30 results on '"Matteson KJ"'
Search Results
2. Methemoglobin reduction in red cells: effect of a high oxygen affinity hemoglobin
- Author
-
Taketa, F, primary, Matteson, KJ, additional, Chen, JY, additional, and Libnoch, JA, additional
- Published
- 1980
- Full Text
- View/download PDF
3. Bioelectronic sensor technology for detection of cystic fibrosis and hereditary hemochromatosis mutations.
- Author
-
Bernacki SH, Farkas DH, Shi W, Chan V, Liu Y, Beck JC, Bailey KS, Pratt VM, Monaghan KG, Matteson KJ, Schaefer FV, Friez M, Shrimpton AE, and Stenzel TT
- Published
- 2003
- Full Text
- View/download PDF
4. Prevalence of urovirulence genes cnf, hlyD, sfa/foc, and papGIII in fecal Escherichia coli from healthy dogs and their owners.
- Author
-
Stenske KA, Bemis DA, Gillespie BE, Oliver SP, Draughon FA, Matteson KJ, and Bartges JW
- Subjects
- Animals, Dogs, Female, Humans, Virulence, Bacterial Toxins genetics, Escherichia coli genetics, Escherichia coli pathogenicity, Escherichia coli Proteins genetics, Feces microbiology, Hemolysin Proteins genetics, Membrane Transport Proteins genetics
- Abstract
Objective: To determine the prevalence of 4 urovirulence genes in fecal Escherichia coli isolates from healthy dogs and their owners and to determine whether detection of E coli strains with these genes was associated with a history of urinary tract infection (UTI)., Sample Population: 61 healthy dog-owner pairs and 30 healthy non-dog owners., Procedures: A fecal specimen was obtained from each participant, and 3 colonies of E coli were isolated from each specimen. A multiplex PCR assay was used to detect 4 genes encoding virulence factors: cytotoxic necrotizing factor (cnf), hemolysin (hlyD), s-fimbrial and F1C fimbriae adhesin (sfa/foc), and pilus associated with pyelonephritis G allele III (papGIII). Human participants completed a questionnaire to provide general information and any history of UTI for themselves and, when applicable, their dog., Results: 26% (16/61) of dogs, 18% (11/61) of owners, and 20% (6/30) of non-dog owners had positive test results for >or= 1 E coli virulence gene. One or more genes were identified in fecal E coli isolates of both dog and owner in 2% (1/61) of households. There was no difference in the detection of any virulence factor between dog-owner pairs. Female owner history of UTI was associated with detection of each virulence factor in E coli strains isolated from their dogs' feces., Conclusions and Clinical Relevance: Dogs and humans harbored fecal E coli strains possessing the genes cnf, hlyD, sfa/foc, and papGIII that encode urovirulence factors. It was rare for both dog and owner to have fecal E coli strains with these virulence genes.
- Published
- 2009
- Full Text
- View/download PDF
5. Comparison of clonal relatedness and antimicrobial susceptibility of fecal Escherichia coli from healthy dogs and their owners.
- Author
-
Stenske KA, Bemis DA, Gillespie BE, D'Souza DH, Oliver SP, Draughon FA, Matteson KJ, and Bartges JW
- Subjects
- Animals, Electrophoresis, Gel, Pulsed-Field, Escherichia coli isolation & purification, Hand Disinfection, Human-Animal Bond, Humans, Microbial Sensitivity Tests, Reference Values, Surveys and Questionnaires, Anti-Bacterial Agents pharmacology, Dogs microbiology, Escherichia coli drug effects, Feces microbiology
- Abstract
Objective: To determine prevalence of within-household sharing of fecal Escherichia coli between dogs and their owners on the basis of pulsed-field gel electrophoresis (PFGE), compare antimicrobial susceptibility between isolates from dogs and their owners, and evaluate epidemiologic features of cross-species sharing by use of a questionnaire., Sample Population: 61 healthy dog-owner pairs and 30 healthy control humans., Procedures: 3 fecal E coli colonies were isolated from each participant; PFGE profiles were used to establish relatedness among bacterial isolates. Susceptibility to 17 antimicrobials was determined via disk diffusion. A questionnaire was used to evaluate signalment, previous antimicrobial therapy, hygiene, and relationship with dog., Results: A wide array of PFGE profiles was observed in E coli isolates from all participants. Within-household sharing occurred with 9.8% prevalence, and across-household sharing occurred with 0.3% prevalence. No behaviors were associated with increased clonal sharing between dog and owner. No differences were found in susceptibility results between dog-owner pairs. Control isolates were more likely than canine isolates to be resistant to ampicillin and trimethoprim-sulfamethoxazole. Owners and control humans carried more multdrug-resistant E coli than did dogs., Conclusions and Clinical Relevance: Within-household sharing of E coli was detected more commonly than across-household sharing, but both direct contact and environmental reservoirs may be routes of cross-species sharing of bacteria and genes for resistance. Cross-species bacterial sharing is a potential public health concern, and good hygiene is recommended.
