bluuebunny/crossref_metadata_2025_split huggingface dataset:
Validation of microinjection methods for generating knockout mice by CRISPR/Cas-mediated genome engineering
Authors: Horii Takuro , Arai Yuji , Yamazaki Miho , Morita Sumiyo , Kimura Mika , Itoh Masahiro , Abe Yumiko , Hatada Izuho 
Published: March 2014

AbstractThe CRISPR/Cas system, in which the Cas9 endonuclease and a guide RNA complementary to the target are sufficient for RNA-guided cleavage of the target DNA, is a powerful new approach recently developed for targeted gene disruption in various animal models. However, there is little verification of microinjection methods for generating knockout mice using this approach. Here, we report the verification of microinjection methods of the CRISPR/Cas system. We compared three methods for injection: (1) injection of DNA into the pronucleus, (2) injection of RNA into the pronucleus and (3) injection of RNA into the cytoplasm. We found that injection of RNA into the cytoplasm was the most efficient method in terms of the numbers of viable blastocyst stage embryos and full-term pups generated. This method also showed the best overall knockout efficiency.
Metabolic gene profile in early human fetal heart development
Authors: Iruretagoyena J.I. , Davis W. , Bird C. , Olsen J. , Radue R. , Teo Broman A. , Kendziorski C. , Splinter BonDurant S. , Golos T. , Bird I. , Shah D. 
Published: March 2014

Abstract
The primitive cardiac tube starts beating 6–8 weeks post fertilization in the developing embryo. In order to describe normal cardiac development during late first and early second trimester in human fetuses this study used microarray and pathways analysis and created a corresponding ‘normal’ database. Fourteen fetal hearts from human fetuses between 10 and 18 weeks of gestational age (GA) were prospectively collected at the time of elective termination of pregnancy. RNA from recovered tissues was used for transcriptome analysis with Affymetrix 1.0 ST microarray chip. From the amassed data we investigated differences in cardiac development within the 10–18 GA period dividing the sample by GA in three groups: 10–12 (H1), 13–15 (H2) and 16–18 (H3) weeks. A fold change of 2 or above adjusted for a false discovery rate of 5% was used as initial cutoff to determine differential gene expression for individual genes. Test for enrichment to identify functional groups was carried out using the Gene Ontology (GO) and the Kyoto Encyclopedia of Genes and Genomes (KEGG). Array analysis correctly identified the cardiac specific genes, and transcripts reported to be differentially expressed were confirmed by qRT–PCR. Single transcript and Ontology analysis showed first trimester heart expression of myosin-related genes to be up-regulated &gt;5-fold compared with second trimester heart. In contrast the second trimester hearts showed further gestation-related increases in many genes involved in energy production and cardiac remodeling. In conclusion, fetal heart development during the first trimester was dominated by heart-specific genes coding for myocardial development and differentiation. During the second trimester, transcripts related to energy generation and cardiomyocyte communication for contractile coordination/proliferation were more dominant. Transcripts related to fatty acid metabolism can be seen as early as 10 weeks and clearly increase as the heart matures. Retinol receptor and gamma-aminobutyric acid (GABA) receptor transcripts were detected, and have not been described previously in human fetal heart during this period. For the first time global gene expression of heart has been described in human samples to create a database of normal development to understand and compare with known abnormal fetal heart development.
Policy making, adult safeguarding and public health: a formula for change?
Authors: Quigley Leo 
Published: March 2014

Purpose
– The purpose of this paper is to review the reasons underlying the slow rate of progress towards developing a comprehensive policy underpinning for adult safeguarding in England and proposes long-term solutions.

Design/methodology/approach
– This paper uses a model of policy change to argue that adult safeguarding has been over-reliant on case histories to define its policy problems and influence its politics, while making insufficient progress on data collection and analysis. It uses examples from the parallel discipline of public health to explore four challenges, or “problems”, relevant to the further development of the knowledge base underpinning adult safeguarding policy.

Findings
– Four recommendations emerge for closing the adult safeguarding “knowledge gap”, including the development of a national research strategy for adult safeguarding. In a fifth recommendation the paper also proposes a clearer recognition of the contribution that local public health professionals can make to local adult safeguarding policy making and programme development.

Practical implications
– The first four recommendations of this paper would serve as the basis for developing a national research strategy for adult safeguarding. The fifth would strengthen the contribution of local public health departments to safeguarding adults boards.

