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3.
J Surg Res ; 163(1): 12-7, 2010 Sep.
Article in English | MEDLINE | ID: mdl-20599226

ABSTRACT

BACKGROUND: Calcific aortic stenosis (CAS) is the most common acquired valvular disorder in industrialized countries. This study investigates the correlation of different known biomarkers for CAS as a first step towards the development of a panel of biomarkers that can be used in prognostic staging. METHODS: Venous blood samples were obtained from both patients with CAS scheduled for surgery and healthy individuals. Plasma levels of fetuin-A, NT-proBNP, BNP, homocysteine and osteopontin were measured by enzyme-linked immunosorbent assay (ELISA). CAS was measured by echocardiography and was defined as an aortic valve area of less than 2.0 cm(2). Non-paired t-tests were used for comparison. RESULTS: CAS was present in 33 subjects (mean age 75.9 y) and absent in 11 subjects (mean age 55.36 y). Individuals with CAS exhibited higher plasma levels of NT-proBNP (1.33 versus 0.73 pmol/mL, P < 0.05), BNP fragment (1.47 versus 0.34 ng/mL P < 0.05), and osteopontin (60.79 versus 25.42 ng/mL P < 0.05) compared with controls. Fetuin-A levels were lower in individuals with CAS than in healthy controls (0.25 versus 0.34g/L, P < 0.05). Asymmetric dimethylarginine (ADMA) were lower (1.08 versus 1.1 micromol/L, P > 0.05) while homocysteine levels (20.34 +/- 2.14 versus 19.23 +/- 4.19 P > 0.05) were higher in the CAS patients. CONCLUSION: This study demonstrates a direct correlation of NT-pro-BNP, BNP, and osteopontin and the presence of CAS, while fetuin A showed an inverse correlation. Plasma ADMA and homocysteine levels were comparable in the CAS patients and healthy individuals. This is the first study in which several biomarkers previously studied independently in patients with CAS have been investigated simultaneously in the same study population.


Subject(s)
Aortic Valve Stenosis/blood , Biomarkers/blood , Calcinosis/blood , Adult , Aged , Aged, 80 and over , Aortic Valve Stenosis/diagnosis , Calcinosis/diagnosis , Female , Humans , Male , Middle Aged
4.
Adv Genet ; 57: 49-96, 2007.
Article in English | MEDLINE | ID: mdl-17352902

ABSTRACT

A consortium of investigators is engaged in a functional genomics project centered on the filamentous fungus Neurospora, with an eye to opening up the functional genomic analysis of all the filamentous fungi. The overall goal of the four interdependent projects in this effort is to accomplish functional genomics, annotation, and expression analyses of Neurospora crassa, a filamentous fungus that is an established model for the assemblage of over 250,000 species of non yeast fungi. Building from the completely sequenced 43-Mb Neurospora genome, Project 1 is pursuing the systematic disruption of genes through targeted gene replacements, phenotypic analysis of mutant strains, and their distribution to the scientific community at large. Project 2, through a primary focus in Annotation and Bioinformatics, has developed a platform for electronically capturing community feedback and data about the existing annotation, while building and maintaining a database to capture and display information about phenotypes. Oligonucleotide-based microarrays created in Project 3 are being used to collect baseline expression data for the nearly 11,000 distinguishable transcripts in Neurospora under various conditions of growth and development, and eventually to begin to analyze the global effects of loss of novel genes in strains created by Project 1. cDNA libraries generated in Project 4 document the overall complexity of expressed sequences in Neurospora, including alternative splicing alternative promoters and antisense transcripts. In addition, these studies have driven the assembly of an SNP map presently populated by nearly 300 markers that will greatly accelerate the positional cloning of genes.


Subject(s)
Neurospora/genetics , Base Sequence , Chromosome Mapping , DNA, Fungal/genetics , Gene Deletion , Gene Expression Profiling , Gene Library , Genetic Techniques , Genome, Fungal , Genomics , Mutation , Oligonucleotide Array Sequence Analysis , Phenotype , Polymorphism, Single Nucleotide
5.
Proc Natl Acad Sci U S A ; 103(27): 10352-10357, 2006 Jul 05.
Article in English | MEDLINE | ID: mdl-16801547

ABSTRACT

The low rate of homologous recombination exhibited by wild-type strains of filamentous fungi has hindered development of high-throughput gene knockout procedures for this group of organisms. In this study, we describe a method for rapidly creating knockout mutants in which we make use of yeast recombinational cloning, Neurospora mutant strains deficient in nonhomologous end-joining DNA repair, custom-written software tools, and robotics. To illustrate our approach, we have created strains bearing deletions of 103 Neurospora genes encoding transcription factors. Characterization of strains during growth and both asexual and sexual development revealed phenotypes for 43% of the deletion mutants, with more than half of these strains possessing multiple defects. Overall, the methodology, which achieves high-throughput gene disruption at an efficiency >90% in this filamentous fungus, promises to be applicable to other eukaryotic organisms that have a low frequency of homologous recombination.


Subject(s)
Fungal Proteins/metabolism , Gene Deletion , Mutagenesis, Insertional/methods , Neurospora/genetics , Neurospora/metabolism , Transcription Factors/deficiency , Transcription Factors/metabolism , DNA Primers/genetics , Fungal Proteins/genetics , Gene Expression Regulation, Fungal , Hyphae , Mutagenesis, Insertional/genetics , Mutation/genetics , Neurospora/growth & development , Phenotype , Transcription Factors/genetics
6.
Biochim Biophys Acta ; 1760(6): 848-57, 2006 Jun.
Article in English | MEDLINE | ID: mdl-16574329

ABSTRACT

N. crassa has two forms of arginase. The physiological role of multiple arginases is not understood. The two forms were shown to be differentially expressed from a single locus (aga) and both proteins are localized to the cytoplasm. The 36-kDa protein was expressed in minimal and arginine supplemented medium, whereas the 41-kDa form was detected only in the presence of arginine. In this study we examined developmental expression of the two arginase transcripts and proteins in conidia and during conidial germination. Two novel observations are revealed, storage of both arginase proteins in conidia and temporal expression of aga transcripts during early germination. To better understand the role of arginase in conidia and the nature of the temporal expression, we examined the effects of related metabolites, arginine, ornithine, proline, glutamate and glutamine on protein storage and temporal expression. These metabolites were used as supplements or sole nitrogen sources. Storage of arginase protein was detected in all conidial samples examined except when glutamate was used as the nitrogen source. The aga temporal RNA expression early in germination was abolished when arginine related metabolites were used as nitrogen sources. The exception to this result is observed with glutamate where temporal expression was seen when glutamate was the sole nitrogen source and abolished with glutamate supplementation. The temporal expression result supports a unique role for arginase in glutamate accumulation early in germination whereas the protein storage result supports the existence of a novel pathway utilizing arginase for glutamate synthesis in asexual spore development.


Subject(s)
Arginase/genetics , Arginase/metabolism , Gene Expression Regulation, Fungal , Neurospora crassa/enzymology , Neurospora crassa/growth & development , Arginine/chemistry , Arginine/pharmacology , Culture Media , Glutamic Acid/pharmacology , Isoenzymes/genetics , Isoenzymes/metabolism , Neurospora crassa/genetics , Ornithine/pharmacology , Proline/pharmacology , RNA, Messenger/genetics , RNA, Messenger/metabolism , Time Factors
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