bluuebunny/crossref_metadata_2025_split huggingface dataset:
Organizational innovations and health care decentralization: a perspective from Spain
Authors: LÓPEZ-CASASANOVAS GUILLEM 
Published: July 2007

Abstract:Recent policy developments in public health care systems lead to a greater diversity in health care. Decentralization, either geographically or at an institutional level, is the key force, because it encourages innovation and local initiatives in health care provision. The devolution of responsibilities allows for a sort of ‘deconstruction’ of the status quo by changing both organizational forms and service provision. The new organizations enjoy greater freedom in the way they pay their staff, and are judged according to the results. These organizations may retain financial surpluses, develop ‘spin-off’ companies, and commission a range of specialized services (such as Diagnostic and Treatment Centres in UK) from providers outside the institutional setting in order to have more access to capital markets. However, this diversity may generate a feeling of a lack of commitment to a national health service and ultimately a loss of social cohesion. Through fiscal decentralization to regional authorities or planned delegation of financial agreements to providers, financial incentives are more explicit and may seem to place profit making above a commitment to better health care. An evaluation of the ‘myths and realities’ of the decentralization process is needed. Here, I offer an assessment of the ‘pros’ and ‘cons’ of the health care decentralization process in Spain, drawing on the experience of regional reforms from the pioneering organizational innovations implemented in Catalonia in 1981, up to the currently observed dispersion of health care spending per capita across the regions.
Supplementary health insurance as a tool for risk-selection in mandatory basic health insurance markets
Authors: PAOLUCCI FRANCESCO , SCHUT ERIK , BECK KONSTANTIN , GREß STEFAN , VAN DE VOORDE CARINE , ZMORA IRIT 
Published: July 2007

Abstract:As the share of supplementary health insurance (SI) in health care finance is likely to grow, SI may become an increasingly attractive tool for risk-selection in basic health insurance (BI). In this paper, we develop a conceptual framework to assess the probability that insurers will use SI for favourable risk-selection in BI. We apply our framework to five countries in which risk-selection via SI is feasible: Belgium, Germany, Israel, the Netherlands, and Switzerland. For each country, we review the available evidence of SI being used as selection device. We find that the probability that SI is and will be used for risk-selection substantially varies across countries. Finally, we discuss several strategies for policy makers to reduce the chance that SI will be used for risk-selection in BI markets.
Post‐ischemic treatment with erythropoietin or carbamylated erythropoietin reduces infarction and improves neurological outcome in a rat model of focal cerebral ischemia
Authors: Wang Y , Zhang Z G , Rhodes K , Renzi M , Zhang R L , Kapke A , Lu M , Pool C , Heavner G , Chopp M 
Published: July 2007

Background and purpose:Recombinant human erythropoietin (rhEPO; Epoetin‐α; PROCRIT™) has been shown to exert neuroprotective and restorative effects in a variety of CNS injury models. However, limited information is available regarding the dose levels required for these beneficial effects or the neuronal responses that may underlie them. Here we have investigated the dose‐response to rhEPO and compared the effects of rhEPO with those of carbamylated rhEPO (CEPO) in a model of cerebral stroke in rats.Experimental approach:Rats subjected to embolic middle cerebral artery occlusion (MCAo) were treated with rhEPO or CEPO, starting at 6 h and repeated at 24 and 48 h, after MCAo. Cerebral infarct volumes were assessed at 28 days and neurological impairment at 7, 14, 21 and 28 days, post‐MCAo.Key results:rhEPO at dose levels of 500, 1150 or 5000 IU kg−1 or CEPO at a dose level of 50 μg kg−1 significantly reduced cortical infarct volume and reduced neurologic impairment. All doses of rhEPO, but not CEPO, produced a transient increase in haematocrit, while rhEPO and CEPO substantially reduced the number of apoptotic cells and activated microglia in the ischemic boundary region.Conclusions and implications:These data indicate that rhEPO and CEPO have anti‐inflammatory and anti‐apoptotic effects, even with administration at 6 h following embolic MCAo in rats. Taken together, these actions of rhEPO and CEPO are likely to contribute to their reduction of neurologic impairment following cerebral ischemia.British Journal of Pharmacology (2007) 151, 1377–1384; doi:10.1038/sj.bjp.0707285
Specificity of the ecto‐ATPase inhibitor ARL 67156 on human and mouse ectonucleotidases
Authors: Lévesque S A , Lavoie É G , Lecka J , Bigonnesse F , Sévigny J 
Published: July 2007

