bluuebunny/crossref_metadata_2025_split huggingface dataset:
Non-alcoholic fatty liver disease: an overview of prevalence, diagnosis, pathogenesis and treatment considerations
Authors: Preiss David , Sattar Naveed 
Published: August 2008

The global increase in the prevalence of obesity has heralded a rise in associated liver injury namely NAFLD (non-alcoholic fatty liver disease). It is estimated that 20–30% of adult populations in developed countries have NAFLD and, although high quality data is currently lacking, the condition is clearly increasing in children also. NAFLD should be suspected in those with commonly available simple clinical signs and biochemistry consistent with insulin resistance. A small number of individuals with NAFLD, often considered a relatively benign condition, will progress to more severe stages of liver disease including NASH (non-alcoholic steatohepatitis) with or without fibrosis, cirrhosis and occasionally hepatocellular carcinoma. NAFLD is also commonly associated with an increased risk of developing Type 2 diabetes and treatable features of insulin resistance such as dyslipidaemia and dysglycaemia. Histological examination of liver tissue remains the only proven method to distinguish between simple steatosis and NASH, a condition far more likely to progress to cirrhosis. Identification of an imaging technique or non-invasive marker to achieve this distinction is therefore much sought after and would allow larger clinical trials and better clinical assessment. Case series and pilot studies of lifestyle advice, insulin sensitizers and other medications have shown improvements in liver histology and serum liver enzymes but robust randomized controlled studies are needed. Furthermore, the cost/benefit ratio of any new therapies, and any potential harms, must be evaluated carefully before being clinically advocated.
In Search of the Active Ingredient: What Really Works in Mental Health Care?
Authors: Brown G.S. (Jeb) , Cameron Joanne , Brown Lindsey 
Published: August 2008

AbstractThis brief report provides a concise overview of decades of research into what really makes a difference in treatment outcomes for the most common psychiatrics disorders. The evidence points to the conclusion that the relationship with the clinician accounts for much more of the variance in treatment outcomes than the treatment methods themselves. The discussion of research and statistical methodology are relevant to investigations of treatments for speech and language disorders.
Treating the Gifted Client
Authors: Williams Dale F. 
Published: August 2008

Abstract
Treating gifted individuals can be both rewarding and challenging. As clients, they often present a thorough understanding of the therapy plan and offer worthwhile perspectives. However, this population’s tendency toward perfectionism can make it difficult to get them out of their comfort zones, a necessary part of stuttering therapy. Procedures such as modeling, setting attainable goals, and accelerating the instruction can help them experience success and transfer skills to environments outside the clinic room.
Auditory Discrimination Deficits in Boys Who Stutter: A Preliminary Investigation
Authors: LaSalle Lisa R. , Duginske Rachel D. 
Published: August 2008

Abstract
Our purpose was to examine the basic level of auditory processing skills in school-age children who stutter. We administered three auditory processing tasks—dichotic digits, temporal patterning, and auditory discrimination—from the Differential Screening Test for Processing (Richard & Ferre, 2006). Five 6- to 9-year-old boys who stutter were age-matched to five boys who do not stutter. Children who stutter showed significantly poorer performance on the auditory discrimination subtest. These preliminary results suggest that, for a subgroup of children who stutter, auditory discrimination difficulties may exacerbate stuttering and thus be a factor worth considering in clinical intervention.
The PAS Reaction for Staining Cell Walls
Authors: Baum Stuart 
Published: August 2008

INTRODUCTIONThe periodic acid Schiff (PAS) reaction is useful for looking at cell walls, especially in meristematic tissues. Total carbohydrates of insoluble polysaccharides (e.g., starch and cellulose) stain magenta to purplish red. Toluidine blue O or fast green is routinely used as a counterstain to improve contrast. In the gynoecium of the Arabidopsis flower, the transmitting tract cells are made of acid proteoglycans and acid glycoproteins. The cell walls of these cells should be stained with Alcian Blue instead of using the PAS reaction. Alcian Blue, which stains the walls a dark blue, can be used alone or as a counterstain with the PAS reaction to highlight cell walls that do not react with Alcian Blue.
Analyzing DNA Replication III: Antibody Labeling of Incorporated Bromodeoxyuridine (BrdU) in Tissues and Cells: Figure 1.
Authors: Jackson Dean , Cook Peter R. 
Published: August 2008

