively studied all infants born at an Italian centre that focused on natural childbirth from April 2007 to December 2012 and who complied with World Health Organization guidance on infant feeding. The infants’ weight loss was recorded after 12, 24, 36, 48, 60 and 72 h of life.

Results

We included 1760 healthy, full‐term, singleton babies born by vaginal delivery. Their mean weight loss was 5.95 ± 1.73%, 72.2% had maximal weight loss before discharge, only 3.9% lost more than 9% of their birthweight, and no infant lost more than 10%. We measured the percentage weight change from birth and each time the infant was examined, summarising how their weight varied in the first 72 h of life.

Conclusion

Our normative chart of physiological weight loss provided an important instrument for identifying high‐risk infants who required breastfeeding support.
Development of the germinal epithelium and early folliculogenesis during ovarian morphogenesis in the cichlid fish Cichlasoma dimerus (Teleostei, Perciformes)
Authors: Meijide Fernando J. , Rey Vázquez Graciela , Grier Harry J. , Lo Nostro Fabiana L. , Guerrero Graciela A.
Published: October 2014

Formation of the germinal epithelium and folliculogenesis during ovarian development in Cichlasoma dimerus were described at the light‐ and electron‐microscopic levels. Prior to gonadal differentiation, germ cells and enveloping support cells reside within an inpocketing of the coelomic epithelium. Separation of the germinal and interstitial compartments of the gonad by a basement membrane is apparent from early gonadal development. Upon ovarian differentiation, oogonia undergo cyst‐forming divisions leading to the formation of clusters of interconnected cystocytes that synchronously enter meiosis, becoming oocytes. At the pachytene step, each oocyte becomes individualized by cytoplasmic extensions of prefollicle cells, thereby developing as an ovarian follicle. Subsequent somatic reorganization leads to the formation of the ovarian lumen in a cephalo‐caudal gradient. As a result, the germinal epithelium becomes internalized and lines the ovarian lumen. As defined by its origin from the germinal epithelium, the ovarian follicle is composed of an oocyte and the surrounding follicle cells. Thecal cells derived from the stroma encompass the basement membrane outside the follicle, thus forming a follicle complex. A common basement membrane is shared by the germinal epithelium and the follicle complex along a small portion of its surface. This point of attachment represents the site at which the oocyte would be released to the ovarian lumen during ovulation.
Novel antibodies directed against the human erythropoietin receptor: creating a basis for clinical implementation
Authors: Maxwell Perry , Melendez‐Rodríguez Florinda , Matchett Kyle B. , Aragones Julian , Ben‐Califa Nathalie , Jaekel Heidelinde , Hengst Ludger , Lindner Herbert , Bernardini Andre , Brockmeier Ulf , Fandrey Joachim , Grunert Fritz , Oster Howard S. , Mittelman Moshe , El‐Tanani Mohamed , Thiersch Markus , Schneider Gasser Edith M. , Gassmann Max , Dangoor David , Cuthbert Robert J. , Irvine Alexandra , Jordan Anne , Lappin Terence , Thompson John , Neumann Drorit
Published: October 2014

Recombinant human erythropoietin (rHuEPO) is an effective treatment for anaemia but concerns that it causes disease progression in cancer patients by activation of EPO receptors (EPOR) in tumour tissue have been controversial and have restricted its clinical use. Initial clinical studies were flawed because they used polyclonal antibodies, later shown to lack specificity for EPOR. Moreover, multiple isoforms of EPOR caused by differential splicing have been reported in cancer cell lines at the mRNA level but investigations of these variants and their potential impact on tumour progression, have been hampered by lack of suitable antibodies. The EpoCan consortium seeks to promote improved pathological testing of EPOR, leading to safer clinical use of rHuEPO, by producing well characterized EPOR antibodies. Using novel genetic and traditional peptide immunization protocols, we have produced mouse and rat monoclonal antibodies, and show that several of these specifically recognize EPOR by Western blot, immunoprecipitation, immunofluorescence, flow cytometry and immunohistochemistry in cell lines and clinical material. Widespread availability of these antibodies should enable the research community to gain a better understanding of the role of EPOR in cancer, and eventually to distinguish patients who can be treated safely by rHuEPO from those at increased risk from treatment.
Wrong blood in tube – potential for serious outcomes: can it be prevented?
Authors: Bolton‐Maggs Paula H. B. , Wood Erica M. , Wiersum‐Osselton Johanna C.
Published: October 2014