Background and purpose:ARL 67156, 6‐N,N‐Diethyl‐D‐β‐γ‐dibromomethylene adenosine triphosphate, originally named FPL 67156, is the only commercially available inhibitor of ecto‐ATPases. Since the first report on this molecule, various ectonucleotidases responsible for the hydrolysis of ATP at the cell surface have been cloned and characterized. In this work, we identified the ectonucleotidases inhibited by ARL 67156.Experimental approach:The effect of ARL 67156 on recombinant NTPDase1, 2, 3 & 8 (mouse and human), NPP1, NPP3 and ecto‐5′‐nucleotidase (human) have been evaluated. The inhibition of the activity of NTPDases (using the following substrates: ATP, ADP, UTP), NPPs (pnp‐TMP, Ap3A) and ecto‐5′‐nucleotidase (AMP) was measured by colorimetric or HPLC assays.Key results:ARL 67156 was a weak competitive inhibitor of human NTPDase1, NTPDase3 and NPP1 with Ki of 11±3, 18±4 and 12±3 μM, respectively. At concentrations used in the literature (50–100 μM), ARL 67156 partially but significantly inhibited the mouse and human forms of these enzymes. NTPDase2, NTPDase8, NPP3 and ecto‐5′‐nucleotidase activities were less affected. Importantly, ARL 67156 was not hydrolysed by either human NTPDase1, 2, 3, 8, NPP1 or NPP3.Conclusions and implications:In cell environments where NTPDase1, NTPDase3, NPP1 or mouse NTPDase8 are present, ARL 67156 would prolong the effect of endogenously released ATP on P2 receptors. However, it does not block any ectonucleotidases efficiently when high concentrations of substrates are present, such as in biochemical, pharmacological or P2X7 assays. In addition, ARL 67156 is not an effective inhibitor of NTPDase2, human NTPDase8, NPP3 and ecto‐5′‐nucleotidase.British Journal of Pharmacology (2007) 152, 141–150; doi:10.1038/sj.bjp.0707361
α1‐Adrenoceptors and ejaculatory function
Authors: Michel M C 
Published: July 2007

The abnormal ejaculation of semen is a typical but infrequent side effect of someα1‐adrenoceptor antagonists, particularly those with selectivity forα1A‐adrenoceptors such as silodosin or tamsulosin. Recent clinical studies suggest that this represents a relative anejaculation rather than a retrograde ejaculation. An elegant study in this issue of the journal usingα1Asingle andα1A/B/Dtriple knock‐out mice reports a similar phenomenon in rodents. Using a multi‐disciplinary approach, the reduced ejaculation and related male infertility is shown to be caused by an impaired function of the vas deferens rather than by alterations in sperm formation, number or function. Similarities and differences between mouse and human data are discussed, particularly why a complete inhibition of all threeα1‐adrenoceptor subtypes has the strongest effects in mice whereas apparently onlyα1A‐adrenoceptor‐selective drugs impair ejaculatory function in humans.British Journal of Pharmacology(2007)152, 289–290; doi:10.1038/sj.bjp.0707369; published online 2 July 2007
Origin and evolution of high throughput screening
Authors: Pereira D A , Williams J A 
Published: July 2007

This article reviews the origin and evolution of high throughput screening (HTS) through the experience of an individual pharmaceutical company, revealing some of the mysteries of the early stages of drug discovery to the wider pharmacology audience. HTS in this company (Pfizer, Groton, USA) had its origin in natural products screening in 1986, by substituting fermentation broths with dimethyl sulphoxide solutions of synthetic compounds, using 96‐well plates and reduced assay volumes of 50‐100μl. A nominal 30mM source compound concentration provided high μM assay concentrations. Starting at 800 compounds each week, the process reached a steady state of 7200 compounds per week by 1989. Screening in the Applied Biotechnology and Screening Group was centralized with screens operating in lock‐step to maximize efficiency. Initial screens were full files run in triplicate. Autoradiography and image analysis were introduced for 125I receptor ligand screens. Reverse transcriptase (RT) coupled with quantitative PCR and multiplexing addressed several targets in a single assay. By 1992 HTS produced ‘hits’ as starting matter for approximately 40% of the Discovery portfolio. In 1995, the HTS methodology was expanded to include ADMET targets. ADME targets required each compound to be physically detected leading to the development of automated high throughput LC‐MS. In 1996, 90 compounds/week were screened in microsomal, protein binding and serum stability assays. Subsequently, the mutagenic Ames assay was adapted to a 96‐well plate liquid assay and novel algorithms permitted automated image analysis of the micronucleus assay. By 1999 ADME HTS was fully integrated into the discovery cycle.British Journal of Pharmacology (2007) 152, 53–61; doi:10.1038/sj.bjp.0707373
Telomerase and its potential for therapeutic intervention
Authors: Phatak P , Burger A M 
Published: July 2007

