bluuebunny/crossref_metadata_2025 huggingface dataset:
Review: Antiplatelet Drugs: What Comes Next?
Authors: Kei Anastazia A. , Florentin Matilda , Mikhailidis Dimitri P. , Elisaf Moses S. , Liberopoulos Evangelos N.
Published: November 2010

Antiplatelet drugs are important components in the management of atherothrombotic vascular disease. However, several limitations restrict the safety and efficacy of current antiplatelet therapy in clinical practice. Interpatient variability and resistance to aspirin and/or clopidogrel has spurred efforts for the development of novel agents. Indeed, several antiplatelet drugs are at various stages of evaluation; those at advanced stage of development are the focus of this review.
Exogenousl-Arginine and HDL Can Alter LDL and ox-LDL-Mediated Platelet Activation
Authors: Sener Azize , Enc Elif , Ozsavci Derya , Vanizor-Kural Birgul , Yanikkaya-Demirel Gulderen , Oba Rabia , Uras Fikriye , Demir Muzaffer
Published: November 2010

The aim of this study is to investigate the effects of exogenous l-arginine and HDL on LDL and oxidized LDL (ox-LDL)-mediated platelet activation. Adenosine diphosphate (ADP)-activated platelets have been incubated with lipoproteins with or without l-arginine. P-selectin receptor numbers per platelet have been measured by flow cytometry. After incubation with only l-arginine (without lipoproteins), platelet nitric oxide (NO) levels and P-selectin receptor numbers significantly increased compared to the controls ( P < .05). After incubation with LDL or ox-LDL, receptor numbers of P-selectin significantly increased ( P < .001). However, P-selectin receptor numbers in platelets treated with l-arginine + LDL or l-arginine + ox-LDL decreased compared to the levels in platelets treated with only LDL or ox-LDL ( P < .01, P < .001, respectively). Addition of HDL to l-arginine + ox-LDL caused significant reduction in P-selectin receptor numbers as in the control values ( P < .001).We have concluded that l-arginine causes enhanced platelet NO levels and blocks the effects of LDL or ox-LDL on platelet P-selectin receptor numbers and HDL also strengthens this effect of l-arginine.
Genetic Mutations in Turkish Population With Pulmonary Embolism and Deep Venous Thrombosis
Authors: Kupeli Elif , Verdi Hasibe , Simsek Abdullah , Atac Fatma Belgin , Eyuboglu Fusun Oner
Published: November 2010

Venous thromboembolism (VTE) is a universal health hazard. Inherited and acquired risk factors increase the risk of VTE. We evaluated the relationship between factor V (G1691A, A1090G, and A1299G), prothrombin (PT G20210A), methylenetetrahydrofolate reductase (MTHFR C677T) mutations, plasminogen activator inhibitor 1 (PAI-1 -675) polymorphism, and VTE in Turkish population. In all, 80 patients with VTE and 104 controls were included. Heterozygous factor V Leiden (FVL) mutation was significantly higher among patients ( P = .04) with allele frequency of 6.3% ( P = .01). Heterozygous PT G20210A mutation was also significantly higher among patients ( P = .001) with allele frequency of 6.9% ( P = .003). MTHFR 677TT genotype was significantly higher in patients ( P = .009) with allele frequency of 23.8% ( P = .005). No significant difference was found in FV A1090G and FV A1299G mutation rate as well as PAI-1 genotypes and their allele frequencies ( P > .05). Thus, frequencies of FV G1691A, PT G20210A, and MTHFR C677T mutations are higher in patients with VTE. FV A1090G, FV A1299G mutations, and PAI-1 gene polymorphisms may not be a risk factor for VTE in Turkish population.
Safety and Efficacy of Thrombin-JMI: A Multidisciplinary Expert Group Consensus
Authors: Bhandari Mohit , Ofosu Frederick A. , Mackman Nigel , Jackson Craig , Doria Cataldo , Humphries John E. , Babu Sateesh C. , Ortel Thomas L. , Hoffman Van Thiel David , Walenga Jeanine M. , Wahi Rakesh , Teoh Kevin Hong To , Fareed Jawed
Published: November 2010

Background: The use of bovine thrombin has been an effective approach to aiding hemostasis during surgery for over 60 years. Its use has a reported association with the development of antibodies to coagulation factors with limited evidence to the clinical significance. Methods: The Collaborative Delphi survey methodology was used to develop a consensus on specified topic areas from a panel of 12 surgeons/scientists who have had experience with topical thrombins; it consisted of 2 rounds of a Web-based survey and a final live discussion. Results: Some key issues that reached consensus included: bovine, human plasma-derived and recombinant human thrombin are equally effective hemostatic agents with similar adverse event rates, and immunogenicity to a topical protein rarely translate into adverse events. Conclusions: Although a risk of immunogenicity is associated with all topical thrombins, no conclusive clinical evidence is available that these antibodies have any significant effect on short- and long-term clinical consequences.
Commitment and Volunteer-Related Outcomes Among Private Club Board and Committee Member Volunteer Leaders
Authors: Cha JaeMin , Cichy Ronald F. , Kim SeungHyun
Published: November 2010