‘Wrong blood in tube’ (WBIT) errors, where the blood in the tube is not that of the patient identified on the label, may lead to catastrophic outcomes, such as death from ABO‐incompatible red cell transfusion. Transfusion is a multistep, multidisciplinary process in which the human error rate has remained unchanged despite multiple interventions (education, training, competency testing and guidelines). The most effective interventions are probably the introduction of end‐to‐end electronic systems and a group‐check sample for patients about to receive their first transfusion, but neither of these eradicates all errors. Further longer term studies are required with assessment before and after introduction of the intervention. Although most focus has been on WBIT in relation to blood transfusion, all pathology samples should be identified and linked to the correct patient with the same degree of care. Human factors education and training could help to increase awareness of human vulnerability to error, particularly in the medical setting where there are many risk factors.
Incidence and risk factors for cataract after haematopoietic stem cell transplantation for childhood leukaemia: an LEA study
Authors: Horwitz Meryl , Auquier Pascal , Barlogis Vincent , Contet Audrey , Poiree Maryline , Kanold Justyna , Bertrand Yves , Plantaz Dominique , Galambrun Claire , Berbis Julie , Villes Virginie , Chastagner Pascal , Sirvent Nicolas , Oudin Claire , Michel Gérard
Published: October 2014

Cataract was prospectively assessed by serial slip lamp tests in 271 patients included in the Leucémie Enfants Adolescents (LEA) programme, the French cohort of childhood leukaemia survivors. All had received haematopoietic stem cell transplantation (HSCT) after total body irradiation (TBI, n = 201) or busulfan‐based (n = 70) myeloablative conditioning regimen. TBI was fractionated in all but six patients. The mean duration of follow‐up from HSCT was 10·3 years. Cataract was observed in 113/271 patients (41·7%); 9/113 (8·1%) needed surgery. Cumulative incidence after TBI increased over time from 30% at 5 years to 70·8% and 78% at 15 and 20 years, respectively, without any plateau thereafter. The 15‐year cumulative incidence was 12·5% in the Busulfan group. A higher cumulative steroid dose appeared to be a cofactor of TBI for cataract risk, in both univariate and multivariate Cox analysis. In the multivariate analysis, cataract had an impact in two quality of life domains: ‘the role limitation due to physical problems’ and ‘the role limitation due to emotional problems’. These data suggest that with increasing follow‐up, nearly all patients who receive TBI, even when fractionated, will suffer from cataract that can impact on their quality of life and that high cumulative steroid dose is a cofactor.
‘My great hope in life is to have a house, a family and a daughter’: relationships and sexuality in intellectually disabled people
Authors: Rojas Susana , Haya Ignacio , Lázaro‐Visa Susana
Published: October 2014

Accessible summary

Relationships and sexuality are important for disabled people.

Through the voices of 16 intellectually disabled adults, this paper shows what they consider to be important in areas such as education or personal relationships.

Most of the people interviewed would like to have a partner and live together as a couple.

Intellectually disabled people encounter many barriers to obtain certain basic rights.