INTRODUCTIONThe number of cells traversing the cell cycle and the rate of progression through it provide important indices of cell growth and tumorigenicity. S-phase cells can also be identified by their high content of DNA polymerase and proliferating cell nuclear antigen, a component of the leading-strand polymerase. Although both these markers can be detected rapidly and conveniently using the appropriate antibodies, neither are found exclusively in S-phase cells. Immunolabeling after incorporation of modified DNA precursors (e.g., 5-bromodeoxyuridine [BrdU, bromodeoxyuridine]) allows more rapid and precise detection of cells in S-phase of the cell cycle. BrdU is phosphorylated by cells to give BrdUTP, and this precursor is incorporated into DNA instead of deoxythimidine triphosphate. In living cells, BrdU is incorporated into replication sites that can then be detected using fluorochrome or enzyme-coupled antibodies. Alternatively, DNA synthesis sites can be labeled at high resolution by incubating cells with analogs of the natural precursors of DNA. After fixation to preserve nuclear morphology, the DNA duplex is denatured to allow antibodies access to the BrdU. Cells labeled in this way either in vivo or in vitro display a few hundred discrete nuclear sites early in S-phase, with distinct patterns of DNA replication that are characteristic of different stages of S-phase. This protocol describes two commonly used methods of denaturation, as well as techniques for antibody labeling of mounted tissues and encapsulated cells.
In Vitro Assays for the Extracellular Matrix Protein-Regulated Extravasation Process
Authors: Ma Chaoyu , Wang Xiao-Fan 
Published: August 2008

INTRODUCTIONExtravasation, the process by which circulating tumor cells pass through the blood vessel wall, is a critical step of metastasis. Extracellular matrix (ECM) proteins secreted by the cancer cells are likely to play an interactive role in the dynamic interaction between cancer cells and endothelial cells during the extravasation process. This protocol describes two in vitro assays, the transendothelial cell migration (TEM) assay and the vascular permeability assay, which are used to demonstrate the involvement of ECM proteins in cancer cell extravasation. Both assays employ primary human umbilical vein endothelial cells (HUVEC) to reconstitute a vessel wall (HUVEC monolayer) on a porous filter membrane within a Transwell chamber. The TEM assay examines the efficiency of cancer cells to migrate through the vessel by co-culturing the cancer cells with the endothelial monolayer. The vascular permeability assay allows the study of the impact of secreted ECM proteins on the permeability of the vessel wall by applying conditioned medium from cancer cells to the endothelial monolayer.
Whole Mount RNA In Situ Hybridization on Zebrafish Embryos: Mounting
Authors: Moens Cecilia 
Published: August 2008

INTRODUCTIONThis protocol describes the mounting of zebrafish embryos for observation after whole-mount in situ hybridization. Embryos can be mounted in glycerol or dehydrated and mounted in Permount, with or without the yolk. Glycerol causes less shrinkage, but is not as effective a clearing agent. Embryos mounted on the yolk should be stored in the dark and photographed immediately, because the yolks darken rapidly.
Isolation of Centrosomes from Cultured Mammalian Cells
Authors: Meigs Thomas E. , Kaplan Daniel D. 
Published: August 2008

INTRODUCTIONThe centrosome is a cell’s primary microtubule-organizing center. In most mammalian cells, the centrosome is composed of a pair of centrioles and surrounding pericentriolar material. The centrosome is duplicated exactly once per cell cycle such that at the onset of mitosis, a cell has two centrosomes, which serve as poles of the mitotic spindle. During cytokinesis, one centrosome is segregated to each daughter cell. This protocol describes the isolation of centrosomes from asynchronous cells, and thus the purified material will consist primarily of interphase centrosomes. Isolated centrosomes can be used in a variety of assays, including studies of microtubule function and the identification of centrosome-associated proteins and their interactions.
Imaging Calcium Dynamics Using Targeted Recombinant Aequorins
Authors: Pozzan Tullio , Rizzuto Rosario 
Published: August 2008