This study examined the direct and indirect factors influencing volunteer leaders’ future intentions to continue to serve as board and committee members in private clubs. The proposed model was tested with a sample of 457 board and committee members, using mainly a Web survey. Findings showed that the volunteer leaders’ affective commitment had a direct and positive influence on both their volunteer dedication and involvement, whereas their normative commitment had a direct and positive influence only on their volunteer dedication. Both volunteer dedication and involvement were found to be important, direct factors for volunteer leaders’ future volunteer intentions to continue to serve as board and committee members in private clubs.
An Integrated Model of Employees’ Behavioral Intention Toward Innovative Information and Communication Technologies in Travel Agencies
Authors: Cheng Simone , Cho Vincent
Published: November 2010

This study proposes a comprehensive model that incorporates attitudinal variables in the technology acceptance model, motivational variables in the innovation diffusion theory, and the social variable in the theory of planned behavior to investigate the attitude, behavioral intentions, and usage of information and communication technologies (ICT) by employees in Hong Kong travel agencies. It complements trends in current research on technology adoption in the tourism industry. A survey was conducted with 190 employees from travel agencies, and structural equation modeling was used to analyze the data. Our results show that the integrated model of ICT adoption is adequate to predict employees’ adoption behavior. Furthermore, the perceived usefulness, perceived ease of use, trialability, and observability of new ICT are the major factors stimulating employees’ attitude toward it. Based on our findings, both theoretical and practical suggestions for stimulating employees’ adoption of ICT are proposed.
Does the Market Care About RevPAR? A Case Study of Five Large U.S. Lodging Chains
Authors: Chen Jianan , Koh Yoon , Lee Seoki
Published: November 2010

During the past several decades, the lodging industry has used RevPAR (revenue per available room) as a key indicator to evaluate a firm’s performance and to make investment decisions. However, a limited number of research articles empirically examined whether or not RevPAR, in fact, is a valid measure for a lodging firm’s performance, especially when compared with other traditional performance measures. The purpose of the current discussion, therefore, is to compare the explanatory power of RevPAR with three traditional financial measures—earnings per share (EPS), return on assets (ROA), and return on equity (ROE)—for lodging firms’ performance, estimated by total shareholders’ returns.
The diagnosis and management of common non-specific back pain - a clinical review
Authors: Brown Grahame
Published: November 2010

The assessment and management of common non-specific back pain that is associated with considerable personal morbidity and cost to society is reviewed and set in a bio-psycho-social context.
Feasibility of an Intensive Trial of Modern Dance for Adults with Parkinson Disease
Authors: Batson Glenna
Published: November 2010

Although preliminary evidence supports the psychophysical benefits of dance for adults with Parkinson disease, guidelines for community-based dance programs await further scrutiny for safety and efficacy. This pilot study was designed to assess the feasibility of an intensive trial of group-delivered modern dance for 11 adults with early-to-middle stage Parkinson’s. The Timed ‘‘Up and Go’’ test and the Fullerton Advanced Balance scale were administered to assess balance safety and re-administered at closure along with a self-reported feedback questionnaire. Video recordings were analyzed for qualitative behavioral change. Pre/posttest comparisons from the Timed ‘‘Up and Go’’ test were not significant, while those from the Fullerton Advanced Balance Scale were significant at the .05 level for the group as a whole (p = .01) with an average score change of +3.1 points. Although qualitative results generally concurred with the social benefits associated with an enjoyable form of expressive exercise, important indicators surfaced bearing on future research and community program designs. These include need for more rigorous stratification of participants and adapting dance class structure to address specific group needs to promote motor learning for sustained functional gains.
The Psychology of Qi Gong: A Qualitative Study
Authors: Posadzki Paul
Published: November 2010

This article presents an in-depth, qualitative study, analyzing responses to focus-group interviews regarding individuals’ self-reported experiences and attitudes during Qi Gong practice. Semistructured interviews were conducted with three Qi Gong groups in order to collect research data. These data were transcribed verbatim and subjected to content and thematic analysis across and within groups. The analysis indicates extraordinary experiences of Qi Gong practitioners on various levels of bio-psycho-spiritual/energetic functioning. The results indicate how Qi Gong influences the complexity and multidimensionality of individuals’ health. In the discussion, the author compares and contrasts his results with other recently performed research.
The association of bile acid excretion and atherosclerotic coronary artery disease
Authors: Charach Gideon , Grosskopf Itamar , Rabinovich Alexander , Shochat Michael , Weintraub Moshe , Rabinovich Pavel
Published: November 2010

Background: Excess cholesterol is usually eliminated from the body by conversion to bile acids excreted in feces as bile salts. The excretion of large amounts of bile protects against atherosclerosis, while diminished excretion may lead to coronary artery disease (CAD).

Objective: To investigate a relationship between CAD and bile acid excretion.

Methods: Bile acid excretion was compared between 36 patients with proven CAD and 37 CAD-free individuals (controls). The groups were comparable for demographics and selected risk factors. All subjects received a 4-day standard diet that included ∼500 mg of cholesterol. Fecal bile acids from 24-hour stool collections were measured by gas liquid chromatography.

Results: CAD patients excreted lower amounts of total bile acids (358 ± 156 mg) than controls (617 ± 293 mg; p < 0.01) and less deoxycholic acid (188.29 ± 98.12 mg versus 325.96 ± 198.57 mg; p < 0.0001) and less lithocholic acid (115.43 ± 71.89 mg versus 197.27 ± 126.87 mg; p < 0.01). Advanced age, male gender, left ventricular ejection fraction and total bile acid levels were significant independent factors that predicted CAD ( p < 0.05). Mortality, CAD and cerebrovascular accident development rates were significantly lower for the controls at the 13-year follow up.

