Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts:   Target Concepts:
Query: UNIPROT:P06889 (Mol)
 630,302 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

A seminested polymerase chain reaction (PCR)-based diagnostic assay was evaluated for detection and verification of Neisseria meningitidis, Haemophilus influenzae, Streptococcus pneumoniae, Steptococcus agalactiae and Listeria monocytogenes in cerebrospinal fluid (CSF) and other biological samples. A general bacterial amplicon from the 16S rRNA gene was amplified in a first step, and species-specific regions in a second. The detection level was 4 fg DNA/reaction, corresponding to about one bacterial genome per reaction tube. Sample preparations (Dynabeads DNA DIRECT kit) were assayed from 140 bacterial strains suspended in saline. In CSF the detection level for bacteria was 10(3)CFU ml-1for N. meningitidis, H. influenzae and S. pneumoniae, 10(4)CFU ml-1for Escherichia coli and 10(5)CFU ml-1for S. agalactiae and L. monocytogenes. The detection levels for these bacteria were the same in the other tested biological samples, like blood with or without culture media. Clinical CSF samples were evaluated from 71 patients with proven bacterial meningitis, as were 61 CSF samples from individuals without bacterial meningitis. The diagnostic sensitivity of the assay in detecting bacteria in general was 0.97, and for the specific species in the clinical CSF samples 0.87-0.94. The specificity was 1.0 for detecting bacteria in general. Some cross-reactions were noted within the streptococcus group. The PCR results were verified by banding patterns of Hae III digested PCR products.
Mol Cell Probes 1999 Feb
PMID:Evaluation of an extended diagnostic PCR assay for detection and verification of the common causes of bacterial meningitis in CSF and other biological samples. 1002 33

Using ouabain sensitive 86Rb uptake by the vessel wall, we previously showed that sodium-potassium pump activity is decreased in the arteries and veins, and that the sodium-potassium pump inhibitor (SPI) is increased in the plasma of dogs with one-kidney, one wrap (1-K, 1W) hypertension, a low renin model of hypertension. We also showed in rats with a similar type of hypertension that the membrane potential of vascular smooth muscle cells in arteries is decreased, and that this decrease can be reproduced in arterial cells in arteries from normal rats by applying plasma from the hypertensive animals. One endogenous SPI in human plasma has been reported to be ouabain or its isomer. In this study, we used a newly available Dupont ouabain enzyme immunoassay kit to examine plasma and kidneys for SPI in dogs with 1-K, 1W hypertension. We also examined 1) the inhibiting activity of plasma of Na+, K(+)-ATPase obtained from normal kidneys, and 2) the Na+, K(+)-ATPase activity of the kidneys from these hypertensive animals. 1-K, 1W hypertension was produced in dogs by wrapping the left kidney in a silk bag and removing the right kidney. The removed kidney was kept at -70 degrees C till assayed. After 4 weeks of hypertension, the remaining kidney was removed and stored at -70 degrees C till assayed. Blood samples were drawn before and at weeks 3 and 4 of hypertension. Plasma levels of "ouabain" and Na+, K(+)-ATPase inhibitory activity were increased at weeks 3 and 4 of hypertension, compared to pre-hypertension levels. Renal tissue "ouabain" levels were also increased at week 4 of hypertension. However, renal Na+, K(+)-ATPase activity was unchanged. These findings, using two different assays, confirm our 1980 conclusion that SPI is elevated in the plasma of dogs with 1-K, 1W hypertension. The absence of renal Na+, K(+)-ATPase inhibition, despite increased plasma and renal SPI in these animals, may have important implications for the development of this type of hypertension.
Cell Mol Biol (Noisy-le-grand) 1999 Feb
PMID:Sodium-potassium pump inhibitor in the mechanism of one-kidney, one wrap hypertension in dogs. 1009 45