Telomerase and telomeres are attractive targets for anticancer therapy. This is supported by the fact that the majority of human cancers express the enzyme telomerase which is essential to maintain their telomere length and thus, to ensure indefinite cell proliferation – a hallmark of cancer. Tumours have relatively shorter telomeres compared to normal cell types, opening the possibility that human cancers may be considerably more susceptible to killing by agents that inhibit telomere replication than normal cells. Advances in the understanding of the regulation of telomerase activity and the telomere structure, as well as the identification of telomerase and telomere associated binding proteins have opened new avenues for therapeutic intervention. Here, we review telomere and telomerase biology and the various approaches which have been developed to inhibit the telomere/telomerase complex over the past decade. They include inhibitors of the enzyme catalytic subunit and RNA component, agents that target telomeres, telomerase vaccines and drugs targeting binding proteins. The emerging role of telomerase in cancer stem cells and the implications for cancer therapy are also discussed.British Journal of Pharmacology (2007) 152, 1003–1011; doi:10.1038/sj.bjp.0707374; published online 2 July 2007
Salt intake and insulin sensitivity in healthy human volunteers
Authors: Townsend Raymond R. , Kapoor Shiv , McFadden Christopher B. 
Published: July 2007

The literature on salt intake and insulin sensitivity presents a mixed picture, as some studies have shown an increase, whereas others have shown a decrease, in insulin action as sodium intake is enhanced. In some cases, this may relate to the study of salt intake in patients with co-morbidities such as hypertension or diabetes. In the present study, we selected healthy normotensive lean volunteers who underwent a euglycaemic clamp following 6 days of a low-salt diet (20 mmol sodium daily) and, subsequently, 6 days of a high-salt diet (200 mmol sodium daily). Our results show an increase in insulin-mediated glucose disposal during euglycaemic clamp conditions that was significantly higher following the high-salt diet compared with the low-salt diet (7.41±0.41 compared with 6.11±0.40 mg·kg−1 of body weight·min−1 respectively; P=0.03). We measured calf blood flow before and during insulin infusion (no significant change after the two dietary salt interventions was detected) and plasma non-esterified fatty acids (also no significant differences were detected). We observed the expected increases in renin concentration and aldosterone activity in subjects on the low-salt diet, and also observed a significantly less increase in plasma noradrenaline concentration during euglycaemic insulin infusion following the high-salt compared with the low-salt diet. We propose that the 4–5-fold increase in serum aldosterone and the greater increase in plasma noradrenaline concentration following the low-salt intervention compared with the high-salt period may have contributed to the differences in insulin sensitivity following the adjustment in dietary sodium intake.
Steps Used by the BLAST Algorithm: Figure 2.
Authors: Mount David W. 
Published: July 2007

INTRODUCTIONThe BLAST algorithm performs DNA and protein sequence similarity searches by an algorithm that is faster than FASTA but considered to be equally as sensitive. BLAST is very popular due to availability of the program on the World Wide Web through a large server at the National Center for Biotechnology Information (NCBI) and at many other sites. The BLAST algorithm has evolved to provide a set of very powerful search tools for the molecular biologist that are freely available to run on many computer platforms. This article provides a list of steps that describe how the BLAST algorithm searches a sequence database.
Cre Recombinase Gene Transfer In Vitro and Detection of loxP-Dependent Recombination: Figure 1.
Authors: Ohtsubo Kazuaki , Marth Jamey D. 
Published: July 2007