Conclusion: CAD patients have significantly decreased bile acid excretion levels than non-CAD patients. An impaired ability to excrete cholesterol may be an additional risk factor for CAD development.
Indoleamine 2,3-dioxygenase specific, cytotoxic T cells as immune regulators
Authors: Sørensen Rikke Bæk , Hadrup Sine Reker , Svane Inge Marie , Hjortsø Mads Christian , thor Straten Per , Andersen Mads Hald
Published: November 2010

Indoleamine 2,3-dioxygenase (IDO) is an immunoregulatory enzyme that is implicated in suppressing T-cell immunity in normal and pathologic settings. Here, we describe that spontaneous cytotoxic T-cell reactivity against IDO exists not only in patients with cancer but also in healthy persons. We show that the presence of such IDO-specific CD8+ T cells boosted T-cell immunity against viral or tumor-associated antigens by eliminating IDO+ suppressive cells. This had profound effects on the balance between interleukin-17 (IL-17)–producing CD4+ T cells and regulatory T cells. Furthermore, this caused an increase in the production of the proinflammatory cytokines IL-6 and tumor necrosis factor-α while decreasing the IL-10 production. Finally, the addition of IDO-inducing agents (ie, the TLR9 ligand cytosine-phosphate-guanosine, soluble cytotoxic T lymphocyte–associated antigen 4, or interferon γ) induced IDO-specific T cells among peripheral blood mononuclear cells from patients with cancer as well as healthy donors. In the clinical setting, IDO may serve as an important and widely applicable target for immunotherapeutic strategies in which IDO plays a significant regulatory role. We describe for the first time effector T cells with a general regulatory function that may play a vital role for the mounting or maintaining of an effective adaptive immune response. We suggest terming such effector T cells “supporter T cells.”
Irf8 regulates macrophage versus neutrophil fate during zebrafish primitive myelopoiesis
Authors: Li Li , Jin Hao , Xu Jin , Shi Yuqian , Wen Zilong
Published: November 2010

In vertebrates, myeloid cells comprise polymorphonuclear and mononuclear lineages that arise from 2 successive waves of development: a transitory primitive wave giving rise to limited myeloid cells during embryonic stage and a definitive wave capable of producing myeloid cells throughout the fetal and adult life. One key unresolved question is what factors dictate polymorphonuclear versus mononuclear lineage fates during myelopoiesis. Here we show that during zebrafish embryogenesis interferon regulatory factor-8 (irf8) is expressed specifically in macrophages but not neutrophils. Suppression of Irf8 function in zebrafish causes a depletion of macrophages and an enhanced output of neutrophils but does not affect the overall number, proliferation, and survival of primitive myeloid cells. These data indicate that the skewed myeloid lineage development in Irf8 knockdown embryos results from a cell-fate switching. Such a conclusion is further supported by the observation showing that overexpression of Irf8 promotes macrophage formation at the expense of neutrophil development. Genetic epistasis analysis reveals that Irf8 acts downstream of Pu.1 but is insufficient to promote macrophage development in the absence of Pu.1. Our findings demonstrate that Irf8 is a critical determinant for neutrophil versus macrophage fate choice during zebrafish primitive myelopoiesis.
Hematogones: a new prognostic factor for acute myeloblastic leukemia
Authors: Chantepie Sylvain P. , Salaün Véronique , Parienti Jean-Jacques , Truquet Florence , Macro Margaret , Cheze Stephane , Vilque Jean Pierre , Reman Oumedaly
Published: November 2010

Acute myeloid leukemia (AML) patient outcomes remain heterogeneous, and new prognostic tools are needed to assess the risk of relapse. Hematogones (HGs) are normal B-lymphocyte precursors, which increase in number in hematologic diseases. The prognostic impact of the presence of detectable HGs on the leukemia-free survival (LFS) and overall survival of 120 consecutive patients with AML in first complete remission was investigated by flow cytometry. Patients who had HG levels more than 0.01% had a significantly better median LFS (29.2 vs 11.7 months; P = .001) and overall survival (not reached vs 23.5 months; P = .011). According to Cox analysis, an HG level more than 0.01% was an independent predictor of LFS (hazard ratio = 0.5; 95% confidence interval, 0.28-0.90, P < .03) when age, leukocytosis, the number of chemotherapy cycles, and the standardized cytogenetic and molecular risk subgroups were controlled for. These results indicate that HG analysis may help to define the risk of relapse in AML patients.
Identification of an Ire1alpha endonuclease specific inhibitor with cytotoxic activity against human multiple myeloma
Authors: Papandreou Ioanna , Denko Nicholas C. , Olson Michael , Van Melckebeke Heleen , Lust Sofie , Tam Arvin , Solow-Cordero David E. , Bouley Donna M. , Offner Fritz , Niwa Maho , Koong Albert C.
Published: November 2010