The mother and second child from a family, already with one PI ZZ child, were typed PI MZ by isoelectric focusing and unexpectedly as PI ZZ using a commercial alpha-1-antitrypsin genotyping kit. Both methods typed the father and first child as PI MZ and PI ZZ, respectively. DNA sequence analysis identified a 26-base pair (bp) deletion and 2-bp insertion in intron IV of the normal PI*M allele from both the mother and second child. The majority of the binding site for an amplification primer of the genotyping kit was absent in the variant deletion-insertion allele. The apparent PI*Z/PI*Z genotype of the mother and second child therefore arose from amplification of the PI*Z allele alone. Two hundred random DNA samples were subsequently examined and 5 of these were found to be heterozygous for the same deletion-insertion allele. The authors have designated the previously undescribed PI*M allele that harbors this benign polymorphism PI*Mwhitstable. The genotyping kit has been redesigned and revalidated, and its performance is not affected by the presence of the PI*Mwhitstable allele. The Gen Bank accession number for the nucleotide sequence described is AF159454.
Diagn Mol Pathol 1999 Dec
PMID:Molecular characterization of a new alpha-1-antitrypsin M variant allele, Mwhitstable: implications for DNA-based diagnosis. 1061 77

Non-human primates (NHPs) are increasingly utilized as models to investigate different aspects of immune responses against self (autoimmunity) and foreign antigens. These animals provide valuable models for testing the efficacy of candidate vaccines against pathogens such as human immunodeficiency virus (HIV) and also fertility regulating agents (immunocontraceptives). In order to fully understand the effects of vaccination, it may be necessary to elucidate the immunogenetic background of these animals. The major histocompatibility complex (Mhc) molecules play an important role in the generation of effective immune responses. Serological techniques have been used in the identification of human leukocyte antigens (HLA) necessary for cross-matching organs and tissues for transplantation. However, the application of this technique for typing monkey Mhc alleles has been hampered by unavailability of well characterized immunological reagents. Polymerase chain reaction (PCR)-based techniques such as restriction fragment length polymorphism (RFLP) and sequence-specific oligonucleotide probe hybridization (SSOP) have been extensively used for typing HLA-DP, DQ and DR alleles. A commercially available Kit (AmpliTypeR) designed for amplification and typing of HLA DQalpha alleles is routinely used in typing DNA samples for forensic casework. In the present study, we have evaluated this kit for possible application in routine typing of primate DQA1 alleles. Genomic DNA from ten African primate species (23 individuals) was isolated from peripheral blood lymphocytes and polymorphic second exon of DQA1 locus amplified using GH26 and GH27 PCR primers. The PCR products were hybridized on a nylon membrane containing immobilized sequence-specific oligonucleotide probes. Our results show seven of the nine probes hybridizing with primate DQA1 alleles, indicating that typing of equivalent primate alleles can be accomplished at lower stringency conditions. However, it may be necessary to design additional oligonucleotides probes (based on available primate DQA1 sequences) to improve the discriminating power of this kit for use in routine typing of Old World monkey DQA1 alleles.
Cell Mol Biol (Noisy-le-grand) 1999 Dec
PMID:DNA typing of primate major histocompatibility complex (Mhc)-DQA1 locus by PCR and dot blot hybridization. 1064 74

We describe a simple and rapid method for cloning insect vitellogenin (Vg) cDNAs. The method relies on the facts that insect Vg amino acid sequences can be aligned confidently along their entire lengths and that a short, highly conserved GL/ICG motif and up to nine cysteine residues that follow at conserved locations are present near the C-termini. An adaptor-ligated double-strand cDNA library is constructed from poly(A)+ RNA prepared from vitellogenic female fat body tissues using a commercial kit, and subjected to PCR with each of the degenerate nucleotide sequences for the GL/ICG motif and the adaptor sequence as primers. The PCR products (0.7-0.9 kb, representing the 3' portion) are cloned, the nucleotide sequences are determined, and the deduced amino acid sequences are aligned with the known insect Vg sequences starting from the GL/ICG motif. Gene-specific primers corresponding to the sequences near the 5'-termini of the initial clones and the adaptor sequence are employed to obtain the remaining 5' portion of the Vg cDNAs. The method was successfully applied to the bean bug Plautia stali (Heteroptera), revealing three Vg genes.
Insect Biochem Mol Biol 2000 Mar
PMID:A simple and rapid method for cloning insect vitellogenin cDNAs. 1073 86