- Published
- 2009
- Full Text
- View/download PDF
6. Maternal antibrain antibodies in autism.
- Author
-
Zimmerman AW, Connors SL, Matteson KJ, Lee LC, Singer HS, Castaneda JA, and Pearce DA
- Subjects
- Adolescent, Adult, Animals, Autoantibodies, Case-Control Studies, Child, Child, Preschool, Female, Humans, Male, Maternal-Fetal Exchange, Middle Aged, Pregnancy, Antibody Formation, Autistic Disorder immunology, Brain immunology, Nerve Tissue Proteins immunology
- Abstract
Autism is a neurodevelopmental disorder of prenatal onset that is behaviorally defined. There is increasing evidence for systemic and neuroimmune mechanisms in children with autism. Although genetic factors are important, atypical prenatal maternal immune responses may also be linked to the pathogenesis of autism. We tested serum reactivity in 11 mothers and their autistic children, maternal controls, and several groups of control children, to prenatal, postnatal, and adult rat brain proteins, by immunoblotting. Similar patterns of reactivity to prenatal (gestational day 18), but not postnatal (day 8) or adult rat brain proteins were identified in autistic children, their mothers, and children with other neurodevelopmental disorders, and differed from mothers of normal children, normal siblings of children with autism and normal child controls. Specific patterns of antibody reactivity were present in sera from the autism mothers, from 2 to 18 years after the birth of their affected children and were unrelated to birth order. Immunoblotting using specific antigens for myelin basic protein (MBP) and glial acidic fibrillary protein (GFAP) suggests that these proteins were not targets of the maternal antibodies. The identification of specific serum antibodies in mothers of children with autism that recognize prenatally expressed brain antigens suggests that these autoantibodies could cross the placenta and alter fetal brain development.
- Published
- 2007
- Full Text
- View/download PDF
7. HLA-DR4 in families with autism.
- Author
-
Lee LC, Zachary AA, Leffell MS, Newschaffer CJ, Matteson KJ, Tyler JD, and Zimmerman AW
- Subjects
- Adult, Autistic Disorder immunology, Case-Control Studies, Catchment Area, Health, Child, Fathers, Female, Gene Frequency, Histocompatibility Testing, Humans, Male, Mothers, United States, White People genetics, Autistic Disorder genetics, HLA-DR4 Antigen genetics
- Abstract
Autoimmune disorders are observed with increased frequency among parents of individuals with autism, particularly mothers. Because there is evidence supporting an association between autoimmune disorders and specific alleles of the human leukocyte antigen (HLA) system, we examined HLA types and subtypes in families with autism. Two groups were studied: 16 families selected from a geographically defined area in eastern Tennessee have males with a diagnosis of autism; and 33 families selected across all regions in the United States have multiple males having autism diagnosis. The HLA-DR4 frequencies of mothers, fathers, and children in these two groups were compared with a reference series of 475 normal, unrelated Caucasians. Results of HLA typing indicated that mothers and their sons in the geographically defined group had a significantly higher frequency of DR4 than normal control subjects (odds ratio = 5.54, 95% confidence interval = 1.74-18.67 and odds ratio = 4.20, 95% confidence interval = 1.37-13.27, respectively). No significant difference in the distribution of HLA alleles was evident between the United States-all region group and control subjects. Findings of this study are consistent with a hypothesis that prenatal maternal-fetal immune interaction can affect fetal brain development in a population residing in a geographically defined region. Such immune interactions may involve HLA and related genes in both genetic and epigenetic mechanisms during pregnancy.
- Published
- 2006
- Full Text
- View/download PDF
8. Plasma serotonin in autism.
- Author
-
Connors SL, Matteson KJ, Sega GA, Lozzio CB, Carroll RC, and Zimmerman AW
- Subjects
- Adolescent, Adult, Autistic Disorder genetics, Case-Control Studies, Child, Child, Preschool, Fathers, Female, Humans, Male, Middle Aged, Mothers, Pilot Projects, Siblings, Autistic Disorder blood, Serotonin blood, Tryptophan blood
- Abstract
Serotonin is necessary for normal fetal brain development. Administration of serotonin inhibitors to pregnant rats results in offspring with abnormal behaviors, brain morphology, and serotonin receptor numbers. Low maternal plasma serotonin may contribute to abnormal brain development in autism. In this study, plasma serotonin levels in autism mothers and control mothers of typically developing children were compared, and plasma serotonin levels in children with autism (n = 17) and their family members were measured. Plasma serotonin levels in autism mothers were significantly lower than in mothers of normal children (P = 0.002). Plasma serotonin levels correlated between autism mothers and their children, but differed between autistic children and their fathers (P = 0.028) and siblings (P = 0.063). Low maternal plasma serotonin may be a risk factor for autism through effects on fetal brain development.
- Published
- 2006
- Full Text
- View/download PDF
9. Genetically characterized positive control cell lines derived from residual clinical blood samples.
- Author
-
Bernacki SH, Beck JC, Stankovic AK, Williams LO, Amos J, Snow-Bailey K, Farkas DH, Friez MJ, Hantash FM, Matteson KJ, Monaghan KG, Muralidharan K, Pratt VM, Prior TW, Richie KL, Levin BC, Rohlfs EM, Schaefer FV, Shrimpton AE, Spector EB, Stolle CA, Strom CM, Thibodeau SN, Cole EC, Goodman BK, and Stenzel TT
- Subjects
- Genetic Diseases, Inborn diagnosis, Humans, Laboratories, Molecular Biology, Mutation, Point Mutation, Sequence Deletion, Blood Specimen Collection, Cell Line, Transformed, Genetic Testing methods, Herpesvirus 4, Human, Lymphocytes cytology
- Abstract
Background: Positive control materials for clinical diagnostic molecular genetic testing are in critically short supply. High-quality DNA that closely resembles DNA isolated from patient specimens can be obtained from Epstein-Barr virus (EBV)-transformed peripheral blood lymphocyte cell lines. Here we report the development of a process to (a) recover residual blood samples with clinically important mutations detected during routine medical care, (b) select samples likely to provide viable lymphocytes for EBV transformation, (c) establish stable cell lines and confirm the reported mutation(s), and (d) validate the cell lines for use as positive controls in clinical molecular genetic testing applications., Methods: A network of 32 genetic testing laboratories was established to obtain anonymous, residual clinical samples for transformation and to validate resulting cell lines for use as positive controls. Three panel meetings with experts in molecular genetic testing were held to evaluate results and formulate a process that could function in the context of current common practices in molecular diagnostic testing., Results: Thirteen laboratories submitted a total of 113 residual clinical blood samples with mutations for 14 genetic disorders. Forty-one EBV-transformed cell lines were established. Thirty-five individual point and deletion mutations were shown to be stable after 20 population doublings in culture. Thirty-three cell lines were characterized for specific mutations and validated for use as positive controls in clinical diagnostic applications., Conclusions: A process for producing and validating positive control cell lines from residual clinical blood samples has been developed. Sustainable implementation of the process could help alleviate the current shortage of positive control materials.