Activation of the adaptive Ire1-XBP1 pathway has been identified in many solid tumors and hematologic malignancies, including multiple myeloma (MM). Here, we report the identification of STF-083010, a novel small-molecule inhibitor of Ire1. STF-083010 inhibited Ire1 endonuclease activity, without affecting its kinase activity, after endoplasmic reticulum stress both in vitro and in vivo. Treatment with STF-083010 showed significant antimyeloma activity in model human MM xenografts. Similarly, STF-083010 was preferentially toxic to freshly isolated human CD138+ MM cells compared with other similarly isolated cell populations. The identification of this novel Ire1 inhibitor supports the hypothesis that the Ire1-XBP1 axis is a promising target for anticancer therapy, especially in the context of MM.
Comparative physical maps derived from BAC end sequences of tilapia (Oreochromis niloticus)
Authors: Soler Lucile , Conte Matthew A , Katagiri Takayuki , Howe Aimee E , Lee Bo-Young , Amemiya Chris , Stuart Andrew , Dossat Carole , Poulain Julie , Johnson Jeremy , Di Palma Federica , Lindblad-Toh Kerstin , Baroiller Jean-Francois , D'Cotta Helena , Ozouf-Costaz Catherine , Kocher Thomas D
Published: November 2010

Background

The Nile tilapia is the second most important fish in aquaculture. It is an excellent laboratory model, and is closely related to the African lake cichlids famous for their rapid rates of speciation. A suite of genomic resources has been developed for this species, including genetic maps and ESTs. Here we analyze BAC end-sequences to develop comparative physical maps, and estimate the number of genome rearrangements, between tilapia and other model fish species.

Results

We obtained sequence from one or both ends of 106,259 tilapia BACs. BLAST analysis against the genome assemblies of stickleback, medaka and pufferfish allowed identification of homologies for approximately 25,000 BACs for each species. We calculate that rearrangement breakpoints between tilapia and these species occur about every 3 Mb across the genome. Analysis of 35,000 clones previously assembled into contigs by restriction fingerprints allowed identification of longer-range syntenies.

Conclusions

Our data suggest that chromosomal evolution in recent teleosts is dominated by alternate loss of gene duplicates, and by intra-chromosomal rearrangements (~one per million years). These physical maps are a useful resource for comparative positional cloning of traits in cichlid fishes. The paired BAC end sequences from these clones will be an important resource for scaffolding forthcoming shotgun sequence assemblies of the tilapia genome.
Rapid induction of IgE responses to a worm cysteine protease during murine pre-patent schistosome infection
Authors: de Oliveira Fraga Lucia A , Lamb Erika W , Moreno Elizabeth C , Chatterjee Mitali , Dvořák Jan , Delcroix Melaine , Sajid Mohammed , Caffrey Conor R , Davies Stephen J
Published: November 2010

Background

During the pre-patent stage of infection, juvenileSchistosomablood flukes co-opt signals from the adaptive immune system to facilitate parasite development, but the types of responses that are induced at this early stage of infection, and the parasite antigens they target, have not been characterized.

Results

Through analysis of experimental pre-patent infections, we show that theS. mansonicysteine protease SmCB1 is rapidly targeted by an antigen-specific IgE response. The induction of this response is independent of schistosome eggs as infection with male or female worms alone also induced SmCB1-specific IgE. We also show that the SmCB1-specific IgE response is dependent on cognate CD4+T cell help and IL-4, suggesting that pre-patent Th2 responses provide T cell help for the SmCB1-specific IgE response. Finally, exposed human subjects also produced IgE against SmCB1.

Conclusions

Our data demonstrate that, like eggs, schistosome worms also induce functional type 2 responses and that a parasite cysteine protease is an inducer of type 2 responses during the early stages of schistosome infection.
A secreted serine protease of Paracoccidioides brasiliensis and its interactions with fungal proteins
Authors: Parente Juliana A , Salem-Izacc Sílvia M , Santana Jaime M , Pereira Maristela , Borges Clayton L , Bailão Alexandre M , Soares Célia MA
Published: November 2010

Background

Paracoccidioides brasiliensis is a thermodimorphic fungus, the causative agent of paracoccidioidomycosis (PCM). Serine proteases are widely distributed and this class of peptidase has been related to pathogenesis and nitrogen starvation in pathogenic fungi.

Results

A cDNA (Pb sp) encoding a secreted serine protease (Pb SP), was isolated from a cDNA library constructed with RNAs of fungal yeast cells recovered from liver of infected mice. Recombinant Pb SP was produced in Escherichia coli, and used to develop polyclonal antibodies that were able to detect a 66 kDa protein in the P. brasiliensis proteome. In vitro deglycosylation assays with endoglycosidase H demonstrated that Pb SP is a N-glycosylated molecule. The Pb sp transcript and the protein were induced during nitrogen starvation. The Pb sp transcript was also induced in yeast cells infecting murine macrophages. Interactions of Pb SP with P. brasiliensis proteins were evaluated by two-hybrid assay in the yeast Saccharomyces cerevisiae. Pb SP interacts with a peptidyl prolyl cis-trans isomerase, calnexin, HSP70 and a cell wall protein PWP2.

Conclusions

A secreted subtilisin induced during nitrogen starvation was characterized indicating the possible role of this protein in the nitrogen acquisition. Pb SP interactions with other P. brasiliensis proteins were reported. Proteins interacting with Pb SP are related to folding process, protein trafficking and cytoskeleton reorganization.
Functional analysis of the Theobroma cacao NPR1 gene in arabidopsis
Authors: Shi Zi , Maximova Siela N , Liu Yi , Verica Joseph , Guiltinan Mark J
Published: November 2010

Background

The Arabidopsis thaliana NPR1 gene encodes a transcription coactivator (NPR1) that plays a major role in the mechanisms regulating plant defense response. After pathogen infection and in response to salicylic acid (SA) accumulation, NPR1 translocates from the cytoplasm into the nucleus where it interacts with other transcription factors resulting in increased expression of over 2000 plant defense genes contributing to a pathogen resistance response.