INTRODUCTIONAltering the genome of intact cells and organisms by site-specific DNA recombination has become an important gene-transfer methodology. The inclusion of exogenous recombinase target sequences within transferred DNA segments allows subsequent modifications to previously altered genomic structure that increase the utility of gene transfer and enhance experimental design. In this protocol, correctly targeted mouse embryonic stem (ES) cell clones bearing the F[tkneo] allele (containing several loxP sites) are subjected to in vitro Cre gene transfer to generate ES cell subclones bearing either Type 1 (Δ) or Type 2 (F) alleles. Type 2 ES cells are used to generate chimeric mice that are then crossed to germ-line Cre-expressing mice, such as ZP3-Cre transgenic mates. The additional time needed to breed the mice (~2-3 mo) is typically less troublesome than the cost and effort of maintaining multiple clone-derived lines of mice.
Imaging Retinotectal Synaptic Connectivity
Authors: Cohen-Cory Susana 
Published: July 2007

INTRODUCTIONThis protocol describes a method for imaging synaptic sites in individual axon or dendritic terminals in live, developing embryos. By selectively targeting expression of green fluorescent protein (GFP)-tagged pre- and post-synaptic proteins to developing neurons in vivo, it is possible to visualize synaptic sites and to correlate their distribution and dynamics with changes in the morphology of axon or dendritic arbors labeled with a red fluorescent molecule. The techniques described here may be applied to almost any neuronal circuit or neuron type, as long as the embryonic tissues are accessible for transfection and the neurons of interest are located at a depth that is within the working distance of the objective. These techniques have been optimized for Xenopus embryos, but they are applicable to other vertebrate systems such as zebrafish and chick embryos.
Maize Tissue Preparation and Extraction of RNA from Target Cells for Genotyping
Authors: Ohtsu Kazuhiro , Schnable Patrick S. 
Published: July 2007

INTRODUCTIONThe use of RNA for genotyping analysis can be advantageous because transcriptomes are significantly smaller than genomes and typically contain far fewer repetitive sequences. Laser capture microdissection (LCM) has been used successfully to isolate sequences (especially rare transcripts) that accumulate in specific tissues. Where the quantity of isolated material is limiting, amplification can be used to increase the amount of product. Upon conversion to cDNA, the product serves as template for 454 sequencing to produce expressed sequence tags for subsequent SNP analysis and detection.This protocol describes the preparation of acetone-fixed and paraffin-embedded maize seedling tissue sections. Once the sections are prepared, the PALM MicroBeam System, with its Laser Microdissection and Pressure Catapulting (LMPC) technology, is used to cut out cells of interest and “catapult” isolated tissues into collection caps. This tissue can then be used for RNA extraction.
SNP Mining from Maize 454 EST Sequences
Authors: Barbazuk W. Brad , Emrich Scott , Schnable Patrick S. 
Published: July 2007

INTRODUCTIONIn this protocol, 454 expressed sequence tags (ESTs) are generated by sequencing shoot apical meristem (SAM) cDNA from maize inbred lines on the 454 Life Sciences GS-20 sequencing system. The computational tool PolyBayes (Marth et al. 1999) is then used to identify single-nucleotide polymorphisms (SNPs). PolyBayes has been used successfully to identify SNPs in many different systems, including maize, and is particularly recommended for identifying SNPs in 454 sequences.
Single-Neuron Labeling Using the Genetic MARCM Method
Authors: Luo Liqun 
Published: July 2007

INTRODUCTIONOur brain is composed of hundreds of billions of neurons, each of which has an elaborate shape and a complex pattern of connections. To untangle this complexity, it is often useful to visualize one neuron at a time. This protocol describes single-neuron labeling using the MARCM system (Mosaic Analysis with a Repressible Cell Marker). The system was developed in Drosophila, but could, in principle, also work in other genetic model organisms such as Caenorhabditis elegans, zebrafish, or mice.
Construction of a Heated Incubation Chamber around a Microscope Stage for Time-Lapse Imaging: Figure 1.
Authors: Kulesa Paul M. , Kasemeier-Kulesa Jennifer C. 
Published: July 2007