- Published
- 2005
- Full Text
- View/download PDF
10. New directions in cytogenetic and molecular testing of the neonate.
- Author
-
Anderson IJ and Matteson KJ
- Subjects
- Humans, In Situ Hybridization, Fluorescence methods, Infant, Newborn, Karyotyping methods, Nucleic Acid Hybridization methods, Cytogenetic Analysis methods, Genetic Diseases, Inborn diagnosis, Genetic Diseases, Inborn genetics, Molecular Diagnostic Techniques methods
- Abstract
The development of new diagnostic, and hence therapeutic possibilities, has brought the realization that genetic disease is now an integral part of medical practice. Advances in cytogenetic and molecular testing have drastically improved the ability to diagnose with certainty many previously unrecognized conditions. However, this advance in technology does not come without new questions. New tests are not always the most cost effective ones, some have significant diagnostic limitations, and others raise valid ethical issues surrounding the testing of minors. A working understanding of new advances in genetic diagnosis as well as their inherent limitations is crucial for the contemporary practitioner.
- Published
- 2005
- Full Text
- View/download PDF
11. Effects of processing techniques on the forensic DNA analysis of human skeletal remains.
- Author
-
Arismendi JL, Baker LE, and Matteson KJ
- Subjects
- Female, Hot Temperature adverse effects, Humans, Male, Polymerase Chain Reaction, Ribs metabolism, Tandem Repeat Sequences, DNA isolation & purification, DNA Fingerprinting methods, Forensic Anthropology methods
- Abstract
Human remains processed by forensic anthropologists may potentially be used for genetic analysis. Therefore, the condition of the deoxyribonucleic acid (DNA) in processed remains may become an issue for future analysis. Processing techniques employed by anthropologists are highly variable and scanning electron microscopy reveals significant alterations to the bone surface depending upon the technique used. Such damage to the bone indicates differences may exist in quality and quantity of DNA extracted. This study assessed how five processing procedures used by major forensic anthropology laboratories around the country affects the amounts of DNA extracted from human rib bones and the subsequent DNA analysis. The DNA was analyzed using the short tandem repeat (STR) locus CSF1PO and amelogenin. The findings indicate processing procedures used by forensic anthropologists do not adversely affect DNA analysis but prolonged exposure to heat during processing may decrease the yield of information from the DNA.
- Published
- 2004
12. Establishment of stably EBV-transformed cell lines from residual clinical blood samples for use in performance evaluation and quality assurance in molecular genetic testing.
- Author
-
Bernacki SH, Stankovic AK, Williams LO, Beck JC, Herndon JE, Snow-Bailey K, Prior TW, Matteson KJ, Wasserman LM, Cole EC, and Stenzel TT
- Subjects
- Adult, Aging, Anticoagulants pharmacology, Cell Line, Transformed, Evaluation Studies as Topic, Female, Genetic Testing methods, Humans, Lymphocytes drug effects, Lymphocytes metabolism, Male, Middle Aged, Molecular Biology methods, Quality Control, Sex Characteristics, Temperature, Time Factors, Blood Specimen Collection, Cell Culture Techniques methods, Genetic Testing standards, Herpesvirus 4, Human physiology, Lymphocytes cytology, Lymphocytes virology, Molecular Biology standards
- Abstract
Positive control materials for clinical molecular genetic testing applications are currently in critically short supply or non-existent for many genetically based diseases of public health importance. Here we demonstrate that anonymous, residual, clinical blood samples are potential sources of viable lymphocytes for establishing Epstein-Barr virus (EBV)-transformed blood lymphocyte cell lines. We attempted to transform 34 residual blood samples, and analyzed transformation success with respect to sample age, anticoagulant, storage temperature, volume, hemolysis, and patient age and sex. In univariate analysis, sample age was significantly associated with transformation success (P = 0.002). The success rate was 67% (6 of 9) for samples 1 to 7 days old, 38% (3 of 8) for samples 8 to 14 days old and 0% for samples 15 to 21 (0 of 11) days old. When we controlled for sample age in multivariate logistic regression, anticoagulant and storage temperature approached significance (P = 0.070 and 0.087, respectively; samples in acid citrate dextrose (ACD) and refrigerated samples were more likely to transform). Based on these findings, we suggest that samples collected in either ACD or ethylene diamine tetraacetic acid, and up to 14 days old (refrigerated) or 7 days old (stored ambient), are reasonable candidates for EBV transformation. The transformation rate for samples that met these criteria was 63% (10 of 16). Implementation of this process could help alleviate the shortage of positive control materials for clinical molecular genetic testing.