INTRODUCTIONAequorin is a small protein produced by the genus Aequorea that was widely used in the 1960s and 1970s as a probe to measure Ca2+ in living cells. The invention of the carboxylate Ca2+ indicators, which are much simpler to load into intact living cells and to calibrate and image at the single-cell level, has led most groups to abandon aequorin. Yet, this latter Ca2+ indicator still offers some advantages over the fluorescent probes. In particular, the use of molecular biological techniques for expressing recombinant aequorin in mammalian cells, thus eliminating the need for microinjection, has opened new possibilities for this probe. Among the new uses of aequorin, one of the most interesting is the potential for targeting it specifically to different cellular locations, thus opening the possibility of monitoring selectively the dynamics of [Ca2+] with unprecedented spatial resolution. This article briefly discusses the problems concerned with targeting aequorin to different locations, the advantages and disadvantages offered by the steep dependence of luminescence on [Ca2+], and the instruments needed to obtain reliable measurements.
Genetic analysis of the Hispano‐Breton heavy horse
Authors: Pérez‐Gutiérrez L. M. , De la Peña A. , Arana P. 
Published: August 2008

SummaryHispano‐Breton (HB) is a horse breed with a recent mixed ancestry. It was developed in the 1930s by crossing local mares with Breton draught horses imported from France. Nowadays it is considered to be in a vulnerable situation due to census decline. To genetically characterize the breed and to set up the basis for a conservation programme, we have employed two types of molecular markers: a 347‐bp D‐loop mitochondrial DNA (mtDNA) fragment and 13 microsatellite loci. A representative sample of 53 HB individuals was analysed together with a sample of 40 Pura Raza Española horses for comparison. Both types of markers revealed a high level of genetic diversity in the HB breed, emphasizing the importance of its conservation. The construction of a phylogenetic network with mtDNA sequences including various Iberian breeds and European heavy horses provided an overall picture of the ubiquitous appearance of HB matrilines with respect to other breeds and revealed the singularity of certain HB maternal lineages. Despite the high allelic richness found in HB horses, microsatellite analysis evidenced a certain degree of inbreeding as a consequence of the type of management generally used for local breeds.
Genetic variation in the mannosidase 2B2 gene and its association with ovulation rate in pigs
Authors: Campbell E. M. , Nonneman D. J. , Kuehn L. A. , Rohrer G. A. 
Published: August 2008

SummaryOvulation rate is an important phenotypic trait that is a critical component of litter size in pigs. Despite being moderately heritable in pigs, selection for increased ovulation rate is difficult because it is difficult to measure and is a sex‐limited trait. A QTL for ovulation rate residing on the p‐terminal end of pig chromosome 8 has been detected in a Meishan‐cross resource population. Comparative analysis of this region yielded a positional candidate gene, mannosidase 2B2 (MAN2B2), for this QTL. The entire coding region of MAN2B2 was resequenced in the Meishan and White Composite founder animals of the resource population to identify SNPs. Eleven polymorphisms that alter the protein product of MAN2B2 were discovered and tested for statistical associations with ovulation rate in three generations of the resource population. The polymorphism located at position 1574 of the mRNA (D28521:c.1574A>G) was the most significant polymorphism tested (P = 0.00005) where the additive effect of the c.1574A allele was estimated to be −0.89 ova. This polymorphism was determined to be more significantly associated with ovulation rate than the breed‐specific analysis conducted during the line‐cross QTL discovery. The c.1574A>G marker was not associated with ovulation rate in an occidental population. Therefore, either MAN2B2 has a unique epistatic interaction within the Meishan‐cross population or the c.1574A>G SNP is in linkage disequilibrium with the actual causative genetic variation in the Meishan‐cross population.
Differences and similarities in the developmental status of embryo‐derived stem cells and primordial germ cells revealed by global expression profiling
Authors: Mise Nathan , Fuchikami Takuya , Sugimoto Michihiko , Kobayakawa Satoru , Ike Fumio , Ogawa Takehiko , Tada Takashi , Kanaya Shigehiko , Noce Toshiaki , Abe Kuniya 
Published: August 2008

