bluuebunny/crossref_metadata_2025 huggingface dataset:
HIV-1 infection of hematopoietic progenitor cells in vivo in humanized mice
Authors: Nixon Christopher C. , Vatakis Dimitrios N. , Reichelderfer Scott N. , Dixit Dhaval , Kim Sohn G. , Uittenbogaart Christel H. , Zack Jerome A.
Published: July 2013

Key Points

Some CD34+CD38+ intermediate hematopoietic progenitor cells express HIV-1 entry receptors and are susceptible to direct infection by HIV. Blood progenitors from HIV-exposed, humanized BLT mice show impaired hematopoietic potential and give rise to progeny that harbor provirus.
HTR4 gene structure and altered expression in the developing lung
Authors: Hodge Emily , Nelson Carl P , Miller Suzanne , Billington Charlotte K , Stewart Ceri E , Swan Caroline , Malarstig Anders , Henry Amanda P , Gowland Catherine , Melén Erik , Hall Ian P , Sayers Ian
Published: July 2013

Background

Meta-analyses of genome-wide association studies (GWAS) have identified single nucleotide polymorphisms (SNPs) spanning the 5-hydroxytryptamine receptor 4 (5-HT4R) gene (HTR4) associated with lung function. The aims of this study were to i) investigate the expression profile of HTR4 in adult and fetal lung tissue and cultured airway cells, ii) further define HTR4 gene structure and iii) explore the potential functional implications of key SNPs using a bioinformatic approach.

Methods

Following reverse transcription (RT)-PCR in human brain, 5′ rapid amplification of cDNA ends (5′ RACE) was used to examine the exonic structure of HTR4 at the 5′ end. Quantitative (Q)-PCR was used to quantify HTR4 mRNA expression in total RNA from cultured airway cells and whole lung tissue. Publically available gene microarray data on fetal samples of estimated gestational age 7–22 weeks were mined for HTR4 expression. Immunohistochemistry (IHC; in adult and fetal lung tissue) and a radioligand binding assay (in cultured airway cells) were used to analyze 5­HT4R protein expression.

Results

IHC in adult lung, irrespective of the presence of chronic obstructive pulmonary disease (COPD), suggested low level expression of 5-HT4R protein, which was most prominent in alveolar pneumocytes. There was evidence of differential 5-HT4R protein levels during gestation in fetal lung, which was also evident in gene expression microarray data. HTR4 mRNA expression, assessed by Q-PCR, was <0.5% relative to brain in total adult lung tissue and in human airway smooth muscle (HASM) and bronchial epithelial cells (HBEC) derived from adult donors. Radioligand binding experiments also indicated that HBEC and HASM cells did not express a significant 5-HT4R population. 5′ RACE in brain identified a novel N-terminal variant, containing an extended N-terminal sequence. The functional significance of key HTR4 SNPs was investigated using the encyclopedia of DNA elements consortium (ENCODE) dataset. These analyses identified multiple alterations in regulatory motifs for transcription factors implicated in lung development, including Foxp1.

Conclusions

Taken together, these data suggest a role for HTR4 in lung development, which may at least in part explain the genetic association with lung function.
KEGG orthology-based annotation of the predicted proteome of Acropora digitifera: ZoophyteBase - an open access and searchable database of a coral genome
Authors: Dunlap Walter C , Starcevic Antonio , Baranasic Damir , Diminic Janko , Zucko Jurica , Gacesa Ranko , H van Oppen Madeleine J , Hranueli Daslav , Cullum John , Long Paul F
Published: July 2013

Background

Contemporary coral reef research has firmly established that a genomic approach is urgently needed to better understand the effects of anthropogenic environmental stress and global climate change on coral holobiont interactions. Here we present KEGG orthology-based annotation of the complete genome sequence of the scleractinian coralAcropora digitiferaand provide the first comprehensive view of the genome of a reef-building coral by applying advanced bioinformatics.

Description

Sequences from the KEGG database of protein function were used to construct hidden Markov models. These models were used to search the predicted proteome ofA. digitiferato establish complete genomic annotation. The annotated dataset is published in ZoophyteBase, an open access format with different options for searching the data. A particularly useful feature is the ability to use a Google-like search engine that links query words to protein attributes. We present features of the annotation that underpin the molecular structure of key processes of coral physiology that include (1) regulatory proteins of symbiosis, (2) planula and early developmental proteins, (3) neural messengers, receptors and sensory proteins, (4) calcification and Ca2+-signalling proteins, (5) plant-derived proteins, (6) proteins of nitrogen metabolism, (7) DNA repair proteins, (8) stress response proteins, (9) antioxidant and redox-protective proteins, (10) proteins of cellular apoptosis, (11) microbial symbioses and pathogenicity proteins, (12) proteins of viral pathogenicity, (13) toxins and venom, (14) proteins of the chemical defensome and (15) coral epigenetics.

Conclusions

We advocate that providing annotation in an open-access searchable database available to the public domain will give an unprecedented foundation to interrogate the fundamental molecular structure and interactions of coral symbiosis and allow critical questions to be addressed at the genomic level based on combined aspects of evolutionary, developmental, metabolic, and environmental perspectives.
Susceptibility (risk and protective) factors for in-patient violence and self-harm: prospective study of structured professional judgement instruments START and SAPROF, DUNDRUM-3 and DUNDRUM-4 in forensic mental health services
Authors: Abidin Zareena , Davoren Mary , Naughton Leena , Gibbons Olivia , Nulty Andrea , Kennedy Harry G
Published: July 2013

Background

The START and SAPROF are newly developed fourth generation structured professional judgement instruments assessing strengths and protective factors. The DUNDRUM-3 and DUNDRUM-4 also measure positive factors, programme completion and recovery in forensic settings.