- Published
- 2003
- Full Text
- View/download PDF
13. A silica-based mitochondrial DNA extraction method applied to forensic hair shafts and teeth.
- Author
-
Baker LE, McCormick WF, and Matteson KJ
- Subjects
- Adult, Aged, Aged, 80 and over, Autopsy, DNA Fingerprinting, Female, Hair chemistry, Humans, Male, Middle Aged, Polymerase Chain Reaction, Silicon Dioxide, Tooth chemistry, DNA, Mitochondrial genetics, DNA, Mitochondrial isolation & purification, Forensic Medicine methods
- Abstract
The purpose of this study is to evaluate the applicability of a nonorganic DNA extraction method for use in the analysis of environmentally compromised forensic hair shaft and tooth samples. The condition of the samples included cases of water decomposition, severe incineration, and varying stages of putrefaction. Enzymatic amplification and manual sequencing of the first segment of the mitochondrial hypervariable region were performed successfully on each of the 20 autopsied individuals. The results indicate that the silica-based extraction method produces mtDNA suitable for genetic identification from forensic samples including hair shafts and teeth.
- Published
- 2001
14. Laser desorption mass spectrometry for point mutation detection.
- Author
-
Taranenko NI, Matteson KJ, Chung CN, Zhu YF, Chang LY, Allman SL, Haff L, Martin SA, and Chen CH
- Subjects
- Cell Line, DNA Probes, Humans, Polymerase Chain Reaction, Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization, Cystic Fibrosis Transmembrane Conductance Regulator genetics, Point Mutation
- Abstract
A point mutation can be associated with the pathogenesis of inherited or acquired diseases. Laser desorption mass spectrometry coupled with allele specific polymerase chain reaction (PCR) was first used for point mutation detection. G551D is one of several mutations of the cystic fibrosis transmembrane conductance regulator (CFTR) gene present in 1-3% of the mutant CFTR alleles in most European populations. In this work, two different approaches were pursued to detect G551D point mutation in the cystic fibrosis gene. The strategy is to amplify the desired region of DNA template by PCR using two primers that overlap one base at the site of the point mutation and which vary in size. If the two primers based on the normal sequence match the target DNA sequence, a normal PCR product will be produced. However, if the alternately sized primers that match the mutant sequence recognize the target DNA, an abnormal PCR product will be produced. Thus, the mass spectrometer can be used to identify patients that are homozygous normal, heterozygous for a mutation or homozygous abnormal at a mutation site. Another approach to identify similar mutations is the use of sequence specific restriction enzymes which respond to changes in the DNA sequence. Mass spectrometry is used to detect the length of the restriction fragments generated by digestion of a PCR generated target fragment.
- Published
- 1996
- Full Text
- View/download PDF
15. Detection of delta F508 mutation of the cystic fibrosis gene by matrix-assisted laser desorption/ionization mass spectrometry.
- Author
-
Ch'ang LY, Tang K, Schell M, Ringelberg C, Matteson KJ, Allman SL, and Chen CH
- Subjects
- Base Sequence, Humans, Lasers, Mass Spectrometry, Molecular Sequence Data, Oligonucleotide Probes, Polymerase Chain Reaction, Cystic Fibrosis genetics, DNA analysis, Mutation
- Abstract
The most common mutation of the cystic fibrosis gene is characterized by the deletion of three nucleotides that code phenylalanine in the 508 position of the cystic fibrosis transmembrane conductance regulator. We report the first measurements by matrix-assisted laser desorption/ionization (MALDI) time-of-flight mass spectrometry for the delta F508 mutation in cystic fibrosis carriers and patients. Furthermore, in a blind test, results from the normal and delta F508 mutant alleles in 30 clinical samples based on MALDI mass spectrometry and on conventional gel analysis of the DNA were in total agreement. These results demonstrate the utility of MALDI mass spectrometry in the molecular diagnosis of mutant alleles and point to its potential use for ultra-fast detection in large-scale screening of DNA mutations.
- Published
- 1995
- Full Text
- View/download PDF
16. HPLC analysis of amino acids in inborn errors of metabolism.
- Author
-
Matteson KJ
- Subjects
- Amino Acid Metabolism, Inborn Errors blood, Amino Acid Metabolism, Inborn Errors urine, Amino Acids urine, Chromatography, High Pressure Liquid methods, Diagnosis, Differential, Humans, Indicators and Reagents, Reference Values, Amino Acid Metabolism, Inborn Errors diagnosis, Amino Acids blood
- Abstract
Analysis of amino acids in blood or urine is a valuable diagnostic tool in cases of suspected metabolic disorders. The presence of a characteristic pattern of elevated amino acids is very useful in the diagnosis of these rare disorders. The detection of an apparently normal pattern of amino acids is also helpful to the clinician since it will eliminate many inborn errors of metabolism from the list of potential disorders. Methodologies for amino acid analysis in physiological fluids range from the very simple thin layer chromatography to automated low pressure or high pressure chromatography. Low pressure chromatography using a Beckman analyzer or similar instrument is the most common methodology for physiological amino acid analysis. Chromatography (HPLC) systems for amino acid analysis of proteins are available that can be modified for use with physiological samples. Waters makes a system called Picotag(TM) and Applied Biosystems makes an automated analyzer. These HPLC systems have some advantages over LPLC systems, including lower equipment cost, less caustic buffer systems and improved separation of certain amino acids.