Embryonic germ‐line cells are unipotent cells that give rise to either sperm or oocytes. However, pluripotent stem cells can be derived from primordial germ cells (PGCs) or spermatogonia, suggesting that germ‐line cells retain a capacity for pluripotency. Here, we made genome‐wide comparisons of the gene expression profiles of freshly isolated PGCs, in vitro‐formed PGCs (iPGCs), and other stem cell lines, including embryonic stem cells (ESCs), embryonic germ cells (EGCs) and germ‐line stem (GS) cells. Comparing PGC with ESC, 382 genes/transcripts were significantly up‐regulated in ESC, while 188 were elevated in PGC. This suggests that PGCs possess transcription program distinct from that of ESC, although both share expression of many pluripotency‐associated genes.Our micro‐array analysis showed that the analyzed samples could be classified into two groups: one consisting of all the ESCs and most of EGCs, and the other containing PGC samples, iPGC, one type of female EGC and GS cells. We then identified “signature” genes for the two groups, and used them to characterize GS cells, EGC, and iPGCs, and revealed developmental status of each cell type. The relationships between PGCs and stem cells derived from embryos or germ cells are discussed in light of these findings.
p97 homologs from Caenorhabditis elegans, CDC‐48.1 and CDC‐48.2, suppress the aggregate formation of huntingtin exon1 containing expanded polyQ repeat
Authors: Nishikori Shingo , Yamanaka Kunitoshi , Sakurai Toshihiko , Esaki Masatoshi , Ogura Teru 
Published: August 2008

Polyglutamine (polyQ)‐expanded proteins are associated with cytotoxicity in some neurodegenerative disorders such as Huntington's disease. We have reported that the aggregation of the polyQ‐expanded protein is partially suppressed by co‐expression of either of two homologs of an AAA chaperone p97, CDC‐48.1 or CDC‐48.2, in Caenorhabditis elegans, but how p97 regulates the aggregation of polyQ‐expanded proteins remains unclear. Here we present direct evidence that CDC‐48.1 and CDC‐48.2 suppress the aggregation of a huntingtin (Htt) exon1 fragment containing an expanded polyQ repeat in vitro. CDC‐48.1 and CDC‐48.2 bound the Htt exon1 fragment directly, and suppressed the formation of SDS‐insoluble aggregates of Htt fragments containing 53 glutamine residues (HttQ53) independently of nucleotides. CDC‐48.1 and CDC‐48.2 also modulated the oligomeric states of HttQ53 during the aggregate formation. In the absence of CDC‐48.1 and CDC‐48.2, HttQ53 formed 70–150 kDa oligomers, whereas 300–500 kDa oligomers as well as 70–150 kDa oligomers accumulated in the presence of CDC‐48.1 and CDC‐48.2. Taken together, these results suggest that p97 plays a protective role in neurodegenerative disorders by directly suppressing the protein aggregation as a molecular chaperone.
Nurses’ knowledge of pharmacology behind drugs they commonly administer
Authors: Ndosi Mwidimi E , Newell Rob 
Published: August 2008

Aim.  To determine if nurses had adequate pharmacology knowledge of the drugs they commonly administer.Background.  Literature suggests that nurses have insufficient pharmacology knowledge. We also know that nurses and teachers of pharmacology are not satisfied with the amount of pharmacology taught in preregistration programmes in the UK. There is a lack of primary research on nurses’ knowledge of pharmacology for the purpose of drug administration.Design.  We used a non‐experimental causal comparative and correlational design.Methods.  We recruited a convenience sample of 42 nurses working in surgical wards of a foundation hospital in the North of England. Data were collected by structured interview and questionnaire methods. During the interview, the participants made a blinded selection of one out of four drugs they commonly administer and answered standard questions which focused on specific pharmacology knowledge. Their answers were given a score out of 10 (100%) to determine their actual pharmacology knowledge.Results.  The sample comprised of 18 (42·9%) junior nurses and 24 (57·1%) senior nurses. They had a median experience of 10·87 years postregistration. Their mean knowledge score was six ranging between two and nine (SD 1·9). Only 11 (26·1%) nurses scored eight or above and the majority 24 (57·2%) scored below seven, indicating inadequate knowledge. Knowledge of the mechanism of action and drug interactions was poor. There was a correlation between knowledge and experience.Conclusions.  The results of this study suggest that nurses have inadequate knowledge of pharmacology. The results will contribute to the evidence of nurses’ knowledge of pharmacology in the UK.Relevance to clinical practice.  This study supports the need for supplementary pharmacology education for nurses in clinical settings, focusing on common drugs they administer. This will increase nurses’ knowledge and confidence in drug administration and safer medicines management.
The vacuolar transport of aleurain‐GFP and 2S albumin‐GFP fusions is mediated by the same pre‐vacuolar compartments in tobacco BY‐2 and Arabidopsis suspension cultured cells
Authors: Miao Yansong , Li Kwun Yee , Li Hong‐Ye , Yao Xiaoqiang , Jiang Liwen 
Published: August 2008