Methods

We compared these instruments with other validated risk instruments (HCR-20, S-RAMM), a measure of psychopathology (PANSS) and global function (GAF). We prospectively tested whether any of these instruments predict violence or self harm in a secure hospital setting (n = 98) and whether they had true protective effects, interacting with and off-setting risk measures.

Results

SAPROF and START-strengths had strong inverse (negative) correlations with the HCR-20 and S-RAMM. SAPROF correlated strongly with GAF (r = 0.745). In the prospective in-patient study, SAPROF predicted absence of violence, AUC = 0.847 and absence of self-harm AUC = 0.766. START-strengths predicted absence of violence AUC = 0.776, but did not predict absence of self-harm AUC = 0.644. The DUNDRUM-3 programme completion and DUNDRUM-4 recovery scales also predicted in-patient violence (AUC 0.832 and 0.728 respectively), and both predicted in-patient self-harm (AUC 0.750 and 0.713 respectively). When adjusted for the HCR-20 total score however, SAPROF, START-S, DUNDRUM-3 and DUNDRUM-4 scores were not significantly different for those who were violent or for those who self harmed. The SAPROF had a significant interactive effect with the HCR-dynamic score. Item to outcome studies often showed a range of strengths of association with outcomes, which may be specific to the in-patient setting and patient group studied.

Conclusions

The START and SAPROF, DUNDRUM-3 and DUNDRUM-4 can be used to assess both reduced and increased risk of violence and self-harm in mentally ill in-patients in a secure setting. They were not consistently better than the GAF, HCR-20, S-RAMM, or PANSS when predicting adverse events. Only the SAPROF had an interactive effect with the HCR-20 risk assessment indicating a true protective effect but as structured professional judgement instruments all have additional content (items) complementary to existing risk assessments, useful for planning treatment and risk management.
Biomechanical comparison of anterior lumbar interbody fusion: stand-alone interbody cage versus interbody cage with pedicle screw fixation - a finite element analysis
Authors: Choi Kyung-Chul , Ryu Kyeong-Sik , Lee Sang-Ho , Kim Yeong Hyeon , Lee Sung Jae , Park Chun-Kun
Published: July 2013

Background

Anterior lumbar interbody fusion (ALIF) followed by pedicle screw fixation (PSF) is used to restore the height of the intervertebral disc and provide stability. Recently, stand-alone interbody cage with anterior fixation has been introduced, which eliminates the need for posterior surgery. We compared the biomechanics of the stand-alone interbody cage to that of the interbody cage with additional PSF in ALIF.

Methods

A three-dimensional, non-linear finite element model (FEM) of the L2-5 segment was modified to simulate ALIF in L3-4. The models were tested under the following conditions: (1) intact spine, (2) destabilized spine, (3) with the interbody cage alone (type 1), (4) with the stand-alone cage with anterior fixation (SynFix-LR®; type 2), and (5) with type 1 in addition to PSF (type 3). Range of motion (ROM) and the stiffness of the operated level, ROM of the adjacent segments, load sharing distribution, facet load, and vertebral body stress were quantified with external loading.

Results

The implanted models had decreased ROM and increased stiffness compared to those of the destabilized spine. The type 2 had differences in ROM limitation of 8%, 10%, 4%, and 6% in flexion, extension, axial rotation, and lateral bending, respectively, compared to those of type 3. Type 2 had decreased ROM of the upper and lower adjacent segments by 3-11% and 3-6%, respectively, compared to those of type 3. The greatest reduction in facet load at the operated level was observed in type 3 (71%), followed by type 2 (31%) and type 1 (23%). An increase in facet load at the adjacent level was highest in type 3, followed by type 2 and type 1. The distribution of load sharing in type 2 (anterior:posterior, 95:5) was similar to that of the intact spine (89:11), while type 3 migrated posterior (75:25) to the normal. Type 2 reduced about 15% of the stress on the lower vertebral endplate compared to that in type 1. The stress of type 2 increased two-fold compared to the stress of type 3, especially in extension.

Conclusions

The stand-alone interbody cage can provide sufficient stability, reduce stress in adjacent levels, and share the loading distribution in a manner similar to an intact spine.
Single nucleotide polymorphisms associated with non-contact soft tissue injuries in elite professional soccer players: influence on degree of injury and recovery time
Authors: Pruna Ricard , Artells Rosa , Ribas Jordi , Montoro Bruno , Cos Francesc , Muñoz Carme , Rodas Gil , Maffulli Nicola
Published: July 2013

Background

The biological mechanisms involved in non-contact musculoskeletal soft tissue injuries (NCMSTI) are poorly understood. Genetic risk factors may be associated with susceptibility to injuries, and may exert marked influence on recovery times.

Methods

Data on type and degree of injury and recovery time were collected in 73 male professional soccer players (43 White, 11 Black Africans and 19 Hispanics) who suffered total of 242 injuries (203 muscle, 24 ligament, and 15 tendon injuries). One single nucleotide polymorphism (SNPs) in the following genes were analyzed: Elastin (ELN); Titin (TTN); SRY-related HMG-box (SOX15); Insulin-like growth factor 2 (IGF2); Chemokine, CC motif, ligand 2 (CCL2); Collagen type 1 alpha 1(COL1A1); Collagen type 5 alpha 1 (COL5A1), and Tenascin C (TNC).