- Published
- 1995
17. Use of a molecular genetic approach to diagnosing the fragile X genotype.
- Author
-
Potter NT, Lozzio CB, Anderson IJ, Bowlin ES, and Matteson KJ
- Subjects
- Blotting, Southern, Cytogenetics, Female, Fragile X Syndrome genetics, Genetic Carrier Screening methods, Genetic Linkage, Genotype, Humans, Male, Pedigree, Fragile X Syndrome diagnosis
- Abstract
We report the direct molecular detection of the fragile X genotype in 111 individuals from 17 families with a total of 31 cases of fragile X syndrome. Comparison of our molecular data with our previous cytogenetic and linkage data from these same families indicates the effectiveness of the direct molecular analysis. We have been able to assign a genotype unambiguously in 100% of the persons tested, and in all cases the molecular data correlated with the cytogenetic or linkage findings or both. Two of the three families presented in this study represent inheritance of this gene through normal transmitting males, and the third is strongly suggestive of this mode of inheritance. Our data show that the direct molecular approach will be of great utility for confirmation of the diagnosis and for the detection of female carriers and normal transmitting males who are at high risk for having affected children or grandchildren.
- Published
- 1992
- Full Text
- View/download PDF
18. Regional evaluation of DNA diagnostic laboratories.
- Author
-
Matteson KJ, Barker PE, Kaplan GC, Mueller OT, Ostrer H, Phillips JA 3rd, and Schwartz C
- Subjects
- Evaluation Studies as Topic, DNA genetics, Diagnostic Services standards, Laboratories standards
- Published
- 1990
19. An approach to the molecular biology of congenital adrenal hyperplasia.
- Author
-
Chung BC, Matteson KJ, Morin JE, Mellon SH, and Miller WL
- Subjects
- Animals, Base Sequence, Cattle, Cloning, Molecular, Humans, Adrenal Hyperplasia, Congenital genetics, Cytochrome P-450 Enzyme System genetics, DNA genetics, DNA, Recombinant, Steroid 21-Hydroxylase genetics, Steroid Hydroxylases genetics
- Published
- 1985
- Full Text
- View/download PDF
20. A study of restriction fragment length polymorphisms at the human alpha-1-antitrypsin locus.
- Author
-
Matteson KJ, Ostrer H, Chakravarti A, Buetow KH, O'Brien WE, Beaudet AL, and Phillips JA
- Subjects
- Base Sequence, Cloning, Molecular, DNA genetics, DNA Restriction Enzymes, Electrophoresis, Agar Gel, Genetic Linkage, Genetic Markers, Humans, Nucleic Acid Hybridization, Chromosome Mapping, Polymorphism, Genetic, alpha 1-Antitrypsin genetics
- Abstract
A cloned cDNA for alpha-1-antitrypsin (alpha-1-AT) was selected from a human liver cDNA library. The identity of the clone was established by hybrid-selected translation and partial DNA sequencing. The cDNA was used as a probe to search for restriction site polymorphisms (RSPs) near the alpha-1-AT gene. Only two RSPs were found using 29 different restriction enzymes. Each of these polymorphisms resulted from the loss of a restriction site, one for EcoRI and the other for Taq I. The frequency of polymorphic restriction was calculated to be 1.1% to 2.6% of all sites tested, a figure lower than the 9.3% value observed for 12 RSPs in the human beta-globin gene cluster. Since the corresponding figure for detectable polymorphisms at the alpha-1-AT locus at the protein level is 12%, restriction enzymes are comparatively inefficient in detecting genetic variability. The basis of this inefficiency was studied by computing the nucleotide diversity from the RSP data. On the average, one in 500 to 1000 bases is polymorphic around the alpha-1-At locus. This value is comparable to that which we have calculated for the human beta-globin gene cluster and the human growth hormone gene cluster (both one in 500). These data demonstrate the limited usefulness of linked RSPs for genetic linkage studies at the alpha-1-AT locus.
- Published
- 1985
- Full Text
- View/download PDF
21. Inhibition of NADH-methemoglobin reductase by organic phosphates.
- Author
-
Matteson KJ and Taketa F
- Subjects
- 2,6-Dichloroindophenol metabolism, Ferricyanides metabolism, Humans, Inositol Phosphates pharmacology, Methemoglobin metabolism, Sulfhydryl Reagents pharmacology, Cytochrome-B(5) Reductase antagonists & inhibitors, NADH, NADPH Oxidoreductases antagonists & inhibitors, Organophosphorus Compounds pharmacology
- Abstract
The organic phosphate allosteric effectors of hemoglobin, inositol hexaphosphate, 2,3-diphosphoglycerate, and ATP, interact with NADH-methemoglobin reductase (NADH-diaphorase). Significant inhibitory effects on the enzyme were found when dichlorophenolindophenol, or ferricyanide were used as electron acceptors in place of methemoglobin. In contrast, apparent stimulation of enzyme activity was observed when adult human methemoglobin was used as the electroganic phosphate on the rate of reaction due to its interaction with the substrate methemoglobin to produce the favored T type of quaternary conformation. The inhibitory effect of inositol hexaphosphate on the enzyme is associated with a perturbation in the reactivity of essential sulfhydryl group(s) on the enzyme. It is suggested that the interaction of the organic phosphate with the enzyme as well as with the substrate is significant in determining the overall rate of methemoglobin reduction.
- Published
- 1979
- Full Text
- View/download PDF
22. Study of cholesterol side-chain cleavage (20,22 desmolase) deficiency causing congenital lipoid adrenal hyperplasia using bovine-sequence P450scc oligodeoxyribonucleotide probes.