Results

A putative Theobroma cacao NPR1 cDNA was isolated by RT-PCR using degenerate primers based on homologous sequences from Brassica, Arabidopsis and Carica papaya. The cDNA was used to isolate a genomic clone from Theobroma cacao containing a putative TcNPR1 gene. DNA sequencing revealed the presence of a 4.5 kb coding region containing three introns and encoding a polypeptide of 591 amino acids. The predicted TcNPR1 protein shares 55% identity and 78% similarity to Arabidopsis NPR1, and contains each of the highly conserved functional domains indicative of this class of transcription factors (BTB/POZ and ankyrin repeat protein-protein interaction domains and a nuclear localization sequence (NLS)). To functionally define the TcNPR1 gene, we transferred TcNPR1 into an Arabidopsis npr1 mutant that is highly susceptible to infection by the plant pathogen Pseudomonas syringae pv. tomato DC3000. Driven by the constitutive CaMV35S promoter, the cacao TcNPR1 gene partially complemented the npr1 mutation in transgenic Arabidopsis plants, resulting in 100 fold less bacterial growth in a leaf infection assay. Upon induction with SA, TcNPR1 was shown to translocate into the nucleus of leaf and root cells in a manner identical to Arabidopsis NPR1. Cacao NPR1 was also capable of participating in SA-JA signaling crosstalk, as evidenced by the suppression of JA responsive gene expression in TcNPR1 overexpressing transgenic plants.

Conclusion

Our data indicate that the TcNPR1 is a functional ortholog of Arabidopsis NPR1, and is likely to play a major role in defense response in cacao. This fundamental knowledge can contribute to breeding of disease resistant cacao varieties through the application of molecular markers or the use of transgenic strategies.
An in silico analysis of the mitochondrial protein import apparatus of plants
Authors: Carrie Chris , Murcha Monika W , Whelan James
Published: November 2010

Background

An in silico analysis of the mitochondrial protein import apparatus from a variety of species; including Chlamydomonas reinhardtii, Chlorella variabilis, Ectocarpus siliculosus, Cyanidioschyzon merolae, Physcomitrella patens, Selaginella moellendorffii, Picea glauca, Oryza sativa and Arabidopsis thaliana was undertaken to determine if components differed within and between plant and non-plant species.

Results

The channel forming subunits of the outer membrane components Tom40 and Sam50 are conserved between plant groups and other eukaryotes. In contrast, the receptor component(s) in green plants, particularly Tom20, (C. reinhardtii, C. variabilis, P. patens, S. moellendorffii, P. glauca, O. sativa and A. thaliana) are specific to this lineage. Red algae contain a Tom22 receptor that is orthologous to yeast Tom22. Furthermore, plant mitochondrial receptors display differences between various plant lineages. These are evidenced by distinctive motifs in all plant Metaxins, which are absent in red algae, and the presence of the outer membrane receptor OM64 in Angiosperms (rice and Arabidopsis), but not in lycophytes (S. moellendorffii) and gymnosperms (P. glauca). Furthermore, although the intermembrane space receptor Mia40 is conserved across a wide phylogenetic range, its function differs between lineages. In all plant lineages, Tim17 contains a C-terminal extension, which may act as a receptor component for the import of nucleic acids into plant mitochondria.

Conclusions

It is proposed that the observed functional divergences are due to the selective pressure to sort proteins between mitochondria and chloroplasts, resulting in differences in protein receptor components between plant groups and other organisms. Additionally, diversity of receptor components is observed within the plant kingdom. Even when receptor components are orthologous across plant and non-plant species, it appears that the functions of these have expanded or diverged in a lineage specific manner.
Mining the bitter melon (momordica charantial.) seed transcriptome by 454 analysis of non-normalized and normalized cDNA populations for conjugated fatty acid metabolism-related genes
Authors: Yang Peizhen , Li Xiangjun , Shipp Matthew J , Shockey Jay M , Cahoon Edgar B
Published: November 2010

Background

Seeds of Momordica charantia (bitter melon) produce high levels of eleostearic acid, an unusual conjugated fatty acid with industrial value. Deep sequencing of non-normalized and normalized cDNAs from developing bitter melon seeds was conducted to uncover key genes required for biotechnological transfer of conjugated fatty acid production to existing oilseed crops. It is expected that these studies will also provide basic information regarding the metabolism of other high-value novel fatty acids.

Results

Deep sequencing using 454 technology with non-normalized and normalized cDNA libraries prepared from bitter melon seeds at 18 DAP resulted in the identification of transcripts for the vast majority of known genes involved in fatty acid and triacylglycerol biosynthesis. The non-normalized library provided a transcriptome profile of the early stage in seed development that highlighted the abundance of transcripts for genes encoding seed storage proteins as well as for a number of genes for lipid metabolism-associated polypeptides, including Δ12 oleic acid desaturases and fatty acid conjugases, class 3 lipases, acyl-carrier protein, and acyl-CoA binding protein. Normalization of cDNA by use of a duplex-specific nuclease method not only increased the overall discovery of genes from developing bitter melon seeds, but also resulted in the identification of 345 contigs with homology to 189 known lipid genes in Arabidopsis. These included candidate genes for eleostearic acid metabolism such as diacylglycerol acyltransferase 1 and 2, and a phospholipid:diacylglycerol acyltransferase 1-related enzyme. Transcripts were also identified for a novel FAD2 gene encoding a functional Δ12 oleic acid desaturase with potential implications for eleostearic acid biosynthesis.