This study starts from the premise that we are sexual beings, and therefore, sexuality is part of our lives and defines us as human beings. This is also true with regard to intellectually disabled people. Within the framework of broader qualitative research carried out in Spain, some partial results of an ongoing study aimed at finding out what a group of adults see as important in different areas of their lives are presented here. Through the personal narratives of 16 intellectually disabled people, this paper explores some of their ideas and demands about sexuality and their sexual lives. Most of the people interviewed expressed their desire to have a partner and to live as a couple. There is no doubt that many intellectually disabled people have greater control over their lives and the decisions that affect them, but as they tell us, the presence of old ideas can hinder them from securing their fundamental rights.
Repurposing Human PDE4 Inhibitors for Neglected Tropical Diseases. Evaluation of Analogs of the Human PDE4 Inhibitor GSK‐256066 as Inhibitors of PDEB1 of Trypanosoma brucei
Authors: Ochiana Stefan O. , Bland Nicholas D. , Settimo Luca , Campbell Robert K. , Pollastri Michael P.
Published: October 2014

Cyclic nucleotide phosphodiesterases (PDEs) have been identified as important enzyme targets for drug development in both humans and Trypanosoma brucei, the causative agent of human African trypanosomiasis. With this in mind, we recently reported the profiling of a range of human phosphodiesterase inhibitors, showing that human PDE4 inhibitors tend to display the best potency against the trypanosomal phosphodiesterase TbrPDEB1. Among these was GSK‐256066, a potent inhibitor of human PDE4 and a weak inhibitor of TbrPDEB1. In this report, we describe the results of a structure–activity relationship study of this chemotype, leading to the discovery of analogs with improved potency against TbrPDEB1 and micromolar inhibition of T. brucei cellular growth. We rationalize the potency trends via molecular docking of the new inhibitors into a recently reported apo structure of TbrPDEB1. The studies in this article will inform future efforts in repurposing human PDE inhibitors as antitrypanosomal agents.
Impact of human maxillary sinus volume on grafts dimensional changes used in maxillary sinus augmentation: a multislice tomographic study
Authors: Favato Mário N. , Vidigal Bruno C.L. , Cosso Maurício G. , Manzi Flávio R. , Shibli Jamil A. , Zenóbio Elton G.
Published: October 2014

Purpose

To assess the influence of complete maxillary sinus volume on the dimensional changes of different grafts used in maxillary sinus lift.

Materials and Methods

Analysis of 50 surgical procedures of maxillary sinus lift performed on 43 subjects using different grafting materials: fresh frozen allogenic particulated bone (11), hydroxyapatite (Endobon®) (17), 60% hydroxyapatite + 40% beta‐tricalcium phosphate (Bone Ceramic®) (12) and Bone Ceramic® + Emdogain® (10). One hundred and fifty multislice tomographic images of the maxillary sinus were obtained using the software Syngo CT 2011 A VOLUME, measuring complete maxillary sinus volume (T0) and dimensional changes of different graft materials during periods of 15 days (T1) and 180 days (T2). The factor studied was the influence of maxillary sinus volume on the dimensional changes of different graft materials used in maxillary sinus lift in patients with posterior edentulism. Data obtained were assessed using the Student's t‐test and Pearson's correlation coefficient.

Results

No correlation (r 0.112) between the total maxillary sinus volume and the dimensional changes of the different graft materials used in this study was observed (P > 0.05).

Conclusion

This study demonstrated that there is no sufficient evidence to support the thesis that the volume of the maxillary sinus influences the contraction of the grafts, at least on sample or the biomaterials evaluated in this cohort study.
Hb Wilde and Hb Patagonia: two novel elongated beta‐globin variants causing dominant beta‐thalassemia
Authors: Scheps Karen G. , Hasenahuer Marcia A. , Parisi Gustavo , Fornasari María S. , Pennesi Sandra P. , Erramouspe Beatriz , Basack Felisa N. , Veber Ernesto S. , Aversa Luis , Elena Graciela , Varela Viviana
Published: October 2014