- Author
-
Matteson KJ, Chung BC, Urdea MS, and Miller WL
- Subjects
- Adrenal Hyperplasia, Congenital complications, Amino Acid Sequence, Animals, Base Sequence, Cattle, DNA analysis, DNA biosynthesis, DNA Restriction Enzymes metabolism, Humans, Lipidoses complications, Nucleic Acid Hybridization, Oligonucleotides analysis, Poly A metabolism, RNA metabolism, RNA, Messenger metabolism, Repetitive Sequences, Nucleic Acid, Substrate Specificity, Adrenal Hyperplasia, Congenital enzymology, Cholesterol Side-Chain Cleavage Enzyme deficiency, Lipidoses enzymology, Oxidoreductases deficiency
- Abstract
Conversion of cholesterol to pregnenolone is mediated by the cholesterol side-chain cleavage (SCC) enzyme, P450scc. Deficient SCC activity causes congenital lipoid adrenal hyperplasia (also known as 20,22 desmolase deficiency), a potentially lethal defect in the synthesis of all steroid hormones. To probe for possible genetic defects causing this disease we synthesized four oligodeoxyribonucleotides containing 63 to 72 bases corresponding to portions of the bovine complementary DNA (cDNA) sequence for P450scc. The bovine oligonucleotides were labeled and used directly to probe Southern blots of normal human genomic DNA, revealing a pattern indicating there is a single P450scc gene in the human genome. Hybridization to Northern blots of normal human and bovine adrenal messenger RNA indicates that P450scc messenger RNA is about 2.0 kilobases long in both species. Hybridizations of the oligonucleotides to genomic DNA from three unrelated patients with SCC deficiency did not detect a deletion in the human P450scc gene. The bovine sequence oligonucleotides were then used to isolate a human P450scc cDNA clone. The isolated P450scc cDNA fragment contains 818 bases encoding 239 amino acids of the protein, the translation termination signal, and 98 bases of the 3' untranslated region. The sequence of this carboxy-terminal half of the human P450scc protein is 72% homologous with the bovine sequence and contains an additional amino acid not found in bovine P450scc; the human and bovine nucleotide sequences are 81% homologous. Repetition of the genomic DNA blotting studies with the cDNA probe gave the same results obtained with the bovine-sequence oligonucleotide probes, confirming that SCC deficiency is not due to a deletion in the regions of the P450scc hybridizing with the probes. Long, chemically synthesized heterologous sequence oligonucleotides containing unknown numbers of base mismatches with human sequences may thus be used to study human genes so that access to a cDNA is not necessary for such studies.
- Published
- 1986
- Full Text
- View/download PDF
23. Molecular cloning of DNA complementary to bovine adrenal P450scc mRNA.
- Author
-
Matteson KJ, Chung BC, and Miller WL
- Subjects
- Adrenal Glands metabolism, Animals, Cattle, Chemical Precipitation, Cloning, Molecular, Immunochemistry, Nucleic Acid Hybridization, Protein Biosynthesis, Cytochrome P-450 Enzyme System genetics, DNA, RNA, Messenger genetics
- Abstract
P450scc is the rate-limiting hormonally regulated enzyme that cleaves the cholesterol side chain. Translation of bovine adrenocortical mRNA and immunoprecipitation with rabbit anti-bovine P450scc indicates P450scc mRNA represents 1% of the total. DNA complementary to bovine adrenocortical mRNA was cloned in the PstI site of pBR322 by dC X dG tailing and high-efficiency transformation. A clone containing sequences complementary to P450scc mRNA was identified by hybrid-selected translation only when plasmid DNA was first purified by CsCl gradient centrifugation. As is often the case with hybrid-selected translation, the clone identified contains a small insert.
- Published
- 1984
- Full Text
- View/download PDF
24. Structure of a bovine gene for P-450c21 (steroid 21-hydroxylase) defines a novel cytochrome P-450 gene family.
- Author
-
Chung BC, Matteson KJ, and Miller WL
- Subjects
- Amino Acid Sequence, Animals, Cattle, Gene Expression Regulation, Genes, Glucocorticoids physiology, Sequence Homology, Nucleic Acid, Cytochrome P-450 Enzyme System genetics, Steroid 21-Hydroxylase genetics, Steroid Hydroxylases genetics
- Abstract
P-450c21, a cytochrome P-450 enzyme [steroid 21-monooxygenase (steroid 21-hydroxylase), EC 1.14.99.10], mediates the 21-hydroxylation of glucocorticoid and mineralocorticoid hormones in the adrenal gland. The complete sequence of a bovine P-450c21 gene shows it is 3447 base pairs long and contains 10 exons. The intron/exon organization and encoded amino acid sequence indicate that P-450c21 represents a unique family of genes in the P-450 gene superfamily. Primer extension and S1 nuclease protection experiments identified several cap sites for initiation of transcription; the principal cap site produces mRNA with a 5' untranslated region only 11 bases long. S1 nuclease protection experiments confirm that P-450c21 is actively expressed in the adrenal and the testis, an organ not known to secrete 21-hydroxylated steroids.
- Published
- 1986
- Full Text
- View/download PDF
25. Human cholesterol side-chain cleavage enzyme, P450scc: cDNA cloning, assignment of the gene to chromosome 15, and expression in the placenta.