INTRODUCTIONIntravital imaging of embryogenesis has the potential to provide valuable information on cell proliferation, cell shape changes, and cell migratory behaviors. However, most embryo model systems require a temperature-controlled environment. Several expensive commercially available temperature control devices have emerged, including microscope stages surrounded by custom-fit Plexiglas boxes, heated plates for culture dishes, and objective warmers for water-immersion lenses, that strictly control temperature and, in some cases, help control local gas mixtures. This protocol describes an easy-to-assemble, cost-effective, custom-made cardboard box and incubator, adaptable to each user’s specifications and microscope set-up. The cardboard box fits around the microscope, primarily the stage area, to assist in maintaining a prescribed temperature near the microscope stage. Warmed air, blown into the box enclosure from an incubator, circulates around the stage. The heated incubation box maintains a set temperature with minimal fluctuations and has been tested and utilized for studies of chick, mouse, and zebrafish embryogenesis.
Strategies for Sequence Similarity Database Searches
Authors: Mount David W. 
Published: July 2007

INTRODUCTIONDatabase similarity search programs tend to produce large volumes of output. It can become difficult to screen this volume of material and to assess whether the more remotely related sequences are really related to the query sequence. Thus, it is important to limit the sequence output; there are some relatively simple procedures that may be followed for each program, as described in this article. For searches of protein databases, avoid repetitive alignments with the same sequence by limiting searches to the protein sequence databases that are well curated, such as SwissProt and PIR, or to a specific genome, as opposed to the entire set of translated GenBank sequences (the GenPept database).
Using the Basic Local Alignment Search Tool (BLAST)
Authors: Mount David W. 
Published: July 2007

INTRODUCTIONThe BLAST algorithm was developed as a way to perform DNA and protein sequence similarity searches by an algorithm that is faster than FASTA but considered to be equally as sensitive. Both of these methods follow a heuristic (tried-and-true) method that almost always works to find related sequences in a database search, but does not have the underlying guarantee of an optimal solution like the dynamic programming algorithm. FASTA finds short common patterns in query and database sequences and joins these into an alignment. BLAST is similar to FASTA, but gains a further increase in speed by searching only for rarer, more significant patterns in nucleic acid and protein sequences. BLAST is very popular due to its availability on the World Wide Web through a large server at the National Center for Biotechnology Information (NCBI) and at many other sites. The BLAST algorithm has evolved to provide molecular biologists with a set of very powerful search tools that are freely available to run on many computer platforms. This article is intended to be a “user’s guide” to the principles underlying BLAST.
Changes of endocrine function of adipose tissue in anorexia nervosa: comparison of circulating levels versus subcutaneous mRNA expression
Authors: Dolezalova Radka , Lacinova Zdena , Dolinkova Marketa , Kleiblova Petra , Haluzikova Denisa , Housa Daniel , Papezova Hana , Haluzik Martin 
Published: July 2007

SummaryObjective  To study the influence of chronic malnutrition in patients with anorexia nervosa on endocrine function of adipose tissue on both circulating and subcutaneous fat mRNA expression level.Patients and design  A total of 12 patients with anorexia nervosa and 18 normal weight age‐matched women underwent anthropometric examination, single blood drawing and subcutaneous adipose tissue biopsy.Measurements  Serum concentrations of high‐sensitive CRP (hsCRP), leptin, soluble leptin receptor, adiponectin, resistin, interleukin‐6 and insulin were measured by Luminex, enzyme‐linked immunosorbent assay (ELISA) and radioimmunoassay (RIA) kits. Subcutaneous adipose tissue mRNA expression of the same adipokines, adiponectin receptors 1 and 2 and immunocompetent cells marker CD68 was measured by real‐time polymerase chain reaction (PCR).Results  Decreased body fat content of patients with anorexia nervosa was accompanied by reduced hsCRP, leptin and increased adiponectin and soluble leptin receptor. Resistin, interleukin‐6 and insulin levels did not differ from those of the control group. Fat mRNA adiponectin, leptin, interleukin‐6 and CD68 expression was reduced, resistin mRNA expression was increased and adiponectin receptor 1 and 2 expression were unchanged as compared to the control group.Conclusions  Local perturbations in resistin, adiponectin and interleukin‐6 mRNA expression in subcutaneous adipose tissue are not reflected by its circulating levels. These changes could be involved in some local metabolic disturbances in subcutaneous adipose tissue of anorexia nervosa patients.