Originality/value
– The author is unaware of the existence of any other review of the limitations of the adult safeguarding knowledge base as a foundation for policy making, or which proposes strategic solutions. The work is valuable for its practical proposals.
How healthcare organisations can act as institutional entrepreneurs in a context of change
Authors: Breton Mylaine , Lamothe Lise , Denis Jean-Louis 
Published: March 2014

Purpose– The aim of this paper is to illustrate and discuss how healthcare organisations can act as institutional entrepreneurs in a context of change.Design/methodology/approach– The authors conducted an in-depth longitudinal case study (2005-2008) of a healthcare organisation in the province of Quebec, Canada. Data collection consisted of real-time observations of senior managers (n=87), interviews (n=24) with decision-makers and secondary data analysis of documents.Findings– The paper reports on the extent to which entrepreneurial healthcare organisations can be a driving force in the creation of a new practice. The authors analyse the development of a diabetes reference centre by a healthcare organisation acting as an institutional entrepreneur that illustrates the conceptualisation of an innovation and the mobilisation of resources to implement it and to influence other actors in the field. The authors discuss the case in reference to three stages of change: emergence, implementation and diffusion. The results illustrate the different strategies used by managers to advance their proposed projects.Research limitations/implications– This study helps to better understand the dynamics of mandated change in a mature field such as healthcare and the roles played by organisations in this process. By adopting a proactive strategy, a healthcare organisation can play an active role and strongly influence the evolution of its field.Originality/value– This paper is one of only a few to analyse strategies used by healthcare organisations in the context of mandated change.
Association ofHMGB1andHMGB2genetic polymorphisms with lung cancer chemotherapy response
Authors: Wang Ying , Li Xiang‐Ping , Yin Ji‐Ye , Zhang Yu , He Hui , Qian Chen‐Yue , Chen Juan , Zheng Yi , Smieszkol Kamila , Fu Yi‐Lan , Chen Zi‐Yu , Zhou Hong‐Hao , Liu Zhao‐Qian 
Published: March 2014

SummaryThe aim of the present study was to investigate the association of genetic polymorphisms in high mobility group box 1 and 2 (HMGB1andHMGB2, respectively) with platinum‐based chemotherapy responses inChinese lung cancer patients. In total, 338 Chinese lung cancer patients (154 responders and 184 non‐responders) were recruited to the study. All patients received at least two cycles of first‐line platinum‐based chemotherapy. Three tagging single nucleotide polymorphisms (SNPs) ofHMGB1and two taggingSNPs ofHMGB2were detected in patients. We found that rs1412125 and rs2249825 ofHMGB1were significantly associated with the platinum‐based chemotherapy response in both recessive and genotypic models. In addition, rs1412125 showed significant association with platinum‐based chemotherapy response for the subgroup of patients aged >55 years in additive, recessive and genotypic models. No significant associations were detected between otherSNPs and the platinum‐based chemotherapy response. TheHMGB1SNPs (rs1412125 and rs2249825) were associated with platinum‐based chemotherapy responses in Chinese lung cancer patients. In conclusion,HMGB1SNPs may serve as potential biomarkers for predicting the efficacy of platinum‐based chemotherapy.
Association of nitric oxide synthase 3 (NOS3) 894 G>T polymorphism with prognostic outcomes of anthracycline in Chinese patients with acute myeloid leukaemia
Authors: He Hui , Xu Ya‐Jing , Yin Ji‐Ye , Li Xi , Qu Jian , Xu Xiao‐Jing , Liu Zhuo‐Gang , Zhou Fan , Zhai Ming , Li Yan , Zhou Hong‐Hao , Liu Zhao‐Qian 
Published: March 2014