Results

There was evidence of a statistically significant association between the degree of injury and the IGF2 genotype (P = 0.034). In addition, there was evidence of a statistically significant association between the degree of muscle injury and CCL2 (P = 0.026) Finally, there was evidence of a statistically significant association between ELN and degree of injury (p = 0.009) and recovery time (P = 0.043). There was no evidence of a statistically significant association between any of the genes studied and degree of injury or recovery time for tendon injuries.

Conclusion

SNPs in the IGF2, CCL2, and ELN genes may be associated to the degree and recovery time of NCMSTI.
An efficient protocol for the purification and labeling of entire yeast septin rods from E.coli for quantitative in vitroexperimentation
Authors: Renz Christian , Johnsson Nils , Gronemeyer Thomas
Published: July 2013

Background

The detailed understanding of the functions and mechanisms of the actin and microtubuli cytoskeleton depended, besides innovative methods in live cell imaging, on the purification and labeling of its constituents. This allowed researchers to quantitatively measure filament stability, the rates of filament turnover as well as the determination of the influence of cofactors on filament formation and structure. Septins form the least understood class of cytoskeletal structures in nearly all eukaryotic cells so far examined. In yeast, they comprise a family of proteins (Cdc3, Cdc10, Cdc11, Cdc12, Shs1) that form a co-polymeric, ring-like structure beneath the membrane. This ring serves as a template for the formation of a new bud neck and as a landing pat for proteins involved in polar growth and cytokinesis. Further progress in investigating the mechanisms of septin-structure formation and regulation is hampered by the lack of protocols to modify homogenous samples of purified septins with useful probes for in vitro biochemical studies.

Results

We present a protocol for the purification and labeling of yeast septin rods. The four individual septin subunits were co-expressed in E.coli. One subunit of the septin polymer was expressed as SNAP tag fusion protein allowing for rapid and stoichiometric labeling with derivatized Benzylguanine (BG). To demonstrate the applicability of our approach, we introduced two different SNAP tag substrates: septin rods labeled with fluorescent BG compounds enabled us to monitor the formation of filaments by fluorescence microscopy whereas BG-biotin was used to couple septin rods to a sensor chip for quantitative surface plasmon resonance binding experiments. In a first application, we determined the affinity and the binding kinetics of the yeast protein Bni5 to the individually coupled septin rods. In a further application we could demonstrate that a once formed septin rod hardly exchange its subunits.

Conclusions

The herein introduced protocol of purifying SNAP tag modified septins from E.coli allowed us to derivatize the obtained septin rods with probes for the further in vitro characterization of this class of cytoskeletal elements. The availability of a very diverse set of SNAP tag substrates should open the way to investigate different aspects of septin biochemistry in mechanistic detail.
Effects of cocktail of four local Malaysian medicinal plants (Phyllanthus spp.) against dengue virus 2
Authors: Lee Sau Har , Tang Yin Quan , Rathkrishnan Anusyah , Wang Seok Mui , Ong Kien Chai , Manikam Rishya , Payne Bobby Joe , Jaganath Indu Bala , Sekaran Shamala Devi
Published: July 2013

Background

The absence of commercialized vaccines and antiviral agents against dengue has made the disease a major health concern around the world. With the current dengue virus transmission rate and incidences, the development of antiviral drugs is of vital need. The aim of this project was to evaluate the possibility of developing a local medicinal plant, Phyllanthus as an anti-dengue agent.

Methods

Cocktail (aqueous and methanolic) extracts were prepared from four species of Phyllanthus (P.amarus, P.niruri, P.urinaria, and P.watsonii) and their polyphenolic compounds were identified via HPLC and LC-MS/MS analysis. MTS assay was then carried out to determine the maximal non-toxic dose (MNTD) of the extracts, followed by screening of the in vitro antiviral activity of aqueous cocktail extracts against DENV2 by means of time-of-addition (pre-, simultaneous and post-) using RT-qPCR. The differentially expressed proteins in the treated and infected cells were analysed with two dimensional gel electrophoresis experiments.

Results

Several active compounds including gallic acid, geraniin, syringin, and corilagen have been identified. The MNTD of both aqueous and methanolic extracts on Vero cells were 250.0 μg/ml and 15.63 μg/ml respectively. Phyllanthus showed strongest inhibitory activity against DENV2 with more than 90% of virus reduction in simultaneous treatment. Two-dimensional analysis revealed significantly altered levels of thirteen proteins, which were successfully identified by tandem MS (MS/MS). These altered proteins were involved in several biological processes, including viral entry, viral transcription and translation regulations, cytoskeletal assembly, and cellular metabolisms.

Conclusions

Phyllanthus could be potentially developed as an anti-DENV agent.
Global informetric perspective studies on translational medical research
Authors: Yao Qiang , Lyu Peng-Hui , Ma Fei-Cheng , Yao Lan , Zhang Shi-Jing
Published: July 2013

Background

Translational medical research literature has increased rapidly in the last few decades and played a more and more important role during the development of medicine science. The main aim of this study is to evaluate the global performance of translational medical research during the past few decades.