Since Flegg (H.M. Flegg, An investigation of the determination of serum cholesterol by an enzymatic method, Ann. Clin. Biochem. 10 (1973) 79-84) and Richmond (W. Richmond, The development of an enzymatic technique for the assay of cholesterol in biological fluids, Scand. J. clin. Lab. Invest. 29 (1972) 25; W. Richmond, Preparation and properties of a bacterial cholesterol oxidase from Nocardia sp. and its application to enzyme assay of total cholesterol in serum, Clinical Chemistry 19 (1973) 1350-1356) first illustrated the suitability of cholesterol oxidase (COD) for the analysis of serum cholesterol, COD has risen to become the most widely used enzyme in clinical laboratories with the exception of glucose oxidase (GOD). The use is widespread because assays incorporating the enzyme are extremely simple, specific, and highly sensitive and thus offer distinct advantages over the Liebermann-Burchard analytical methodologies which employ corrosive reagents and can be prone to unreliable results due to interfering substances such as bilirubin. Individuals can now readily determine their own serum cholesterol levels with a simple disposable test kit. This review discusses COD in some detail and includes the topics: (1) The variety of bacterial sources available; (2) The various extraction/purification protocols utilised in order to obtain protein of sufficient clarification (purity) for use in food/clinical analysis; (3) Significant differences in the properties of the individual enzymes; (4) Substrate specificities of the various enzymes; (5) Examples of biological assays which have employed cholesterol oxidase as an integral part of the analysis, and the various assay protocols; (6) New steroidal products of COD. This review is not a comprehensive description of published work, but is intended to provide an account of recent and current research, and should promote further interest in the application of enzymes to analytical selectivity.
J Steroid Biochem Mol Biol 2000 Apr
PMID:Cholesterol oxidase: sources, physical properties and analytical applications. 1082 8

Although serum prostate specific antigen (PSA), derived from cellular PSA through secretion, is widely used as a marker for prostate cancer (CAP), the exact regulatory mechanism of its secretion is not fully understood. To explore the regulation of serum PSA concentration, we examined whether the glycosylation state of cellular PSA might be associated with its secretion, thus determining its serum concentration. Blood and prostate tissue specimens were obtained from patients undergoing radical prostatectomy. Following preparation of cell extracts by tissue homogenization, the concentrations of serum and cellular PSA were determined using the Tandem-E PSA kit. The extent of cellular PSA glycosylation was then assessed by Western blot and affinoblott analyses. Neither serum nor cellular PSA concentrations correlated with the Gleason scores. Similarly, no direct relation between serum and cellular PSA levels was observed. However, the Western blots showed that the cellular PSA proteins were converted to the deglycosylated forms with glycosidase treatment, indicating differential glycosylation of cellular PSA. Affinoblotting further revealed that the various amounts of PSA glycosylation were associated wtih the serum PSA levels, with an inverse correlation between serum PSA and cellular PSA glycosylation: the greater the PSA glycosylation, the lower the serum PSA, and vice versa. The present study demonstrates that cellular PSAs in CAP specimens are differentially glycosylated and that such difference correlates well with the serum PSA concentration. Therefore, the concentrations of serum PSA appear to depend in part on a selective secretion of cellular PSA, which could be regulated primarily by its glycosylation state.
Mol Urol 1999
PMID:Differential Glycosylation of Cellular Prostate Specific Antigen and the Regulatory Mechanism of Its Secretion. 1085 17