We describe here the molecular and hematological characteristics of novel frameshift mutations in exon 2 of the HBB gene (in heterozygous state) found in two Argentinean pediatric patients with dominant β‐thalassemia‐like features. In Hb Wilde, HBB:c.270_273delTGAG(p.Glu90Cysfs*67), we detected the deletion of the third base of the codon 89 (T) and the codon 90 (GAG), whereas in Hb Patagonia, HBB:c.296_297dupGT(p.Asp99Trpfs*59), the frameshift mutation was due to a duplication of a ‘GT’ dinucleotide after the second base of codon 98 (GTG). The Hb Patagonia and Hb Wilde mutations would result in elongated β‐globin chains with modified C‐terminal sequences and a total of 155 and 157 amino acids residues, respectively. Based on bioinformatics and structural analysis, as well as protein modeling, we predict that the elongated β‐globins would affect the formation of the αβ dimers and their stability, which would further support the mechanism for the observed clinical features in both patients.
High level of full‐length cereblon mRNA in lower risk myelodysplastic syndrome with isolated 5q deletion is implicated in the efficacy of lenalidomide
Authors: Jonasova Anna , Bokorova Radka , Polak Jaroslav , Vostry Martin , Kostecka Arnost , Hajkova Hana , Neuwirtova Radana , Siskova Magda , Sponerova Dana , Cermak Jaroslav , Mikulenkova Dana , Cervinek Libor , Brezinova Jana , Michalova Kyra , Fuchs Ota
Published: October 2014

Downregulation of cereblon (CRBN) gene expression is associated with resistance to the immunomodulatory drug lenalidomide and poor survival outcomes in multiple myeloma (MM) patients. However, the importance of CRBN gene expression in patients with myelodysplastic syndrome (MDS) and its impact on lenalidomide therapy are not clear. In this study, we evaluate cereblon expression in mononuclear cells isolated from bone marrow [23 lower risk MDS patients with isolated 5q deletion (5q‐), 37 lower risk MDS patients with chromosome 5 without the deletion of long arms (non‐5q‐), and 24 healthy controls] and from peripheral blood (38 patients with 5q‐, 52 non‐5q‐ patients and 25 healthy controls) to gain insight into, firstly, the role of cereblon in lower risk MDS patients with or without 5q deletion and, secondly, into the mechanisms of lenalidomide action. Patients with 5q‐ lower risk MDS have the highest levels of CRBN mRNA in comparison with both lower risk MDS without the deletion of long arms of chromosome 5 and healthy controls. CRBN gene expression was measured using the quantitative TaqMan real‐time PCR. High levels of CRBN mRNA were detected in all lenalidomide responders during the course of therapy. A significant decrease of the CRBN mRNA level during lenalidomide treatment is associated with loss of response to treatment and disease progression. These results suggest that, similar to the treatment of MM, high levels of full‐length CRBN mRNA in lower risk 5q‐ patients are necessary for the efficacy of lenalidomide.
Reduced sensitivity to both positive and negative reinforcement in mice over‐expressing the 5‐hydroxytryptamine transporter
Authors: Line Samantha J. , Barkus Chris , Rawlings Nancy , Jennings Katie , McHugh Stephen , Sharp Trevor , Bannerman David M.
Published: October 2014

The 5‐hydroxytryptamine (5‐HT) transporter (5‐HTT) is believed to play a key role in both normal and pathological psychological states. Much previous data suggest that the s allele of the polymorphic regulatory region of the 5‐HTT gene promoter is associated with reduced 5‐HTT expression and vulnerability to psychiatric disorders, including anxiety and depression. In comparison, the l allele, which increases 5‐HTT expression, is generally considered protective. However, recent data link this allele to both abnormal 5‐HT signalling and psychopathic traits. Here, we studied the processing of aversive and rewarding cues in transgenic mice that over‐express the 5‐HTT (5‐HTTOE mice). Compared with wild‐type mice, 5‐HTTOE mice froze less in response to both a tone that had previously been paired with footshock, and the conditioning context. In addition, on a decision‐making T‐maze task, 5‐HTTOE mice displayed reduced preference for a larger, delayed reward and increased preference for a smaller, immediate reward, suggesting increased impulsiveness compared with wild‐type mice. However, further inspection of the data revealed that 5‐HTTOE mice displayed a relative insensitivity to reward magnitude, irrespective of delay. In contrast, 5‐HTTOE mice appeared normal on tests of spatial working and reference memory, which required an absolute choice between options associated with either reward or no reward. Overall, the present findings suggest that 5‐HTT over‐expression results in a reduced sensitivity to both positive and negative reinforcers. Thus, these data show that increased 5‐HTT expression has some maladaptive effects, supporting recent suggestions that l allele homozygosity may be a potential risk factor for disabling psychiatric traits.
Amide hydrogen exchange in HIV–1 subtype B and C proteases – insights into reduced drug susceptibility and dimer stability
Authors: Naicker Previn , Stoychev Stoyan , Dirr Heini W. , Sayed Yasien
Published: October 2014