Methods

Bibliometric, social network analysis, and visualization technologies were used for analyzing translational medical research performance from the aspects of subject categories, journals, countries, institutes, keywords, and MeSH terms. Meanwhile, the co-author, co-words and cluster analysis methods were also used to trace popular topics in translational medical research related work.

Results

Research output suggested a solid development in translational medical research, in terms of increasing scientific production and research collaboration. We identified the core journals, mainstream subject categories, leading countries, and institutions in translational medical research. There was an uneven distribution of publications at authorial, institutional, and national levels. The most commonly used keywords that appeared in the articles were “translational research”, “translational medicine”, “biomarkers”, “stroke”, “inflammation”, “cancer”, and “breast cancer”.

Conclusions

The subject categories of “Research & Experimental Medicine”, “Medical Laboratory Technology”, and “General & Internal Medicine” play a key role in translational medical research both in production and in its networks. Translational medical research and CTS, etc. are core journals of translational research. G7 countries are the leading nations for translational medical research. Some developing countries, such as P.R China, also play an important role in the communication of translational research. The USA and its institutions play a dominant role in the production, collaboration, citations and high quality articles. The research trends in translational medical research involve drug design and development, pathogenesis and treatment of disease, disease model research, evidence-based research, and stem and progenitor cells.
A characterization of four B16 murine melanoma cell sublines molecular fingerprint and proliferation behavior
Authors: Danciu Corina , Falamas Alexandra , Dehelean Cristina , Soica Codruta , Radeke Heinfried , Barbu-Tudoran Lucian , Bojin Florina , Pînzaru Simona Cîntă , Munteanu Melania F
Published: July 2013

Background

One of the most popular and versatile model of murine melanoma is by inoculating B16 cells in the syngeneic C57BL6J mouse strain. A characterization of different B16 modified cell sub-lines will be of real practical interest. For this aim, modern analytical tools like surface enhanced Raman spectroscopy/scattering (SERS) and MTT were employed to characterize both chemical composition and proliferation behavior of the selected cells.

Methods

High quality SERS signal was recorded from each of the four types of B16 cell sub-lines: B164A5, B16GMCSF, B16FLT3, B16F10, in order to observe the differences between a parent cell line (B164A5) and other derived B16 cell sub-lines. Cells were incubated with silver nanoparticles of 50–100 nm diameter and the nanoparticles uptake inside the cells cytoplasm was proved by transmission electron microscopy (TEM) investigations. In order to characterize proliferation, growth curves of the four B16 cell lines, using different cell numbers and FCS concentration were obtained employing the MTT proliferation assay. For correlations doubling time were calculated.

Results

SERS bands allowed the identification inside the cells of the main bio-molecular components such as: proteins, nucleic acids, and lipids. An "on and off" SERS effect was constantly present, which may be explained in terms of the employed laser power, as well as the possible different orientations of the adsorbed species in the cells in respect to the Ag nanoparticles. MTT results showed that among the four tested cell sub-lines B16 F10 is the most proliferative and B164A5 has the lower growth capacity. Regarding B16FLT3 cells and B16GMCSF cells, they present proliferation ability in between with slight slower potency for B16GMCSF cells.

Conclusion

Molecular fingerprint and proliferation behavior of four B16 melanoma cell sub-lines were elucidated by associating SERS investigations with MTT proliferation assay.
Intelligent spectrum assignment and migration in cognitive radio network
Authors: Liu Zhiyong Michael , Nasser Nidal , Hassanein Hossam S
Published: July 2013

Cognitive radio networks significantly improve spectrum utilization efficiency by allowing unlicensed users to opportunistically access the unused licensed spectrum. One of the key challenges is to determine how to properly assign the unused spectrum slots without impacting the licensed users and if necessary, move the existing users to different spectrum bands. In this paper, we present a spectrum migration model and propose a simulated annealing approach to determine the proper spectrum migration solution, which optimizes the aggregated utility while satisfying capacity constraints, interference constraints, and rate requirement constraints. The algorithm also takes into account different classes of traffic. We show that the algorithm greatly outperforms the greedy migration approach and other assignment approaches without migration.
The role of community-based health planning and services strategy in involving males in the provision of family planning services: a qualitative study in Southern Ghana
Authors: Adongo Philip Baba , Tapsoba Placide , Phillips James F , Tabong Philip Teg-Nefaah , Stone Alison , Kuffour Emmanuel , Esantsi Selina F , Akweongo Patricia
Published: July 2013

Background

Reproductive health and Family Planning (FP) services have been of global concern especially in developing countries where fertility rates are high. Traditionally FP services had always targeted females with little or no attention given to males. To ensure equitable distribution of health services, Ministry of Health (MOH), Ghana adopted the Community-Based Health Planning and Services (CHPS) as a nationwide health policy with the aim of reducing obstacles to physical and geographical access to health care delivery including FP services. However, not much is known about the extent to which this policy has contributed to male involvement in FP services. This qualitative descriptive study was therefore designed to explore male involvement in FP services in communities with well functioning CHPS and those with less or no functioning CHPS structures. The study further solicited views of the community on the health status of children.

Methods

This was a qualitative descriptive study and adapted the design of an ongoing study to assess the impact of male involvement in FP referred to as the Navrongo experiment in Northern Ghana. Twelve focus group discussions were held with both male and female community members, six in communities with functional CHPS and six for communities with less/no-functional CHPS. In addition, fifty- nine (59) in-depth interviews were held with Community Health Officers (CHOs), Community Health Volunteers (CHVs) and Health Managers at both the districts and regional levels. The interviews and discussions were tape recorded digitally, transcribed and entered into QSR Nvivo 10© for analysis.