Conclusions

454 deep sequencing, particularly with normalized cDNA populations, was an effective method for mining of genes associated with eleostearic acid metabolism in developing bitter melon seeds. The transcriptomic data presented provide a resource for the study of novel fatty acid metabolism and for the biotechnological production of conjugated fatty acids and possibly other novel fatty acids in established oilseed crops.
Significant association between polymorphism of the erythropoietin gene promoter and myelodysplastic syndrome
Authors: Ma Wanlong , Kantarjian Hagop , Zhang Ke , Zhang Xi , Wang Xiuqiang , Chen Clifford , Donahue Amber C , Zhang Zhong , Yeh Chen-Hsiung , O'Brien Susan , Garcia-Manero Guillermo , Caporaso Neil , Landgren Ola , Albitar Maher
Published: November 2010

Background

Myelodysplastic syndrome (MDS) may be induced by certain mutagenic environmental or chemotherapeutic toxins; however, the role of susceptibility genes remains unclear. The G/G genotype of the single-nucleotide polymorphism (SNP) rs1617640 in the erythropoietin (EPO) promoter has been shown to be associated with decreased EPO expression. We examined the association of rs1617640 genotype with MDS.

Methods

We genotyped the EPO rS1617640 SNP in 189 patients with MDS, 257 with acute myeloid leukemia (AML), 106 with acute lymphoblastic leukemia, 97 with chronic lymphocytic leukemia, 353 with chronic myeloid leukemia, and 95 healthy controls.

Results

The G/G genotype was significantly more common in MDS patients (47/187; 25.1%) than in controls (6/95; 6.3%) or in patients with other leukemias (101/813; 12.4%) (all P < 0.001). Individuals with the G/G genotype were more likely than those with other genotypes to have MDS (odd ratio = 4.98; 95% CI = 2.04-12.13). Clinical and follow up data were available for 112 MDS patients and 186 AML patients. There was no correlation between EPO promoter genotype and response to therapy or overall survival in MDS or AML. In the MDS group, the GG genotype was significantly associated with shorter complete remission duration, as compared with the TT genotype (P = 0.03). Time to neutrophils recovery after therapy was significantly longer in MDS patients with the G/G genotype (P = 0.02).

Conclusions

These findings suggest a strong association between the rs1617640 G/G genotype and MDS. Further studies are warranted to investigate the utility of screening for this marker in individuals exposed to environmental toxins or chemotherapy.
Tumour auto-antibody screening: performance of protein microarrays using SEREX derived antigens
Authors: Stempfer René , Syed Parvez , Vierlinger Klemens , Pichler Rudolf , Meese Eckart , Leidinger Petra , Ludwig Nicole , Kriegner Albert , Nöhammer Christa , Weinhäusel Andreas
Published: November 2010

Background

The simplicity and potential of minimal invasive testing using serum from patients make auto-antibody based biomarkers a very promising tool for use in diagnostics of cancer and auto-immune disease. Although several methods exist for elucidating candidate-protein markers, immobilizing these onto membranes and generating so called macroarrays is of limited use for marker validation. Especially when several hundred samples have to be analysed, microarrays could serve as a good alternative since processing macro membranes is cumbersome and reproducibility of results is moderate.

Methods

Candidate markers identified by SEREX (serological identification of antigens by recombinant expression cloning) screenings of brain and lung tumour were used for macroarray and microarray production. For microarray production recombinant proteins were expressed in E. coli by autoinduction and purified His-tag (histidine-tagged) proteins were then used for the production of protein microarrays. Protein arrays were hybridized with the serum samples from brain and lung tumour patients.

Result

Methods for the generation of microarrays were successfully established when using antigens derived from membrane-based selection. Signal patterns obtained by microarrays analysis of brain and lung tumour patients' sera were highly reproducible (R = 0.92-0.96). This provides the technical foundation for diagnostic applications on the basis of auto-antibody patterns. In this limited test set, the assay provided high reproducibility and a broad dynamic range to classify all brain and lung samples correctly.

Conclusion

Protein microarray is an efficient means for auto-antibody-based detection when using SEREX-derived clones expressing antigenic proteins. Protein microarrays are preferred to macroarrays due to the easier handling and the high reproducibility of auto-antibody testing. Especially when using only a few microliters of patient samples protein microarrays are ideally suited for validation of auto-antibody signatures for diagnostic purposes.
Utilization of delivery care among rural women in china: does the health insurance make a difference? a cross-sectional study
Authors: Xiao Shengbin , Yan Hong , Shen Yuan , Dang Shaonong , Hemminki Elina , Wang Duolao , Long Qian , Gao Jianmin
Published: November 2010

Background

Since 2003, the New Cooperative Medical Scheme (NCMS) has been implemented throughout rural China, usually covering delivery services in its benefit package. The objective of this study was to compare the difference of utilization of delivery services, expenditures, and local women's perceived affordability between women with and without reimbursement from NCMS.