Since its identification, HIV has continued to have a detrimental impact on the lives of millions of people throughout the world. The protease of HIV is a major target in antiviral treatment. The South African HIV–1 subtype C (C–SA) protease displays weaker binding affinity for some clinically approved protease inhibitors in comparison with the HIV–1 subtype B protease. The heavy HIV burden in sub‐Saharan Africa, where subtype C HIV–1 predominates, makes this disparity a topic of great interest. In light of this, the enzyme activity and affinity of protease inhibitors for the subtype B and C–SA proteases were determined. The relative vitality, indicating the selective advantage of polymorphisms, of the C–SA protease relative to the subtype B protease in the presence of ritonavir and darunavir was four‐ and tenfold greater, respectively. Dynamic differences that contribute to the reduced drug susceptibility of the C–SA protease were investigated by performing hydrogen–deuterium exchange/mass spectrometry (HDX/MS) on unbound subtype B and C–SA proteases. The reduced propensity to form the E35–R57 salt bridge, and alterations in the hydrophobic core of the C–SA protease, are proposed to affect the anchoring of the flexible flaps, resulting in an increased proportion of the fully open flap conformation. HDX/MS data suggested that the N–terminus of both proteases is less stable than the C–terminus of the proteases, thus explaining the increased efficacy of dimerization inhibitors targeted toward the C–terminus of HIV proteases. As far as we are aware, this is the first report on assessment of HIV protease dynamics using HDX/MS.
NLS copy‐number variation governs efficiency of nuclear import – case study on dUTPases
Authors: Róna Gergely , Pálinkás Hajnalka L. , Borsos Máté , Horváth András , Scheer Ildikó , Benedek András , Nagy Gergely N. , Zagyva Imre , Vértessy Beáta G.
Published: October 2014

Nucleocytoplasmic trafficking of large macromolecules requires an active transport machinery. In many cases, this is initiated by binding of the nuclear localization signal (NLS) peptide of cargo proteins to importin‐α molecules. Fine orchestration of nucleocytoplasmic trafficking is of particularly high importance for proteins involved in maintenance of genome integrity, such as dUTPases, which are responsible for prevention of uracil incorporation into the genome. In most eukaryotes, dUTPases have two homotrimeric isoforms: one of these contains three NLSs and is present in the cell nucleus, while the other is located in the cytoplasm or the mitochondria. Here we focus on the unusual occurrence of a pseudo‐heterotrimeric dUTPase in Drosophila virilis that contains one NLS, and investigate its localization pattern compared to the homotrimeric dUTPase isoforms of Drosophila melanogaster. Although the interaction of individual NLSs with importin‐α has been well characterized, the question of how multiple NLSs of oligomeric cargo proteins affect their trafficking has been less frequently addressed in adequate detail. Using the D. virilis dUTPase as a fully relevant physiologically occurring model protein, we show that NLS copy number influences the efficiency of nuclear import in both insect and mammalian cell lines, as well as in D. melanogaster and D. virilis tissues. Biophysical data indicate that NLS copy number determines the stoichiometry of complexation between importin‐α and dUTPases. The main conclusion of our study is that, in D. virilis, a single dUTPase isoform efficiently reproduces the cellular dUTPase distribution pattern that requires two isoforms in D. melanogaster.