Results

The results revealed a general high perception of an improved health status of children in the last ten years in the communities. These improvements were attributed to immunization of children, exclusive breastfeeding, health education given to mothers on childcare, growth monitoring of children and accessible health care. Despite these achievements in the health of children, participants reported that malnutrition was still rife in the community. The results also revealed that spousal approval was still relevant for women in the use of contraceptives; however, the matrilineal system appears to give more autonomy to women in decision-making. The CHPS strategy was perceived as very helpful with full community participation at all levels of the implementation process. Males were more involved in FP services in communities with functioning CHPS than those without functioning CHPS.

Conclusion

The CHPS strategy has increased access to FP services but spousal consent was very important in the use of FP services. Involving males in reproductive health issues including FP is important to attain reproductive health targets.
Linking melanism to brain development: expression of a melanism-related gene in barn owl feather follicles covaries with sleep ontogeny
Authors: Scriba Madeleine F , Ducrest Anne-Lyse , Henry Isabelle , Vyssotski Alexei L , Rattenborg Niels C , Roulin Alexandre
Published: July 2013

Background

Intra-specific variation in melanocyte pigmentation, common in the animal kingdom, has caught the eye of naturalists and biologists for centuries. In vertebrates, dark, eumelanin pigmentation is often genetically determined and associated with various behavioral and physiological traits, suggesting that the genes involved in melanism have far reaching pleiotropic effects. The mechanisms linking these traits remain poorly understood, and the potential involvement of developmental processes occurring in the brain early in life has not been investigated. We examined the ontogeny of rapid eye movement (REM) sleep, a state involved in brain development, in a wild population of barn owls (Tyto alba) exhibiting inter-individual variation in melanism and covarying traits. In addition to sleep, we measured melanistic feather spots and the expression of a gene in the feather follicles implicated in melanism (PCSK2).

Results

As in mammals, REM sleep declined with age across a period of brain development in owlets. In addition, inter-individual variation in REM sleep around this developmental trajectory was predicted by variation inPCSK2expression in the feather follicles, with individuals expressing higher levels exhibiting a more precocial pattern characterized by less REM sleep. Finally,PCSK2expression was positively correlated with feather spotting.

Conclusions

We demonstrate that the pace of brain development, as reflected in age-related changes in REM sleep, covaries with the peripheral activation of the melanocortin system. Given its role in brain development, variation in nestling REM sleep may lead to variation in adult brain organization, and thereby contribute to the behavioral and physiological differences observed between adults expressing different degrees of melanism.
An improved 3D tetraculture system mimicking the cellular organisation at the alveolar barrier to study the potential toxic effects of particles on the lung
Authors: Klein Sebastian G , Serchi Tommaso , Hoffmann Lucien , Blömeke Brunhilde , Gutleb Arno C
Published: July 2013

Background

Exposure to fine and ultra-fine ambient particles is still a problem of concern in many industrialised parts of the world and the intensified use of nanotechnology may further increase exposure to small particles. Complex in vitro coculture systems may be valuable tools to study particle-induced processes and to extrapolate effects of particles on the lung. A system consisting of four different human cell lines which mimics the cell response of the alveolar surface in vitro was developed to study native aerosol exposure (Vitrocell™ chamber). The system is composed of an alveolar type-II cell line (A549), differentiated macrophage-like cells (THP-1), mast cells (HMC-1) and endothelial cells (EA.hy 926), seeded in a 3D-orientation on a microporous membrane.

Results

The spatial distribution of the cells in the tetraculture was analysed by confocal laser scanning microscopy (CLSM), showing a confluent layer of endothelial and epithelial cells on both sides of the transwell. Macrophage-like cells and mast cells can be found on top of the epithelial cells. The cells formed colonies under submerged conditions, which disappeared at the ALI. To evaluate the response to oxidative stress, the dichlorodihydrofluorescein diacetate (DCFH-DA) assay was used together with 2,2’-azobis-2-methyl-propanimidamide-dihydrochloride (AAPH) as inducer of oxidative stress. The tetraculture showed less induction of reactive oxygen species (ROS) production after being treated with a positive control compared to the monocultures of EA.hy 926, THP-1 and HMC-1. Submerged cultures showed elevated ROS and IL-8 levels compared to ALI cultures. The Vitrocell™ aerosol exposure system was not significantly influencing the viability. Using this system, cells were exposed to an aerosol of 50 nm SiO2-Rhodamine NPs in PBS. The distribution of the NPs in the tetraculture after exposure was evaluated by CLSM. Fluorescence from internalized particles was detected in CD11b-positive THP-1 cells only.

Conclusion

The system can be used in conjunction with a native aerosol exposure system and may finally lead to a more realistic judgement regarding the hazard of new compounds and/or new nano-scaled materials in the future. The results for the ROS production and IL-8 secretion suggest that submerged exposure may lead to an overestimation of observed effects.
Green synthesis of a typical chiral stationary phase of cellulose-tris(3, 5-dimethylphenylcarbamate)
Authors: Liu Run-Qiang , Bai Lian-yang , Zhang Yi-Jun , Zhang Yu-Ping
Published: July 2013

Background

At present, the study on the homogeneous-phase derivatization of cellulose in ionic liquid is mainly focused on its acetylation. To the best of our knowledge, there has been no such report on the preparation of cellulose-tris(3,5-dimethylphenylcarbamate) (CDMPC) with ionic liquid 1-allyl-3-methyl-imidazolium chloride (AmimCl) so far.