SummaryThe aim of the present study was to investigate the influence of the nitric oxide synthase 3 (NOS3) 894 G>T polymorphism on prognostic outcomes of anthracycline in Chinese patients with de novo intermediate‐risk acute myeloid leukaemia (AML) and to examine the gene expression level in relation to genetic variation. In all, 225 Chinese patients with intermediate‐risk AML (at the complete remission stage) treated with anthracycline were enrolled in the study. The 894 G>T polymorphism of the NOS3 gene was analysed by allele‐specific matrix‐assisted laser desorption ionization time‐of‐flight. Expression of NOS3 mRNA was tested in 72 patients of known genotype for NOS3 894 G>T. The clinical characteristics of these patients were obtained from medical records. Survival analysis showed that patients with AML (GG genotype) had a longer overall survival (OS; P = 0.006). After adjusting for age, gender, leucocyte count, haemoglobin level, platelet level, French, American and Britain (FAB) classification, lactate dehydrogenase levels, Eastern Cooperative Oncology Group Performance Status, nucleophosmin gene and fms‐related tyrosine kinase 3 gene, multivariate survival analysis showed that the NOS3 894 G>T polymorphism appeared to be a predicting factor for OS (P = 0.014; hazard ratio = 1.856). However, no significant associations between the NOS3 894 G>T polymorphism and relapse‐free survival and relapse in patients with AML were observed. Gene expression levels were significantly higher in patients with the GG genotype than in patients with the GT and TT genotypes (P = 0.033). The findings suggest that the NOS3 894 G>T variant may be a biomarker for the prediction of OS in Chinese patients with AML.
Functional coexistence of twin arsenic resistance systems in Pseudomonas putida KT2440
Authors: Páez‐Espino A. David , Durante‐Rodríguez Gonzalo , de Lorenzo Víctor 
Published: March 2014

SummaryThe genome of the soil bacterium Pseudomonas putida KT2440 bears two virtually identical arsRBCH operons putatively encoding resistance to inorganic arsenic species. Single and double chromosomal deletions in each of these ars clusters of this bacterium were tested for arsenic sensitivity and found that the contribution of each operon to the resistance to the metalloid was not additive, as either cluster sufficed to endow cells with high‐level resistance. However, otherwise identical traits linked to each of the ars sites diverged when temperature was decreased. Growth of the various mutants at 15°C (instead of the standard 30°C for P. putida) uncovered that ars2 affords a much higher resistance to As (III) than the ars1 counterpart. Reverse transcription polymerase chain reaction of arsB1 and arsB2 genes as well as lacZ fusions to the Pars1 and Pars2 promoters traced the difference to variations in transcription of the corresponding gene sets at each temperature. Functional redundancy may thus be selected as a stable condition – rather than just as transient state – if it affords one key activity to be expressed under a wider range of physicochemical settings. This seems to provide a straightforward solution to regulatory problems in environmental bacteria that thrive under changing scenarios.
Analysts' forecast error: a robust prediction model and its short‐term trading profitability
Authors: Boudt Kris , de Goeij Peter , Thewissen James , Van Campenhout Geert 
Published: March 2014

AbstractThis paper contributes to the empirical evidence on the investment horizon salient to trading based on predicting the error in analysts' earnings forecasts. An econometric framework is proposed that accommodates the stylized fact of extreme values in the forecast error series. We find that between 1998 and 2010, the strategy of taking a long (short) position in stocks with the most pessimistic (optimistic) I/B/E/S forecast has an annual risk‐adjusted return of 16.56 per cent before transaction costs. The robust method used to predict this pessimism (optimism) and the one‐week investment horizon are the key drivers of the strategy's profitability.
Genetic variants in KDR transcriptional regulatory region affect promoter activity and intramuscular fat deposition in Erhualian pigs
Authors: Fu Y. , Sun W. , Xu C. , Gu S. , Li Y. , Liu Z. , Chen J. 
Published: March 2014

SummaryKinase insert domain receptor (KDR), a vascular endothelial growth factor (VEGF) receptor, is widely regarded as having a principal role in mediating VEGF‐induced responses in angiogenesis. As angiogenesis provides oxygen and nutrients for growth and deposition of adipose cells, our objective was to determine whether the promoter polymorphisms in the KDR gene have effects on intramuscular fat (IMF) deposition in the longissimus dorsi muscle. Three novel SNPs, c.‐1316A>G, c.‐1303C>T and c.‐1108A>C, were revealed to have differential allele distribution between high‐ and low‐IMF content groups by comparative sequencing of DNA pools. The three SNPs were completely linked, forming only ACA or GTC haplotypes when genotyped in 105 Erhualian purebred pigs and 98 Duroc × Large White × Yorkshire (D×L×Y) cross‐bred pigs. It is interesting that the ACA haplotype is present exclusively in Erhualian pigs and not in D×L×Y pigs. The ACA promoter was found to have higher activity than GTC type for KDR transcription using either gene expression analysis or luciferase assay. Site‐direct mutation analysis demonstrated that c.‐1316A>G is the causation of promoter activity alteration. Furthermore, we detected that CD31 (also known as PECAM1) and CD34, two blood vessel endothelial markers, expressed higher in ACA/ACA individuals. We concluded that the ACA promoter might be a desirable form for improving IMF content by promoting higher KDR gene expression and more blood vessel network.
Thyroidal abscesses in third and fourth branchial anomalies: not only a paediatric diagnosis
Authors: Kruijff Schelto , Sywak Mark S. , Sidhu Stan B. , Shun Albert , Novakovic Daniel , Lee James C. , Delbridge Leigh W. 
Published: March 2014