Structured digital abstract

• ABC dUTPase and importin-a bind by molecular sieving (View interaction) • AAA dUTPase and importin-a bind by molecular sieving (View interaction) • ABC dUTPase and importin-a bind by comigration in non denaturing gel electrophoresis (View interaction) • AAA dUTPase and importin-a bind by comigration in non denaturing gel electrophoresis (View interaction) • ABC dUTPase and importin-a bind by isothermal titration calorimetry (View interaction)

• AAA dUTPase and importin-a  bind by isothermal titration calorimetry (View interaction)
Effects of adding acetic acid and molasses on fermentation quality and aerobic stability of total mixed ration silage prepared with hulless barley straw in Tibet
Authors: Qiu Xiaoyan , Guo Gang , Yuan Xianjun , Shao Tao
Published: October 2014

In order to enlarge the feed resources in Tibet, hulless barley (Hordeum vulgare L.) straw was conserved as total mixed ration (TMR) silage by combining it with whole crop oat (Avena sativa L.), alfalfa (Medicago sativa L.) and concentrate. This experiment was carried out to study the effects of acetic acid or/and molasses on the quality of TMR silages prepared with hulless barley straw. Treatments included: control (C) silage without additive, or silage with acetic acid (A, 0.3% fresh matter basis), molasses (M, 3% fresh matter basis) and A+M. Triplicate silos were opened after 45 days of ensiling and chemical and microbial analyses were conducted on the samples. Aerobic stability was tested 2, 4 and 6 days after exposure to air. Silages of all four TMR treatments were of good quality with high lactic acid (LA) concentration and low ratios of ammonia nitrogen in total nitrogen and butyric acid contents. Both the M and A+M silages had higher concentrations of LA (P < 0.05) and lower pH (P < 0.05) than the C and A silages. During aerobic exposure, aerobic deterioration occurred in the C and M silages with air infiltration. Resistance to deterioration was sustained in the A and A+M silages, with high concentration of acetic acid and low population (~4 log cfu g−1) of yeast throughout the 6‐day test. Furthermore, the A+M treatment showed a slow change in pH and LA in comparison to the A treatment. These findings indicate that adding acetic acid inhibited the yeast growth and improved the aerobic stability. The present study suggested that A+M addition was effective for improving both the fermentation quality and the aerobic stability of TMR silages containing hulless barley straw.
Determination of Retained B2O3 Content in ZrB2‐30 vol% SiC Oxide Scales
Authors: Shugart Kathleen , Liu Siying , Craven Forrest , Opila Elizabeth
Published: October 2014

The composition of the borosilicate glass layer formed during oxidation of ZrB2‐30 vol% SiC was determined to elucidate the extent of B2O3 retention in the oxide during high‐temperature oxidation. Oxidation was conducted in stagnant air at 1300°C, 1400°C, and 1500°C for times between 100 and 221 min. Specimens were characterized using mass change and scanning electron microscopy. After oxidation, the borosilicate glass layer was dissolved from the specimens sequentially with deionized H2O and HF acid, to analyze the glass composition using inductively coupled plasma optical emission spectrometry. It was found that the average B2O3 content in the glass scale ranged from 23 to 47 mol%. Retained B2O3 content in the bulk of the glass decreased with increasing temperature, confirming increased volatility with temperature. Boron depth profiles were also obtained in the near surface region using X‐ray photoelectron spectroscopy and energy dispersive spectroscopy. The measured B concentrations were used to estimate the B2O3 concentration profile and B diffusion coefficients in the borosilicate glass. Implications for the ZrB2‐SiC oxidation process are discussed.
Solid‐State CMAS Corrosion of an EB‐PVD YSZ Coated Turbine Blade: Zr4+ Partitioning and Phase Evolution
Authors: Mechnich Peter , Braue Wolfgang
Published: October 2014