SummarySoluble proteins reach vacuoles because they contain vacuolar sorting determinants (VSDs) that are recognized by vacuolar sorting receptor (VSR) proteins. Pre‐vacuolar compartments (PVCs), defined by VSRs and GFP‐VSR reporters in tobacco BY‐2 cells, are membrane‐bound intermediate organelles that mediate protein traffic from the Golgi apparatus to the vacuole in plant cells. Multiple pathways have been demonstrated to be responsible for vacuolar transport of lytic enzymes and storage proteins to the lytic vacuole (LV) and the protein storage vacuole (PSV), respectively. However, the nature of PVCs for LV and PSV pathways remains unclear. Here, we used two fluorescent reporters, aleurain‐GFP and 2S albumin‐GFP, that represent traffic of lytic enzymes and storage proteins to LV and PSV, respectively, to study the PVC‐mediated transport pathways via transient expression in suspension cultured cells. We demonstrated that the vacuolar transport of aleurain‐GFP and 2S albumin‐GFP was mediated by the same PVC populations in both tobacco BY‐2 and Arabidopsis suspension cultured cells. These PVCs were defined by the seven GFP‐AtVSR reporters. In wortmannin‐treated cells, the vacuolated PVCs contained the mRFP‐AtVSR reporter in their limiting membranes, whereas the soluble aleurain‐GFP or 2S albumin‐GFP remained in the lumen of the PVCs, indicating a possible in vivo relationship between receptor and cargo within PVCs.
Coronatine Alleviates Polyethylene Glycol‐induced Water Stress in Two Rice (Oryza sativa L.) Cultivars
Authors: Ai L. , Li Z. H. , Xie Z. X. , Tian X. L. , Eneji A. E. , Duan L. S. 
Published: August 2008

AbstractWe studied the effects of coronatine (COR), a structural and functional analogue of jasmonates, on the drought tolerance of two rice cultivars, Handao 297 (a drought‐tolerant upland rice) and Yuefu (a drought‐sensitive lowland rice). Seedlings were treated with COR at the three‐leaved stage at 0.01 and 0.1 μm for 24 h, followed by imposition of water deficit induced with 20 % polyethylene glycol (PEG). Water stress reduced the biomass of both cultivars and increased leaf lipid peroxidation and solute leakage. Pre‐treatment with COR significantly increased the activities of superoxide dismutase, catalase, ascorbate peroxidase and glutathione reductase in leaf tissues of water‐stressed Handao 297 (0.01 μm) and Yuefu (0.1 μm) seedlings. COR supplement also increased the accumulation of 44 and 32 kDa polypeptides in water‐stressed Handao 297 (0.01 and 0.1 μm) and Yuefu (0.1 μm) and significantly induced the expression of 29 kDa polypeptide in Handao 297. The results suggest that COR might alleviate drought stress by activating antioxidant enzymes and inducing proteins, thereby preventing membrane peroxidation and denaturation of bio‐molecules. Thus, membrane permeability decreased substantially by 24–27 % in Handao 297, and 22–29 % in Yuefu. The optimal concentrations conferring drought resistance were 0.01 for the upland rice and 0.1 μm for the lowland cultivar.
Annual dynamics of the mesozooplankton communities in a highly variable ecosystem (North Adriatic Sea, Italy)
Authors: Camatti Elisa , Comaschi Alessandra , De Olazabal Alessandra , Fonda Umani Serena 
Published: August 2008

AbstractMesozooplankton community composition at the species/taxa level was investigated from January to December 2001, at eight stations in the northern Adriatic Sea. Annual dynamics, taxonomic composition and spatial diversity in relation to different trophic conditions are discussed as related to previous studies and to different conditions, such as the presence of mucilage events. Zooplankton communities all over the northern sub‐basin were dominated by the cladoceran Penilia avirostris in summer, and by the calanoids Paracalanus parvus, Acartia clausi and the poecilostomatoids Oncaea spp. during the rest of the year. Whereas coastal communities were more variable with time and location, we were able to identify a group of offshore stations with a similar species/taxa composition and annual dynamics. Comparing the present results with previous studies, we observed significant changes in community composition in the time scale of 20 years, mostly due to a general decrease of A. clausi as the dominant species, being replaced by P. parvus. Also, P. avirostris swarms appear to have extended their temporal occurrence, and were present for longer periods of time when compared to past records. These changes might be related to the observed general increase of the average sea water temperature in the northern Adriatic Sea.