Results

With ionic liquid 1-allyl-3-methylimidazolium chloride (AmimCl) as a reaction solvent, cellulose-tris(3,5-dimethylphenylcarbamate) (CDMPC) was synthesized by the reaction of 3,5-dimethylphenyl isocyanate and soluble microcrystalline cellulose in a homogeneous phase. The synthesized CDMPC was then coated onto the surfaces of aminopropyl silica gel to prepare a chiral stationary phase (CSP). The prepared CSP was successfully used in chiral separation of seven racemic pesticides by high performance liquid chromatography (HPLC). Good chiral separation was obtained using n-hexane and different modifiers as the mobile phases under the optimal percentage and column temperature, with the resolution of metalaxyl, diniconazole, flutriafol, paclobutrazol, hexaconazole, myclobutanil and hexythiazox of 1.73, 1.56, 1.26, 1.00, 1.18, 1.14 and 1.51, respectively. The experimental results suggested it was a good choice using a green solvent of AmimCl for cellulose functionalization.

Conclusion

CDMPC was successfully synthesized as the chiral selector by reacting 3, 5-dimethylphenyl isocyanate with dissolved microcrystalline cellulose in a green ionic liquid of AmimCl.
Ventricular tachycardia – an atypical initial presentation of sarcoidosis: a case report
Authors: Ekka Meera , Sinha Sanjeev , Purushothaman Raghunandan , Naik Nitish , Narang Rajiv , Singh Lavleen
Published: July 2013

Introduction

Symptomatic cardiac involvement is seen in less than 5% of all cases of sarcoidosis. Although clinically apparent cardiac sarcoidosis is an uncommon entity, ventricular tachyarrhythmias as the first presenting symptom are very rare.

Case presentation

We discuss the case of a 41-year-old Asian woman who presented to our hospital with intermittent palpitation and on evaluation was diagnosed to have systemic sarcoidosis with cardiac involvement. She was started on multiple antiarrhythmic drugs and corticosteroids without any satisfactory response.

Conclusions

Our case report indicates that sarcoidosis can manifest as ventricular tachycardia without any detectable systemic findings. This makes sarcoidosis an important diagnostic consideration in patients with ventricular tachycardia of unknown origin given the high mortality associated with ventricular tachyarrhythmias.
How useful are child death reviews: a local area’s perspective
Authors: Mazzola Francesca , Mohiddin Abdu , Ward Malcolm , Holdsworth Gillian
Published: July 2013

Background

Child Death Overview Panels (CDOP) provide a multidisciplinary and confidential forum to learn from and reduce deaths in those under 18 years. How well they perform and how to improve their effectiveness is a question posed at both local and national levels in England. With this in mind, this study looked at the child death review process in two London boroughs with a joint CDOP.

Findings

Data on cases reviewed from April 2008 to January 2011 were analysed focusing on cause of death and modifiable factors. Key stakeholders involved in the child death review process were interviewed regarding the effectiveness of the local death review process with responses analysed thematically.

105 (50.5%) of all notified deaths were reviewed to completion by CDOP of which 26.7% had modifiable factors. Neonates were the largest group of deaths (42.8%). Stakeholders found reviews time consuming, required significant administration and better integration with related processes e.g. hospital mortality meetings. Too much time was spent analysing cases of limited modifiability such as neonates. Implementation of recommendations needed strengthening and inclusion into the wider health and social care economy including joint strategic needs assessments and commissioning processes. Delayed reporting of information on cases contributed to a backlog.

Conclusions

The current process is bureaucratic, should better address neonatal deaths and needs more focus on implementing recommendations. Solutions include simpler forms, neonates-only subgroups, and linking recommendations to strategic initiatives such as Health and Wellbeing Boards.
Prevalence of malocclusions, oral habits and orthodontic treatment need in a 7- to 15-year-old schoolchildren population in Tirana
Authors: Laganà Giuseppina , Masucci Caterina , Fabi Francesco , Bollero Patrizio , Cozza Paola
Published: July 2013

Background

The aim of present study was to determine the prevalence of malocclusions, oral habits and the need for orthodontic treatment in a sample of 7- to 15-year-old Albanese schoolchildren.

Methods

The final sample comprised 2,617 subjects (1,257 males and 1,360 females), all orthodontically untreated. Occlusal relationship and the functional analysis were recorded for all subjects. The prevalence rates for the dental health component of the index of orthodontic treatment need (IOTN) were calculated. Comparisons between genders were performed for the prevalence of malocclusions, oral habits and IOTN grades (chi-square tests).

Results

Class I, class II and class III malocclusions and asymmetries were observed in 40.4%, 29.2%, 3.2% and 27.1% of the sample, respectively. There were 2,108 subjects (80.6%) that showed oral habits, with females (82.1%) presenting with a greater prevalence rate than males (78.9%). The objective need for orthodontic treatment (grades 4 and 5 of IOTN) was registered in 1,077 subjects (41.2%). This prevalence rate is higher than those reported for other European countries. No significant differences between genders were found for the IOTN grades.