It was reported that a hamster protein, called "oscillin," with a sequence related to that of an Escherichia coli GNPDA triggered Ca(2+) oscillations in mammalian oocytes when introduced into their cytoplasm upon fertilization. Recently, it was shown that GNPDA/oscillin is ubiquitously expressed in rat tissues and that a recombinant hamster GNPDA/oscillin protein does not exhibit oscillin activity when injected into oocytes. In the mouse, the nature and role of such a GNPDA/oscillin is not known, but another candidate protein, tr-kit, has been proposed as a sperm factor causing oocyte activation. In order to clarify this issue, we have characterized the mouse homolog of hamster and human GNPDA/oscillin, and examined its expression along with that of tr-kit, in parallel. We report here the molecular cloning and sequencing of mouse GNPDA/oscillin, which shows over 96% identity with the hamster and human homologs. Using specific primers, we performed an RT-PCR analysis to determine the tissue distribution of mouse GNPDA/oscillin mRNA. Unlike tr-kit mRNA which is expressed solely in mouse testis, GNPDA/oscillin mRNA is detected in unfertilized oocytes and in all tissues examined including testis, heart, thymus, liver, ovary, uterus, kidney, spleen, and lung. The protein itself is also detected in all tissues examined by Western blots. Indirect immunofluorescence studies, using an antibody raised against hamster GNPDA, demonstrate that GNPDA is lost with the acrosome reaction of mouse spermatozoa, is localized in the equatorial and neck regions of the human spermatozoa and the post-acrosomal region of the hamster spermatozoa. Our results thus indicate that mouse GNPDA/oscillin, the homolog of hamster oscillin, unlike tr-kit, does not exhibit some of the required characteristics expected from a putative sperm-derived oocyte-activating factor.
Mol Reprod Dev 2000 Jul
PMID:Cloning, sequencing, and expression analysis of mouse glucosamine-6-phosphate deaminase (GNPDA/oscillin). 1086 10

We have investigated the distribution of immunoreactive endothelins (irET) in fetal fluids and expression of ET precursor genes in villous tissue during the first trimester. Samples of maternal plasma (n = 6), coelomic fluid (n = 28), amniotic fluid (n = 23) and villous tissue (n = 3) were obtained from 30 pregnancies immediately before surgical termination at 7-12 weeks gestation. irET concentration was measured in plasma and fluids using two different radioimmunoassay kits, i.e. RPA 545 and RPA 555 and high performance liquid chromatography (HPLC). Total RNA was extracted and purified from villous tissue, reverse transcription and polymerase chain reaction (RT-PCR) were performed to evaluate the expression of ET-related genes. The irET concentration as evaluated by both kits was significantly higher (P<0.005) in maternal plasma than in coelomic or amniotic fluid and significantly higher (P<0.005) in coelomic fluid than in amniotic fluid using the RPA 555 kit. The profile of ET obtained by the HPLC- radioimmunoassay (RPA 555 kit) method confirmed significantly (P<0.005) higher ET concentration in coelomic than in amniotic fluid, although a similar distribution pattern for the three ET was observed in both embryonic fliud cavities. ET-3 was the predominant isoform in both fluids, reaching 19.4+/-2.0 pg/ml and 6.3+/-1.6 pg/ml in coelomic and amniotic fluid, respectively. Coelomic or amniotic fluid irET concentration did not change with gestational age irrespective of the kit used. RT-PCR demonstrated that first trimester placenta expresses the genes encoding for prepro-ET-1, -ET-2 and -ET-3. The similar ET distribution pattern in both fluid cavities could reflect their origin from the villous tissue and suggests that ET may play a role in the development of placenta and other fetal organs during organogenesis.
Mol Hum Reprod 2000 Aug
PMID:Placental endothelin gene expression and endothelin concentration in fetal fluids of the first trimester gestational sac. 1090 87

BSE, first identified in the UK in 1986 is thought to have arisen from feeding scrapie infected Meat and Bone Meal (MBM), produced under sub-optimal conditions, to cattle. For quality and safety reasons there is a requirement for a good analytical test for the surveillance of processed MBM. This study describes species-specific PCR assays for the identification of ovine, porcine and poultry species in MBM. A comparison between two distinct DNA extraction methods, i.e. the silicaguanidiumthiocyanate DNA isolation procedure and a commercial DNA extraction kit, is also presented. Application of this technology to species identification in industrial MBM was investigates as part of this study.
Mol Cell Probes 2001 Feb
PMID:Species-specific PCR for the identification of ovine, porcine and chicken species in meta and bone meal (MBM). 1128 33


<< Previous 1 2 3 4 5 6 7 8 9 10 Next >>