Methods

A cross-sectional survey was carried out in two rural counties in Shaanxi province, China, during December 2008-March 2009. Women giving birth from April 2008 to March 2009 were interviewed by a structured questionnaire to collect information on utilization of delivery services. Multivariable analyses were used to compare the differences in outcomes between women with and without reimbursement from NCMS.

Results

Of the total 1613 women interviewed, 747(46.3%) got reimbursement to cover their expenditure on delivery care (NCMS group) and 866(53.7%) paid delivery services entirely out of their own pocket (Non-NCMS group). Compared with the Non-NCMS group, the NCMS group had significantly more women who delivered at hospital. The rate of Caesarean section (CS), proportion of women seeking higher level services, and length of hospitalization were similar between the two groups. The total hospital costs for delivery services in the NCMS group was significantly smaller and after being reimbursed, the out-of-pocket payment in the NCMS group was less than a half of that in the Non-NCMS group. Fewer women in the NCMS group than in the Non-NCMS group considered their payment for delivery services expensive.

Conclusions

There was no evidence of overuse delivery services among the women reimbursed by NCMS. Total hospital costs and women's costs for delivery services were found lower in the NCMS group, subsequently alleviation on women's perceived financial affordability.
DockAnalyse: an application for the analysis of protein-protein interactions
Authors: Amela Isaac , Delicado Pedro , Gómez Antonio , Bonàs Sílvia , Querol Enrique , Cedano Juan
Published: November 2010

Background

Is it possible to identify what the best solution of a docking program is? The usual answer to this question is the highest score solution, but interactions between proteins are dynamic processes, and many times the interaction regions are wide enough to permit protein-protein interactions with different orientations and/or interaction energies. In some cases, as in a multimeric protein complex, several interaction regions are possible among the monomers. These dynamic processes involve interactions with surface displacements between the proteins to finally achieve the functional configuration of the protein complex. Consequently, there is not a static and single solution for the interaction between proteins, but there are several important configurations that also have to be analyzed.

Results

To extract those representative solutions from the docking output datafile, we have developed an unsupervised and automatic clustering application, named DockAnalyse. This application is based on the already existing DBscan clustering method, which searches for continuities among the clusters generated by the docking output data representation. The DBscan clustering method is very robust and, moreover, solves some of the inconsistency problems of the classical clustering methods like, for example, the treatment of outliers and the dependence of the previously defined number of clusters.

Conclusions

DockAnalyse makes the interpretation of the docking solutions through graphical and visual representations easier by guiding the user to find the representative solutions. We have applied our new approach to analyze several protein interactions and model the dynamic protein interaction behavior of a protein complex. DockAnalyse might also be used to describe interaction regions between proteins and, therefore, guide future flexible dockings. The application (implemented in the R package) is accessible.
Laxative effects of agarwood on low-fiber diet-induced constipation in rats
Authors: Kakino Mamoru , Tazawa Shigemi , Maruyama Hiroe , Tsuruma Kazuhiro , Araki Yoko , Shimazawa Masamitsu , Hara Hideaki
Published: November 2010

Background

Agarwood (Aquilaria sinensis), well known as incense in Southeast Asia, has been used as a digestive in traditional medicine. We investigated the laxative effects of an ethanol extract of agarwood leaves (EEA) in a rat model of low-fiber diet-induced constipation.

Methods

A set of rats was bred on a normal diet while another set was placed on a low-fiber diet to induce constipation. The laxative effect of agarwood was then investigated on both sets of rats.

Results

Pretreatment of normal rats with single dose of EEA (600 mg/kg, p.o.) significantly increased frequency and weight of stools. Also, treatments with EEA (300 and 600 mg/kg, p.o.) for 14 days caused a significant increase in stool frequency and weight. Feeding of the animals with a low-fiber diet resulted in a decrease in stool weight, frequency, and water content and also delayed carmine egestion. A single treatment with EEA (600 mg/kg) or senna (150 and 300 mg/kg) significantly increased stool frequency, weight, and water content and also accelerated carmine egestion in the model rats. Once daily administrations of EEA (150 mg/kg), for 14 days, caused a significant increase in water content of stools. The higher doses of EEA (300 and 600 mg/kg) significantly increased frequency, weight, and water content of the stools while accelerating carmine egestion in the constipated rats. Senna (150 and 300 mg/kg) produced similar effect as the higher doses of EEA but, in addition, induced severe diarrhea.

Conclusion

These findings indicate that EEA has a laxative effect, without causing diarrhea, in a rat model of low-fiber diet-induced constipation. These findings suggest that EEA may be highly effective on constipation as a complementary medicine in humans suffering from life style-induced constipation.
Evaluation of computerized health management information system for primary health care in rural India
Authors: Krishnan Anand , Nongkynrih Baridalyne , Yadav Kapil , Singh Satyavir , Gupta Vivek
Published: November 2010

Background

The Comprehensive Rural Health Services Project Ballabgarh, run by All India Institute of Medical Sciences (AIIMS), New Delhi has a computerized Health Management Information System (HMIS) since 1988. The HMIS at Ballabgarh has undergone evolution and is currently in its third version which uses generic and open source software. This study was conducted to evaluate the effectiveness of a computerized Health Management Information System in rural health system in India.

Methods

The data for evaluation were collected by in-depth interviews of the stakeholders i.e. program managers (authors) and health workers. Health Workers from AIIMS and Non-AIIMS Primary Health Centers were interviewed to compare the manual with computerized HMIS. A cost comparison between the two methods was carried out based on market costs. The resource utilization for both manual and computerized HMIS was identified based on workers' interviews.