Conclusions

The findings of the present study revealed the need to improve public health plans for orthodontic prevention and screening and to organise the resources in this area in Albania.
The immunogenicity to the first anti-TNF therapy determines the outcome of switching to a second anti-TNF therapy in spondyloarthritis patients
Authors: Plasencia Chamaida , Pascual-Salcedo Dora , García-Carazo Sara , Lojo Leticia , Nuño Laura , Villalba Alejandro , Peiteado Diana , Arribas Florencia , Díez Jesus , López-Casla Maria Teresa , Martín-Mola Emilio , Balsa Alejandro
Published: July 2013

Introduction

Anti-TNF drugs have proven to be effective against spondyloarthritis (SpA), although 30% of patients fail to respond or experience adverse events leading to treatment discontinuation. In rheumatoid arthritis, the presence of anti-drug antibodies (ADA) against the first TNF inhibitor influences the outcome after switching. Our aim was to assess whether the response to a second anti-TNF drug is related to the previous development of ADA to the first anti-TNF drug SpA patients.

Methods

Forty-two SpA patients began a second anti-TNF drug after failing to respond to the first anti-TNF therapy. Clinical activity was assessed by the Ankylosing Spondylitis Disease Activity Score (ASDAS) at baseline (at the beginning of the first and second anti-TNF therapy) and at 6 months after switching. The drug and ADA levels were measured by ELISA before each administration.

Results

All patients were treated with anti-TNF drugs and mainly due to inefficacy were switched to a second anti-TNF drug. Eleven of 42 (26.2%) developed ADA during the first biologic treatment. At baseline, no differences in ASDAS were found in patients with or without ADA to the first anti-TNF drug (3.52 ± 1.03 without ADA vs. 3.14 ± 0.95 with ADA, p = 0.399) and to the second anti-TNF drug (3.36 ± 0.94 without ADA vs. 3.09 ± 0.91 with ADA, p = 0.466). At 6 months after switching, patients with previous ADA had lower disease activity (1.62 ± 0.93 with ADA vs. 2.79 ± 1.01 without ADA, p = 0.002) and most patients without ADA had high disease activity state by the ASDAS (25 out of 31 (80.6%) without ADA vs. 3 out of 11 (27.3%) with ADA, p = 0.002).

Conclusions

In SpA the failure to respond to the first anti-TNF drug due to the presence of ADA predicts a better clinical response to a second anti-TNF drug.
MicroRNA-21 promotes the ovarian teratocarcinoma PA1 cell line by sustaining cancer stem/progenitor populations in vitro
Authors: Chung Wei-Min , Chang Wei-Chun , Chen Lumin , Chang Ying-Yi , Shyr Chih-Rong , Hung Yao-Ching , Ma Wen-Lung
Published: July 2013

Introduction

Resistance of cancer stem/progenitor cells (CSPCs) to chemotherapy can lead to cancer relapse. Ovarian teratocarcinoma (OVTC) arises from germ cells and comprises pluripotent cells that can be used to study cancer cell stemness. In this study, we evaluated whether microRNA-21 (miR-21) promotes ovarian teratocarcinoma by maintaining cancer stem/progenitor populations.

Methods

The lentiviral delivery system was used to upregulate or to suppress the expression of miR-21 in the human ovarian teratocarcinoma cell line PA1 and cell growth assays were used to monitor the expression of miR-21 at different time points. Antibodies directed toward CD133, a stem cell marker, were used to identify CSPCs in the PA1 cell population, and the level of miR-21 expression was determined in enriched CSPCs. Stem cell functional assays (sphere assay and assays for CD133 expression) were used to assess the effects of miR-21 on progression of the CD133+ population.

Results

Knockdown of miR-21 in PA1 cells attenuated growth of PA1 cells whereas overexpression of miR-21 promoted cell growth. Moreover, knockdown of miR-21 resulted in a marked reduction in the CD133+ population and sphere formation of CSPCs. In contrast, overexpression of miR-21 resulted in a marked increase in the population of CD133+ cells as well as sphere formation of CSPCs.

Conclusions

MicroRNA-21 plays a significant role in cancer growth by regulating stemness in cancer cells.
α-arrestin 1 (ARRDC1) and β-arrestins cooperate to mediate Notch degradation in mammals
Authors: Puca Loredana , Chastagner Patricia , Meas-Yedid Vannary , Israël Alain , Brou Christel
Published: July 2013

Notch signaling is a conserved signaling pathway implicated in embryogenesis and adult tissue maintenance. Notch signaling strength is strictly regulated, notably by maintaining a controlled pool of functional receptor at the cell surface. Mammalian non-activated Notch receptor is internalized, ubiquitylated by the Itch E3 ubiquitin ligase and degraded in the lysosomes. Here, we show that β-arrestins are necessary for Itch–Notch interaction and for Itch-driven ubiquitylation and degradation of Notch. Interestingly, β-arrestins do not directly bind Itch but heterodimerize with a member of another subfamily of arrestins called ARRDC1 or α-arrestin 1, which harbors PPxY motifs that allow direct interaction with Itch. Cells transfected with ARRDC1 mutated in PPxY motifs show reduced Itch-mediated Notch ubiquitylation and impaired lysosomal degradation of Notch, as observed in β-arrestin−/− or Itch−/− cells. Our data show for the first time that ARRDC1 and β-arrestins heterodimerize and cooperate in the same complex to promote non-activated Notch receptor degradation, thus acting as negative regulators of Notch signaling.
Ras activation and symmetry breaking duringDictyosteliumchemotaxis
Authors: Kortholt Arjan , Keizer-Gunnink Ineke , Kataria Rama , Van Haastert Peter J. M.
Published: July 2013