- Author
-
Chung BC, Matteson KJ, Voutilainen R, Mohandas TK, and Miller WL
- Subjects
- Amino Acid Sequence, Base Sequence, Cloning, Molecular, Cyclic AMP physiology, DNA genetics, Gene Expression Regulation, Humans, Placenta physiology, Cholesterol Side-Chain Cleavage Enzyme genetics, Chromosomes, Human, Pair 15, Oxidoreductases genetics
- Abstract
Conversion of cholesterol to pregnenolone is mediated by P450scc [cholesterol, reduced-adrenal-ferrodoxin: oxygen oxidoreductase (side-chain-cleaving), EC 1.14.15.67]. RNA from several human adrenal samples was translated in vitro and immunoprecipitated with anti-bovine P450scc, indicating that P450scc mRNA represents about 0.5% of human adrenal mRNA in normal, hypertrophied, and malignant adrenals. A 1626-base-pair human adrenal P450scc cDNA was cloned in bacteriophage lambda gt10. Primer extension data indicated P450scc mRNA is about 1850 bases long and that all adrenal P450scc mRNA has the same 5' end. A full-length clone containing 1821 bases was obtained from a human testis cDNA library to yield the complete sequence. The encoded human preP450scc contains 521 amino acids with a molecular weight of 60189.65. The testis and adrenal sequences were identical; the human cDNA and amino acid sequences are 82% and 72% homologous, respectively, with the bovine sequences. P450scc cDNA was used to probe DNA from a panel of mouse-human somatic cell hybrids, showing that the single human P450scc gene lies on chromosome 15. The human P450scc gene is expressed in the placenta in early and midgestation; primary cultures of placental tissue indicate P450scc mRNA accumulates in response to cyclic AMP.
- Published
- 1986
- Full Text
- View/download PDF
26. Hormonal regulation of P450scc (20,22-desmolase) and P450c17 (17 alpha-hydroxylase/17,20-lyase) in cultured human granulosa cells.
- Author
-
Voutilainen R, Tapanainen J, Chung BC, Matteson KJ, and Miller WL
- Subjects
- Aldehyde-Lyases genetics, Bucladesine pharmacology, Butyrates pharmacology, Butyric Acid, Cells, Cultured, Cholesterol Side-Chain Cleavage Enzyme genetics, Chorionic Gonadotropin pharmacology, DNA, Female, Follicle Stimulating Hormone pharmacology, Humans, Progesterone metabolism, Prolactin pharmacology, RNA, Messenger metabolism, Steroid 17-alpha-Hydroxylase genetics, Aldehyde-Lyases biosynthesis, Cholesterol Side-Chain Cleavage Enzyme biosynthesis, Gene Expression Regulation drug effects, Granulosa Cells enzymology, Oxidoreductases biosynthesis, Steroid 17-alpha-Hydroxylase biosynthesis, Steroid Hydroxylases biosynthesis
- Abstract
Conversion of cholesterol to pregnenolone in man is mediated by a single enzyme, P450scc. To study possible regulation of the single P450scc gene in ovarian steroid synthesis, we incubated human granulosa cells with potential hormonal stimulators, measured P450scc mRNA accumulation by hybridization to 32P-labeled human P450scc cDNA, and compared the results to secretion of progesterone into the culture medium. Primary cultures of human granulosa cells were optimally responsive after 8-14 days of culture. Incubation with hCG (1.0-100 ng/ml), FSH (1.0-50 ng/ml), and (Bu)2cAMP (0.02-2.0 mM) increased P450scc mRNA accumulation and progesterone secretion in dose-dependent fashions. Maximal stimulation increased P450scc mRNA accumulation and progesterone secretion to 490% and 240% of control values, respectively, with hCG, to 166% and 168% with FSH, and to 495% and 380% with (Bu)2cAMP. PRL (to 100 ng/ml), ACTH (10(-6) M), and butyric acid (2 mM) had no significant effect on progesterone secretion or P450scc mRNA accumulation. These data indicate gonadotropin-specific stimulation of cAMP-mediated regulation of P450scc mRNA accumulation in human granulosa cells, presumably mediated by increased P450scc gene transcription. Ovarian estrogen synthesis may require both thecal and granulosa cells, although this two-cell theory of estrogen synthesis is unproven in man. To examine this theory, we probed the same blots used in the experiments described above with 32P-labeled human P450c17 cDNA (P450c17 is the single enzyme mediating both 17 alpha-hydroxylase and 17,20-lyase activities). Only miniscule amounts of P450c17 mRNA were found in the human granulosa cells, and the amounts did not increase in response to any of the above stimuli. These data strongly support the two-cell theory of human ovarian estrogen synthesis.