AbstractIntroductionAcute suppurative thyroiditis and recurrent abscess formation due to third and fourth branchial anomalies typically present in children. However, thyroid abscesses in branchial anomalies may occur in adulthood as well. Failure to recognize and delayed drainage of a neck abscess may lead to a fulminant life‐threatening outcome.MethodsThis is a retrospective case series. The study group comprised all patients presenting over a 12‐month period from January to December 2012 with thyroid abscesses and a branchial cleft anomaly in two centres, one adult and the other paediatric. Patient demographics, clinical presentation, imaging, surgical management, definitive histology and outcomes were documented.ResultsFive patients were identified with a history of thyroid abscesses. Only one was a child (aged 9 years) with the other four being adults (aged 20, 34, 37 and 41 years). All patients had third or fourth left branchial cleft anomalies, presenting as suppurative thyroiditis with a left‐sided thyroid abscess. Management options ranged from abscess drainage on initial presentation, primary thyroid lobectomy or delayed thyroid lobectomy following abscess drainage.ConclusionAcute suppurative thyroidits and thyroid abscesses is not just a paediatric diagnosis but may present at any age. In both children and adults, a thyroid abscess almost always arises from branchial cleft anomalies.
The progression of myopia from its onset at age 8–12 to adulthood and the influence of heredity and external factors on myopic progression. A 23‐year follow‐up study
Authors: Pärssinen Olavi , Kauppinen Markku , Viljanen Anne 
Published: March 2014

AbstractPurposeTo examine myopic progression and factors connected with myopic progression.MethodsMyopic schoolchildren, with no previous spectacles, 119 boys and 121 girls, were recruited during 1983–1984 to a randomized 3‐year clinical trial of bifocal treatment of myopia with a subsequent 20‐year follow‐up. Participants' mean age at Baseline was 10.9, ranging from 8.7 to 12.8 years. An ophthalmological examination was carried out annually for 3 years and twice thereafter at ca. 10‐year intervals. Additional refraction values were received from prescriptions issued by different ophthalmologists and opticians. Altogether, 1915 refraction values were available. Reading distance and accommodation were measured at each control visit. Data on parents' myopia, daily time spent on reading and close work, outdoor activities and watching television were gathered with a structured questionnaire.ResultsUsing bifocals (+1.75 add) or reading without glasses or accommodation stimulus during the 3‐year period in childhood did not correlate with adulthood refraction. Short reading distance in childhood predicted higher adulthood myopia among females. The factors predicting faster myopic progression were parents' myopia and less time spent on sports and outdoor activities at childhood. Time spent on reading and close work in childhood was related to myopic progression during the first 3 years but did not predict adulthood myopia. Myopia throughout follow‐up was higher among those who watched television <3 hr daily than those who spent more time watching television. Mean myopic progression 8 years after age 20–24 was −0.45 D ± 0.71 (SD), and in 45% of cases, progression was ≥0.5 D.ConclusionsIn nearly half of the cases, myopia beginning at school continued to progress into adulthood. Higher adulthood myopia was mainly related to parents' myopia and less time spent on sports and outdoor activities in childhood.
LAWYERS IN VENTURE CAPITAL CONTRACTING: THEORY AND EVIDENCE
Authors: BENGTSSON OLA , BERNHARDT DAN 
Published: March 2014

AbstractReal‐world financial contracts are sometimes so complex that it can be difficult to understand their exact payoff consequences. We develop and test a theoretical model of a venture capitalist (VC) negotiating with an entrepreneur who may overweigh or underweigh the payoff consequences of contractual downside protection (DP). A lawyer with expertise in venture capital can inform the entrepreneur about these consequences, but less expert (but otherwise high quality) lawyers cannot. We determine how a VC's decision to include DP is affected by the expected quality of the entrepreneur's project, the entrepreneur's experience, and the VC expertise of his/her legal counsel. We show that the VC's incentive to include unnecessary DP declines in expected project quality. Indeed, for inexperienced entrepreneurs involved with high‐quality projects, VCs prefer that the entrepreneur's counsel has VC expertise. This implies that, when negotiating with inexperienced entrepreneurs, VCs who invest in high‐quality companies should be more likely to negotiate with entrepreneurs who employ lawyers with VC expertise. We document broad empirical support for the model, and provide evidence against competing explanations. (JEL L24, G24, K12, L14, L24)
Integrative diagnosis of the needle nematodeLongidorus jonesiaffecting forest in southernJapan
Authors: Palomares‐Rius J. E. , Kikuchi T. , Cantalapiedra‐Navarrete C. , Castillo P. 
Published: March 2014

