Non-radioactive labeling and detection of nucleic acids. I. A novel DNA labeling and detection system based on digoxigenin: anti-digoxigenin ELISA principle (digoxigenin system).

A novel highly sensitive non-radioactive DNA labeling and detection system based on the ELISA principle has been developed. DNA is modified with the cardenolide-hapten digoxigenin by enzymatic incorporation of digoxigenin-labeled deoxyuridine-triphosphate with Klenow enzyme. Digoxigenin is linked to dUTP via an 11-atom linear spacer (Dig-[11]-dUTP).

Attempted transmission of Ehrlichia risticii (Rickettsiaceae) with Stomoxys calcitrans (Diptera: Muscidae).

Experimental transmission of Ehrlichia risticii, the causal agent of Potomac horse fever, was attempted with adult stable flies, Stomoxys calcitrans, (L.) using two feeding schedules. In schedule A, a set of 140 flies was allowed to feed once on an experimentally infected donor pony and once 24 h later on a recipient pony.

RNA editing in transcripts of the mitochondrial genes of the insect trypanosome Crithidia fasciculata.

With the aid of cDNA and RNA sequence analysis, we have determined to what extent transcripts of mitochondrial maxicircle genes of the insect trypanosome Crithidia fasciculata are altered by RNA editing, a novel mechanism of gene expression which operates via the insertion and deletion of uridine residues. Editing of cytochrome c oxidase (cox) subunit II and III transcripts and of maxicircle unidentified reading frame (MURF) 2 RNA is limited to a small section and results in the creation of a potential AUG translational initiation codon (coxIII, MURF2) or the removal of a frameshift (coxII). No differences with the genomic sequences were observed in the remainder of these RNAs.

[Developmental disorders in young children with Duchenne's muscular dystrophy: implications for diagnosis and treatment].

The article describes two toddlers having Duchenne muscular dystrophy. In addition to motor problems they also exhibit attention disorders, deficits in short-term memory, speech and language disorders, and poor achievement motivation. Similar problems are reported in older Duchenne children.

Extracardiac surgical complications in heart transplant recipients.

As the population of patients undergoing orthotopic heart transplantation increases, more patients are likely to develop surgical complications unrelated to the transplant procedure. This article reviews 38 extracardiac surgical complications sustained in 18 of 48 patients undergoing orthotopic heart transplantation at our institution over a 4-year period. Twenty-seven complications (71.

Direct and sensitive detection of a pathogenic protozoan, Toxoplasma gondii, by polymerase chain reaction.

We applied the polymerase chain reaction to detection of the pathogenic protozoan Toxoplasma gondii based on our identification of a 35-fold-repetitive gene (the B1 gene) as a target. Using this procedure, we were able to amplify and detect the DNA of a single organism directly from a crude cell lysate. This level of sensitivity also allowed us to detect the B1 gene from purified DNA samples containing as few as 10 parasites in the presence of 100,000 human leukocytes.

Lack of selective developmental toxicity of three butanol isomers administered by inhalation to rats.

As part of an ongoing study of the developmental toxicology of industrial alcohols, this report presents the results of the teratology assessments of 1-butanol, 2-butanol, and t-butanol administered by inhalation to rats. Groups of approximately 15 Sprague-Dawley rats were exposed at 8000, 6000, 3500, or 0 ppm 1-butanol, 7000, 5000, 3500, or 0 ppm 2-butanol, or 5000, 3500, 2000, or 0 ppm t-butanol for 7 hr/day on Gestation Days 1-19 (sperm = 0). In each case, the highest concentration was selected to produce maternal toxicity.

Cloning of cDNAs encoding a 28 kilodalton antigen of Toxoplasma gondii.

By screening cDNA libraries in lambda gt11 with antibodies raised against the previously described protective F3G3 antigen of Toxoplasma gondii, and subsequently screening with nucleic acid probes, we have isolated cDNA clones that encode a 28 kDa antigen of T. gondii that is likely one of the two antigenic components of the F3G3 antigen. The gene apparently exists as a single copy in the tachyzoite haploid genome of the three strains of T.

Behavioral teratology investigation of 1-propanol administered by inhalation to rats.

Due to their structural similarity to ethanol, a human teratogen, and their widespread use in industry, a series of industrial alcohols are being investigated for developmental toxicity. This paper presents the results of exposures to 7000 ppm 1-propanol, which is minimally toxic to maternal animals and produces a low incidence of teratogenicity, and to 3500 ppm 1-propanol, which is not toxic to maternal rats and produces no teratogenicity. Propanol vapors or filtered air was administered for 7 hr/day to 15 pregnant Sprague-Dawley rats throughout gestation or to 18 male rats daily for 6 weeks.

Results of mechanical circulatory assistance before heart transplantation.

Between February 1984 and December 1987, 63 patients were accepted as candidates for heart transplantation. Eighteen patients (29%) required some form of mechanical circulatory support before transplantation; eight patients received an intraaortic balloon pump, five patients had left ventricular assist devices, two patients received biventricular assist devices, and in three patients the total artificial heart was implanted. Fourteen of the 18 patients underwent transplantation with seven longterm survivors.

Field and laboratory efficacy studies of erythrosin B for Musca domestica (Diptera: Muscidae) and Drosophila robusta (Diptera: Drosophilidae) control.

A 1.0% liquid bait formulation of erythrosin B was tested for house fly, Musca domestica L., control in one room of an environmentally controlled caged-layer poultry facility.

Molecular analysis of the gene encoding the major surface antigen of Toxoplasma gondii.

The complete sequence of P30, the major surface Ag of the protozoan parasite, Toxoplasma gondii, has been deduced through the cloning and analysis of its gene. Using polyclonal serum specific for P30, we have isolated a P30 cDNA clone from a lambda gt11 cDNA expression library derived from tachyzoites of T. gondii (RH strain).