Central to chemotaxis is the molecular mechanism by which a shallow spatial gradient of chemoattractant induces symmetry breaking of activated signaling molecules. Previously, we have used Dictyostelium mutants to investigate the minimal requirements for chemotaxis, and identified a basal signaling module providing activation of Ras and F-actin at the leading edge. Here, we show that Ras activation after application of a pipette releasing the chemoattractant cAMP has three phases, each depending on specific guanine-nucleotide-exchange factors (GEFs). Initially a transient activation of Ras occurs at the entire cell boundary, which is proportional to the local cAMP concentrations and therefore slightly stronger at the front than in the rear of the cell. This transient Ras activation is present in gα2 (gpbB)-null cells but not in gβ (gpbA)-null cells, suggesting that Gβγ mediates the initial activation of Ras. The second phase is symmetry breaking: Ras is activated only at the side of the cell closest to the pipette. Symmetry breaking absolutely requires Gα2 and Gβγ, but not the cytoskeleton or four cAMP-induced signaling pathways, those dependent on phosphatidylinositol (3,4,5)-triphosphate [PtdIns(3,4,5)P3], cGMP, TorC2 and PLA2. As cells move in the gradient, the crescent of activated Ras in the front half of the cell becomes confined to a small area at the utmost front of the cell. Confinement of Ras activation leads to cell polarization, and depends on cGMP formation, myosin and F-actin. The experiments show that activation, symmetry breaking and confinement of Ras during Dictyostelium chemotaxis uses different G-protein subunits and a multitude of Ras GEFs and GTPase-activating proteins (GAPs).
Development of Ruthenium-loaded Alkaline-earth Titanates as Catalysts for Ammonia Synthesis
Authors: Horiuchi Yu , Kamei Genki , Saito Masakazu , Matsuoka Masaya
Published: July 2013

Ru-loaded alkaline-earth titanates show efficient catalytic activities for ammonia synthesis because of the electronic promotion effects induced by alkaline-earth elements and the low acidity of the support materials. Among those explored, the best catalytic performance was obtained by using barium titanate as the support material.
Computational detection of abundant long-range nucleotide covariation inDrosophilagenomes
Authors: Bindewald Eckart , Shapiro Bruce A.
Published: July 2013

Functionally important nucleotide base-pairing often manifests itself in sequence alignments in the form of compensatory base changes (covariation). We developed a novel index-based computational method (CovaRNA) to detect long-range covariation on a genomic scale, as well as another computational method (CovStat) for determining the statistical significance of observed covariation patterns in alignment pairs. Here we present an all-versus-all search for nucleotide covariation inDrosophilagenomic alignments. The search is genome wide, with the restriction that only alignments that correspond to euchromatic regions, which consist of at least 10Drosophilaspecies, are being considered (59% of the euchromatic genome ofDrosophila melanogaster). We find that long-range covariations are especially prevalent between exons of mRNAs as well as noncoding RNAs; the majority of the observed covariations appear as not reverse complementary, but as synchronized mutations, which could be due to interactions with common interaction partners or due to the involvement of genomic elements that are antisense of annotated transcripts. The involved genes are enriched for functions related to regionalization as well as neural and developmental processes. These results are computational evidence that RNA–RNA long-range interactions are a widespread phenomenon that is of fundamental importance to a variety of cellular processes.
2-Selenouridine triphosphate synthesis and Se-RNA transcription
Authors: Sun Huiyan , Jiang Sibo , Caton-Williams Julianne , Liu Hehua , Huang Zhen
Published: July 2013

2-Selenouridine (SeU) is one of the naturally occurring modifications of Se-tRNAs (SeU-RNA) at the wobble position of the anticodon loop. Its role in the RNA-RNA interaction, especially during the mRNA decoding, is elusive. To assist the research exploration, herein we report the enzymatic synthesis of the SeU-RNA via 2-selenouridine triphosphate (SeUTP) synthesis and RNA transcription. Moreover, we have demonstrated that the synthesized SeUTP is stable and recognizable by T7 RNA polymerase. Under the optimized conditions, the transcription yield of SeU-RNA can reach up to 85% of the corresponding native RNA. Furthermore, the transcribed SeU-hammerhead ribozyme has the similar activity as the corresponding native, which suggests usefulness of SeU-RNAs in function and structure studies of noncoding RNAs, including the Se-tRNAs.
RNase III is required for localization to the nucleoid of the 5′ pre-rRNA leader and for optimal induction of rRNA synthesis inE. coli
Authors: Malagon Francisco
Published: July 2013

It has recently been demonstrated that ribosomes are preferentially localized outside the nucleoid inEscherichia coli, but little is known about the spatial regulation of pre-rRNA processing. In this work, I investigate the cellular distribution of leader pre-rRNAs using RNA-FISH. In contrast to mature rRNA, the 5′ proximal leader region associates with the nucleoid, and this association occurs in an RNase III-dependent manner. Moreover, RNase III plays a role in the rapid induction of ribosomal operons during outgrowth and is essential in the absence of the transcriptional regulator Fis, suggesting a linkage of transcription and RNA processing for ribosomal operons inE. coli.