SummaryThe occurrence of a parthenogenetic population ofLongidorus jonesiin the rhizosphere of aPrunussp. tree in a mixed evergreen forest is reported for the first time inMiyazaki (Miyazaki prefecture), southernJapan, and constitutes the first report of this species inJapan. Nematode diagnosis matched well with the original description in India and a subsequent report inChina. Morphometric traits of theJapanese population were characterized by a longer odontostyle length (123–142.5vs. 107–120 μm) and some differences in c ratio (95.9–131.5vs. 140–185) with respect to the original description, but closer to theChinese population. This difference could be considered as geographical intraspecific variability. Molecular characterization from nuclear ribosomalRNAgenes [D2‐D3 expansion segments of large ribosomal subunit (28S) and internal transcribed spacer 1 (ITS1)] was provided for the first time. Molecular analysis ofD2‐D3 andITS1 showed a close relationship withL. diadecturus, a virus‐vector nematode, by 94 and 79% similarity, respectively. The similarities in morphology, molecular and ecology (L. jonesiandL. diadecturusare feeding mainly onPrunusspecies) suggest that this species could be a potential virus vector, as it is the case withL. diadecturusandpeach rosette mosaic virus.
High‐frequency p16INK4A promoter methylation is associated with histone methyltransferase SETDB1 expression in sporadic cutaneous melanoma
Authors: Kostaki Maria , Manona Argyro D. , Stavraka Irene , Korkolopoulou Penelope , Levidou Georgia , Trigka Eleni‐Andriana , Christofidou Eleftheria , Champsas Grigorios , Stratigos Alexandros J. , Katsambas Andreas , Papadopoulos Othon , Piperi Christina , Papavassiliou Athanasios G. 
Published: March 2014

AbstractEpigenetic mechanisms participate in melanoma development and progression. The effect of histone modifications and their catalysing enzymes over euchromatic promoter DNA methylation in melanoma remains unclear. This study investigated the potential association of p16INK4A promoter methylation with histone methyltransferase SETDB1 expression in Greek patients with sporadic melanoma and their correlation with clinicopathological characteristics. Promoter methylation was detected by methylation‐specific PCR in 100 peripheral blood samples and 58 melanoma tissues from the same patients. Cell proliferation (Ki‐67 index), p16INK4A and SETDB1 expression were evaluated by immunohistochemistry. High‐frequency promoter methylation (25.86%) was observed in tissue samples and correlated with increased cell proliferation (P = 0.0514). p16INK4A promoter methylation was higher in vertical growth‐phase (60%) melanomas than in radial (40%, P = 0.063) and those displaying epidermal involvement (P = 0.046). Importantly, p16INK4A methylation correlated with increased melanoma thickness according to Breslow index (P = 0.0495) and marginally with increased Clark level (I/II vs III/IV/V, P = 0.070). Low (1–30%) p16INK4A expression was detected at the majority (19 of 54) of melanoma cases (35.19%), being marginally correlated with tumor lymphocytic infiltration (P = 0.078). SETDB1 nuclear immunoreactivity was observed in 47 of 57 (82.46%) cases, whereas 27 of 57 (47.37%) showed cytoplasmic immunoexpression. Cytoplasmic SETDB1 expression correlated with higher frequency of p16INK4A methylation and p16INK4A expression (P = 0.033, P = 0.011, respectively). Increased nuclear SETDB1 levels were associated with higher mitotic count (0–5/mm2 vs >5/mm2, P = 0.0869), advanced Clark level (III‐V, P = 0.0380), epidermal involvement (P = 0.0331) and the non‐chronic sun exposure‐associated melanoma type (P = 0.0664). Our data demonstrate for the first time the association of histone methyltransferase SETDB1 with frequent methylation of the euchromatic p16INK4A promoter and several prognostic parameters in melanomas.