Transcripts from the frameshifted MURF3 gene from Crithidia fasciculata are edited by U insertion at multiple sites.

In trypanosome mitochondria an RNA editing process is operative, which co- or post-transcriptionally alters the nucleotide sequence of transcripts by insertion and/or deletion of U residues at specific sites. To increase our understanding of the mechanism of this process we have compared the nucleotide sequence of the frameshifted mitochondrial MURF3 gene from Crithidia fasciculata to that of a large number of MURF3 cDNAs. We found cDNAs derived from transcripts edited at two different sites in the protein coding sequence: (i) at the frameshift position five extra U residues connect the two reading frames and (ii) at the 5' terminus 22 inserted Us shift a putative initiator codon out of phase.

Comparative analysis of precipitating antibodies in White Rock and Fayoumi hens injected with bovine serum albumin or crude mite extract with resulting effects on northern fowl mite, Ornithonyssus sylviarum (Acari: Macronyssidae) population densities.

Precipitating antibody concentration responses to crude northern fowl mite extract (CME) and bovine serum albumin (BSA) injections were compared in White Rock and Fayoumi hens with two-dimensional immunoelectrophoresis and rocket electrophoresis. The effect of CME injections on northern fowl mite population development was also determined. White Rock and Fayoumi hens developed similar antibody concentrations in response to intramuscular injections of BSA according to serum samples analyzed with two-dimensional immunoelectrophoresis.

Delayed reversal of impaired vasodilation in congestive heart failure after heart transplantation.

The effects of changes in central cardiovascular function on peripheral vasodilation were investigated. Strain gauge plethysmography was used to measure the maximal blood flow response following release of forearm arterial occlusion and the peak reactive hyperemic blood flow response (ml/min.100 ml) before and twice after orthotopic heart transplantation in 10 subjects with severe congestive heart failure.

Teratogenicity of n-propanol and isopropanol administered at high inhalation concentrations to rats.

As part of a teratological evaluation of several alcohols, 10,000, 7000 and 3500 ppm n-propanol or isopropanol were administered by inhalation to groups of 15 pregnant Sprague-Dawley rats for 7 hr/day on gestation days 1-19. The dams were killed on day 20. Half of the foetuses were examined for skeletal defects and the others for visceral defects using the Wilson technique.

Interlaboratory studies with the Chinese hamster V79 cell metabolic cooperation assay to detect tumor-promoting agents.

Three laboratories participated in an interlaboratory study to evaluate the usefulness of the Chinese hamster V79 cell metabolic cooperation assay to predict the tumor-promoting activity of selected chemicals. Twenty-three chemicals of different chemical structures (phorbol esters, barbiturates, phenols, artificial sweeteners, alkanes, and peroxides) were chosen for testing based on in vivo promotion activities, as reported in the literature. Assay protocols and materials were standardized, and the chemicals were coded to facilitate unbiased evaluation.

Neurochemical, but not behavioral, deviations in the offspring of rats following prenatal or paternal inhalation exposure to ethanol.

In addition to its widespread social use, ethanol is used extensively as an industrial solvent. Inhalation exposures to ethanol which produce narcosis in maternal rats are not teratogenic. The present study sought to extend the previous research by including offspring from paternal exposures, and testing for behavioral disorders in the offspring following maternal or paternal exposures.

Atrial natriuretic peptide in recipients of human orthotopic heart transplants.

Atrial natriuretic peptide (ANP) has been proposed to play a role in the maintenance of fluid and electrolyte homeostasis. ANP can be secreted in response to atrial stretch, and right atrial pressure has been proposed to control its secretion in vivo. This study was designed to characterize plasma immunoreactive (IR)-ANP in heart transplant recipients and to determine if IR-ANP levels in these patients correlate with cardiac filling pressures.

The nucleotide sequence of a 3.2 kb segment of mitochondrial maxicircle DNA from Crithidia fasciculata containing the gene for cytochrome oxidase subunit III, the N-terminal part of the apocytochrome b gene and a possible frameshift gene; further evidence for the use of unusual initiator triplets in trypanosome mitochondria.

A 3.2 kb segment of the maxicircle of Crithidia fasciculata mitochondrial (mt) DNA contains the gene for cytochrome oxidase subunit III (coxIII), the N-terminal portion of the gene for apocytochrome b (cytb) and two partially overlapping Unassigned Reading Frames (C.URF2/1).

Recommendations for the statistical analysis of Chernoff/Kavlock test data.

The statistical analyses of the data related to the teratogenic testing of 60 compounds utilizing the Chernoff/Kavlock test are the basis of this paper. The hypotheses chosen to be tested and the statistical tests utilized to test those hypotheses are discussed. The topics explored are the use of comparisonwise error rate versus experimentwise error rate, the use of nonparametric tests versus more "conventional" parametric tests and the use of "historical data" in assessing and interpreting group difference.

Evaluation of 60 chemicals in a preliminary developmental toxicity test.

The number of chemicals in commerce which have not been evaluated for potential developmental toxicity is large. Because of the time and expense required by conventional developmental toxicity tests, an abbreviated assay is needed that will preliminarily evaluate otherwise untested chemicals to help prioritize them for conventional testing. A proposed short-term in vivo assay has been used in a series of studies in which a total of 60 chemicals were tested.

Major transcript of the frameshifted coxII gene from trypanosome mitochondria contains four nucleotides that are not encoded in the DNA.

The mitochondrial cytochrome oxidase (cox) subunit II gene from trypanosomes contains a frameshift at amino acid 170. This gene is highly conserved in different trypanosome species, suggesting that it is functional. Sequence determination of coxII transcripts of T.