- Published
- 1986
- Full Text
- View/download PDF
27. P450XXI (steroid 21-hydroxylase) gene deletions are not found in family studies of congenital adrenal hyperplasia.
- Author
-
Matteson KJ, Phillips JA 3rd, Miller WL, Chung BC, Orlando PJ, Frisch H, Ferrandez A, and Burr IM
- Subjects
- Adrenal Hyperplasia, Congenital enzymology, Amino Acid Sequence, Base Sequence, Cytochrome P-450 Enzyme System genetics, DNA analysis, Haploidy, Humans, Mutation, Pedigree, Adrenal Hyperplasia, Congenital genetics, Chromosome Deletion, Steroid 21-Hydroxylase genetics, Steroid Hydroxylases genetics
- Abstract
Congenital adrenal hyperplasia (CAH) is a common genetic disorder due to defective 21-hydroxylation of steroid hormones. The human P450XXIA2 gene encodes cytochrome P450c21 [steroid 21-monooxygenase (steroid 21-hydroxylase), EC 1.14.99.10], which mediates 21-hydroxylation. The P450XXIA2 gene may be distinguished from the duplicated P450XXIA1 pseudogene by cleavage with the restriction endonuclease Taq I, with the XXIA2 gene characterized by a 3.7-kilobase (kb) fragment and the XXIA1 pseudogene characterized by a 3.2-kb fragment. Restriction endonuclease mapping by several laboratories has suggested that deletion of the P450XXIA2 gene occurs in about 25% of patients with CAH, as their genomic DNA lacks detectable 3.7-kb Taq I fragments. We have cloned human P450c21 cDNA and used it to study genomic DNA prepared from 51 persons in 10 families, each of which includes 2 or more persons with CAH. After Taq I digestion, apparent deletions are seen in 7 of the 20 alleles of the probands; using EcoRI, apparent deletions are seen in 9 of the 20 alleles. However, the apparently deleted alleles seen with Taq I do not coincide with those seen with EcoRI. Furthermore, studies with Bgl II, EcoRI, Kpn I, and Xba I yield normal patterns with at least two enzymes in all cases. Since all probands yielded normal patterns with at least two of the five enzymes used, we conclude that the P450XXIA2 gene "deletions" widely reported in CAH patients probably represent gene conversions, unequal crossovers, or polymorphisms rather than simple gene deletions.
- Published
- 1987
- Full Text
- View/download PDF
28. Cloning and characterization of the bovine gene for steroid 21-hydroxylase (P-450c21).
- Author
-
Chung BC, Matteson KJ, and Miller WL
- Subjects
- Amino Acid Sequence, Animals, Base Sequence, Cattle, Cloning, Molecular, Genes, Humans, Nucleic Acid Hybridization, Oligodeoxyribonucleotides chemical synthesis, Cytochrome P-450 Enzyme System genetics, Steroid 21-Hydroxylase genetics, Steroid Hydroxylases genetics
- Abstract
Steroid 21-hydroxylase activity is required for the synthesis of both cortisol and aldosterone. Using two manually synthesized oligonucleotide probes, we screened a bovine genomic DNA library and identified a phage, lambda E11, carrying the gene for the steroid 21-hydroxylase, P-450c21. The identity of lambda E11 was initially proven by initiating dideoxy sequencing from the two oligonucleotides directly on the full-length, uncleaved phage template. Hybridization of total bovine genomic DNA to lambda E11 restriction fragments indicates the presence of repetitive sequences in or near the P-450c21 gene. Northern blots indicate that the mature mRNA exists in two principal forms of about 2.2 and 2.4 kb in length. Southern blots indicate there are two copies of the gene in the bovine genome. Sequence analysis of a 1141-bp Eco RI fragment of the gene shows three complete exons and a portion of a fourth exon, correctly determining the amino acid sequence of 148 amino acids of this important enzyme. This cloned 1141-bp fragment cross-hybridizes to human genomic DNA, indicating it should be a useful probe for studying the human P-450c21 gene in patients having impaired 21-hydroxylase activity.
- Published
- 1985
- Full Text
- View/download PDF
29. Methemoglobin reductase: inactivation and protection against inactivation during disc gel electrophoresis.
- Author
-
Matteson KJ and Taketa F
- Subjects
- Cytochrome-B(5) Reductase antagonists & inhibitors, Dithiothreitol pharmacology, Electrophoresis, Disc, Globins pharmacology, Hemoglobins pharmacology, Serum Albumin, Bovine pharmacology, Cytochrome-B(5) Reductase blood, NADH, NADPH Oxidoreductases blood
- Abstract
Methemoglobin reductase was found to be inactivated during electrophoresis on ammonium persulfate polymerized polyacrylamide gels. Hemoglobin in various states of ligation offers protection from inactivation. Thiols such as dithiothreitol also offer protection but the effect due to hemoglobin is not provided by its sulfhydryl groups.
- Published
- 1979
- Full Text
- View/download PDF
30. Assignment of the gene for adrenal P450c17 (steroid 17 alpha-hydroxylase/17,20 lyase) to human chromosome 10.
- Author
-
Matteson KJ, Picado-Leonard J, Chung BC, Mohandas TK, and Miller WL
- Subjects
- Animals, DNA, Humans, Hybrid Cells, Mice, Nucleic Acid Hybridization, Adrenal Glands enzymology, Aldehyde-Lyases genetics, Chromosome Mapping, Chromosomes, Human, 6-12 and X, Cytochrome P-450 Enzyme System genetics, Steroid 17-alpha-Hydroxylase genetics, Steroid Hydroxylases genetics
- Abstract
P450c17 is the single enzyme mediating both 17 alpha-hydroxylase and 17,20 lyase activities. We identified several human P450c17 cDNA clones in a human adrenal cDNA library we constructed in lambda gt10. A short clone containing the 3'-terminal 650 bases of the full-length sequence was used to examine Southern blots of DNA from normal persons and from a panel of mouse/human somatic cell hybrid lines. The pattern of hybridization of this cDNA to normal human DNA cut with 8 restriction endonucleases suggests the human genome has two (or more) P450c17 genes. The pattern of hybridization to the somatic cell hybrid cell lines, each containing a limited, known number of human chromosomes, indicates the human adrenal P450c17 gene lies on chromosome 10. The chromosomal locations of the other P450c17 genes could not be determined.
- Published
- 1986
- Full Text
- View/download PDF
Catalog
Discovery Service for Jio Institute Digital Library
For full access to our library's resources, please sign in.