Results

There have been no major hardware problems in use of computerized HMIS. More than 95% of data was found to be accurate. Health workers acknowledge the usefulness of HMIS in service delivery, data storage, generation of workplans and reports. For program managers, it provides a better tool for monitoring and supervision and data management. The initial cost incurred in computerization of two Primary Health Centers was estimated to be Indian National Rupee (INR) 1674,217 (USD 35,622). Equivalent annual incremental cost of capital items was estimated as INR 198,017 (USD 4213). The annual savings is around INR 894,283 (USD 11,924).

Conclusion

The major advantage of computerization has been in saving of time of health workers in record keeping and report generation. The initial capital costs of computerization can be recovered within two years of implementation if the system is fully operational. Computerization has enabled implementation of a good system for service delivery, monitoring and supervision.
Insect cells are superior to Escherichia coli in producing malaria proteins inducing IgG targeting PfEMP1 on infected erythrocytes
Authors: Victor Michala E , Bengtsson Anja , Andersen Gorm , Bengtsson Dominique , Lusingu John P , Vestergaard Lasse S , Arnot David E , Theander Thor G , Joergensen Louise , Jensen Anja TR
Published: November 2010

Background

The PFD1235wPlasmodium falciparumerythrocyte membrane protein 1 (PfEMP1) antigen is associated with severe malaria in children and can be expressed on the surface of infected erythrocytes (IE) adhering to ICAM1. However, the exact three-dimensional structure of this PfEMP1 and its surface-exposed epitopes are unknown. An insect cell andEscherichia colibased system was used to express single and double domains encoded by thepfd1235w vargene. The resulting recombinant proteins have been evaluated for yield and purity and their ability to induce rat antibodies, which react with the native PFD1235w PfEMP1 antigen expressed on 3D7PFD1235w-IE. Their recognition by human anti-malaria antibodies from previously infected Tanzanian donors was also analysed.

Methods

The recombinant proteins were run on SDS-PAGE and Western blots for quantification and size estimation. Insect cell andE. coli-produced recombinant proteins were coupled to a bead-based Luminex assay to measure the plasma antibody reactivity of 180 samples collected from Tanzanian individuals. The recombinant proteins used for immunization of rats and antisera were also tested by flow cytometry for their ability to surface label 3D7PFD1235w-IE.

Results

All seven pAcGP67A constructs were successfully expressed as recombinant protein in baculovirus-infected insect cells and subsequently produced to a purity of 60-97% and a yield of 2-15 mg/L. By comparison, only three of seven pET101/D-TOPO constructs expressed in theE. colisystem could be produced at all with purity and yield ranging from 3-95% and 6-11 mg/L. All seven insect cell, but only two of theE. coliproduced proteins induced antibodies reactive with native PFD1235w expressed on 3D7PFD1235w-IE. The recombinant proteins were recognized in an age- and transmission intensity-dependent manner by antibodies from 180 Tanzanian individuals in a bead-based Luminex assay.

Conclusions

The baculovirus based insect cell system was distinctly superior to theE. coliexpression system in producing a larger number of different recombinant PFD1235w protein domains and these were significantly easier to purify at a useful yield. However, proteins produced in both systems were able to induce antibodies in rats, which can recognize the native PFD1235w on the surface of IE.
Augmented plasma microparticles during acute Plasmodium vivax infection
Authors: Campos Fernanda MF , Franklin Bernardo S , Teixeira-Carvalho Andréa , Filho Agnaldo LS , de Paula Sálua CO , Fontes Cor J , Brito Cristiana F , Carvalho Luzia H
Published: November 2010

Background

In the last few years, the study of microparticles (MPs) - submicron vesicles released from cells upon activation or apoptosis - has gained growing interest in the field of inflammation and in infectious diseases. Their role in the human malaria parasite Plasmodium vivax remains unexplored. Because acute vivax malaria has been related to pro-inflammatory responses, the main hypothesis investigated in this study was that Plasmodium vivax infection is associated with elevated levels of circulating MPs, which may play a role during acute disease in non-immune patients.

Methods

Plasma MPs were analysed among thirty-seven uncomplicated P. vivax infections from an area of unstable malaria transmission in the Brazilian Amazon. The MP phenotype was analysed by flow cytometry using the classical MP marker, annexin, and fluorochrome-labeled monoclonal antibodies against specific cell surface markers. The frequencies of plasma MPs in P. vivax patients (n = 37) were further compared to malaria-unexposed controls (n = 15) and ovarian carcinoma patients (n = 12), a known MPs-inducing disease non-related to malaria.

Results

The frequencies of plasma circulating MPs were markedly increased in P. vivax patients, as compared to healthy age-matched malaria-unexposed controls. Although platelets, erythrocytes and leukocytes were the main cellular sources of MPs during vivax malaria, platelet derived-MPs (PMPs) increased in a linear fashion with the presence of fever at the time of blood collection (β = 0.06, p < 0.0001) and length of acute symptoms (β = 0.36, p < 0.0001). Finally, the results suggest that plasma levels of PMPs diminish as patient experience more episodes of clinical malaria (β = 0.07, p < 0.003).

Conclusions

Abundant circulating MPs are present during acute P. vivax infection, and platelet derived-MPs may play a role on the acute inflammatory symptoms of malaria vivax.