The corrosion of an ex‐service 7‐YSZ coated high‐pressure turbine blade by Si‐undersaturated, Fe‐,Ti‐rich CMAS (FT‐CMAS) was investigated. The constituents of the FT‐CMAS deposit and its synthetic laboratory counterpart are identified by means of analytical SEM and XRD. FT‐CMAS mainly consists of rhönite‐, melilite‐, and clinopyroxene‐type solid solutions. FT‐CMAS reacts with the 7‐YSZ thermal barrier coating to a continuous layer of a Ca,Zr‐rich garnet phase also known as kimzeyite. The microstructural analysis indicates that kimzeyite has formed via solid‐state reaction. The formation of kimzeyite was studied in laboratory using YSZ/FT‐CMAS diffusion couples and powder compacts. Experiments provide strong evidence that kimzeyite is forming predominantly from YSZ and melilite, whereas clinopyroxene and especially rhönite appear relative stable versus YSZ. The preferred kimzeyite formation from melilite and YSZ is explained by the availability of sixfold coordinated lattice sites for Zr4+ on melilite decomposition and subsequent garnet crystallization.
The Temperature‐Dependent Ideal Tensile Strength of ZrB2, HfB2, and TiB2
Authors: Cheng Tianbao , Li Weiguo
Published: October 2014

This paper is aimed to fill the void in the literature regarding the temperature‐dependent ideal tensile strength of the AlB2‐like diborides. To this end, two models of temperature‐dependent ideal tensile strength of hexagonal single crystals are developed based on the critical failure energy density principle and the critical strain principle. The two models relate the ideal tensile strength to the elastic properties, thermal expansion, and specific heat at constant pressure of single crystals. The temperature‐dependent uniaxial ideal tensile strengths of ZrB2, HfB2, and TiB2 are predicted. The predictions from the critical strain principle agree well with that from the critical failure energy density principle. The study shows that the temperature dependence of the ideal tensile strength is similar to that of Young's modulus. That is, the ideal tensile strength firstly remains approximately constant and then decreases linearly as temperature increases. The fracture failure for hexagonal single crystals at elevated temperatures is identified as a strain‐controlled criterion for the first time.
Surface Chemistry of Calcium Aluminosilicate Glasses
Authors: Snellings Ruben
Published: October 2014

The surface chemistry of synthetic calcium aluminosilicate glasses exposed to aqueous solutions of varying pH was described using zeta potential measurements and batch surface titrations. Element release and proton consumption were measured to characterize the reactions at the surface as a function of pH. It was found that proton–metal exchange or leaching was the dominant proton consumption process at low pH. The exchange reaction was observed to maintain charge balance, indicating that over the limited duration of the experiments no repolymerization of the silica‐rich leached layer occurred. At high pH, dissolution and hydrolysis of aqueous ions controlled the proton mass balance, no evidence for the formation of leached layers was found. Silica‐rich glasses were found to be more resistant to corrosion by either proton–metal exchange or dissolution than CaO‐rich glasses. Glass basicity was found to be reflected in the establishment of a higher pH when immersing the glass in neutral aqueous solutions and in a reduced stability of the glass surface toward aqueous solutions. High pH, high surface charge and reduced glass network polymerization act together in enhancing the reaction of CaO‐rich glasses with aqueous solutions.
Volume Expansion Caused by Water Penetration into Silica Glass
Authors: Wiederhorn Sheldon M. , Yi Feng , LaVan David , Richter Lee J. , Fett Theo , Hoffmann Michael J.
Published: October 2014

By measuring the curvature of thin disks of vitreous silica that have been penetrated by water from one side only, we determined the volume expansion of the silica and the effect of this volume expansion on its strength. We found that the water‐strengthening process depended on crack‐size, temperature, and the amount of swelling of the silica. We also evaluated the diffusivity of water in vitreous silica, using the swelling stresses as the diffusion metric. Diffusivity values, so obtained, are close to the accepted values for the diffusion of water in vitreous silica, as is the activation energy for the diffusion process. Our data suggest that swelling and the consequent bending of the disks is caused by silanol group formation in the silica structure; molecular water plays little role in the swelling process.