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Biomedical subjects

D T Dennis

Publications and source records attributed to D T Dennis.

At least 19 recordsLinked to original sources
PubMed1998

Emergence of Lyme disease in Hunterdon County, New Jersey, 1993: a case-control study of risk factors and evaluation of reporting patterns.

Reported cases of Lyme disease in Hunterdon County, New Jersey, increased almost 200% from 75 (67/100,000 population) in 1992 to 216 (193/100,000 population) in 1993. For evaluation of risk factors for Lyme disease and for determination of the cause of this increase, a case-control study was conducted, and the reporting practices of physicians' offices were evaluated. For cases reported in 1993, age and sex distribution, month of disease onset, and proportion of cases with erythema migrans rash were within expected limits. Analysis of age-matched case-control data showed that rural residence; clearing periresidential brush during spring and summer months; and the presence of rock walls, woods, deer, or a bird feeder on residential property were associated with incident Lyme disease. A review of physician reporting patterns suggested that the increase in reported cases in 1993 was due to improved reporting as well as to an increase in the numbers of patients diagnosed with Lyme disease. In addition, substantial underreporting of Lyme disease by physicians' offices was found.

Adolescent↗
PubMed1998

Test-treatment strategies for patients suspected of having Lyme disease: a cost-effectiveness analysis.

PURPOSE: To examine the cost-effectiveness of test-treatment strategies for patients suspected of having Lyme disease. DATA SOURCES: The medical literature was searched for information on outcomes and costs. Expert opinion was sought for information on utilities. STUDY SELECTION: Articles that described patient population, diagnostic criteria, dose and duration of therapy, and criteria for assessment of outcomes. DATA EXTRACTION: The decision analysis evaluated the following strategies: 1) no testing-no treatment; 2) testing with enzyme-linked immunosorbent assay (ELISA) followed by antibiotic treatment of patients with positive results; 3) two-step testing with ELISA followed by Western blot and antibiotic treatment for patients with positive results on either test; and 4) empirical antibiotic therapy. Three patient scenarios were considered: myalgic symptoms, rash resembling erythema migrans, and recurrent oligoarticular inflammatory arthritis. Results were calculated as costs per quality-adjusted life-year and were subjected to sensitivity analysis. Adjustment was made for the diagnostic value of common clinical features of Lyme disease. DATA SYNTHESIS: For myalgic symptoms without other features suggestive of Lyme disease, the no testing-no treatment strategy was most economically attractive (that is, had the most favorable cost-effectiveness ratio). For rash, empirical antibiotic therapy was less costly and more effective than other strategies. For oligoarticular arthritis with a history of rash and tick bite, two-step testing was associated with the lowest cost-effectiveness ratio. Testing with ELISA and empirical antibiotic therapy cost an additional $880,000 and $34,000 per quality-adjusted life-year, respectively. For oligoarticular arthritis with one or no other features suggestive of Lyme disease, two-step testing was most economically attractive. CONCLUSIONS: Neither testing nor antibiotic treatment is cost-effective if the pretest probability of Lyme disease is low. Empirical antibiotic therapy is recommended if the pretest probability is high, and two-step testing is recommended if the pretest probability is intermediate.

Anti-Bacterial Agents↗
PubMed1998

Altered growth of transgenic tobacco lacking leaf cytosolic pyruvate kinase.

Previously, we reported that transformation of tobacco (Nicotiana tabacum L.) with a vector containing a potato cytosolic pyruvate kinase (PKc) cDNA generated two plant lines specifically lacking leaf PKc (PKc-) as a result of co-suppression. PKc deficiency in these primary transformants did not appear to alter plant development, although root growth was not examined. Here we report a striking reduction in root growth of homozygous progeny of both PKc- lines throughout development under moderate (600 microE m-2 s-1) or low (100 microE m-2 s-1) light intensities. When both PKc- lines were cultivated under low light, shoot and flower development were also delayed and leaf indentations were apparent. Leaf PK activity in the transformants was significantly decreased at all time points examined, whereas root activities were unaffected. Polypeptides corresponding to PKc were undetectable on immunoblots of PKc- leaf extracts, except in 6-week-old low-light-grown PKc- plants, in which leaf PKc expression appeared to be greatly reduced. The metabolic implications of the kinetic characteristics of partially purified PKc from wild-type tobacco leaves are discussed. Overall, the results suggest that leaf PKc deficiency leads to a perturbation in source-sink relationships.

Cytosol↗
PubMed1997

Laboratory evaluation in the diagnosis of Lyme disease.

PURPOSE: To provide a qualitative evaluation of the predictive value of the laboratory diagnosis of Lyme disease and to use the resultant data to formulate guidelines for clinical diagnosis. DATA SOURCES: A MEDLINE search of English-language articles or articles with English-language abstracts published from 1982 to 1996. DATA EXTRACTION: Sensitivity, specificity, and likelihood ratios were calculated, and a random-effects model was used to combine the proportions from the eligible studies. Prespecified criteria were used to determine which studies were eligible for analysis. DATA SYNTHESIS: Laboratory testing in general is not clinically useful if the pretest probability of Lyme disease is less than 0.20 or greater than 0.80. When the pretest probability is 0.20 to 0.80, sequential testing with enzyme-linked immunosorbent assay and Western blot is the most accurate method for ruling in or ruling out the possibility of Lyme disease. CONCLUSIONS: Laboratory testing is recommended only in patients whose pretest probability of Lyme disease is 0.20 to 0.80. If the pretest probability is less than 0.20, testing will result in more false-positive results than true-positive results; a negative test result in this situation effectively rules out the disease.

Bacteriological Techniques↗
PubMed1996

Transit peptides play a major role in the preferential import of proteins into leucoplasts and chloroplasts.

The in vitro import characteristics of six different precursors of plastid proteins were assessed to determine differences in the protein import pathways of leucoplasts and chloroplasts. Five of these precursor proteins are destined to different subchloroplast sites, and one is a leucoplast stromal precursor protein. The results indicate that some of these precursors can be imported equally into both plastid types and others preferentially into one type of plastid versus the other. The ability of plastids to import different proteins correlates with the in vivo steady state levels of these proteins. Additional differences were also observed in the intraorganellar portion of the translocation pathway for two thylakoidal proteins. The differences in import characteristics were found to be predominantly governed by information in the transit peptides, since attachment of the various transit peptides to different plastid and foreign proteins demonstrated that the import behavior of the proteins is transferable with the transit sequence. These results indicate that the import mechanisms of leucoplasts and chloroplasts are sufficiently different such that the plastids respond differently to the information present in the transit peptides.

Adenosine Triphosphate↗
PubMed1996

Emerging bacterial zoonotic and vector-borne diseases. Ecological and epidemiological factors.

Among the etiologic agents of emerging infectious diseases are several bacterial organisms that naturally reside in animal and arthropod hosts. The most compelling emerging bacterial zoonotic and vector-borne diseases in the United States are Lyme disease; a Southern erythema migrans-like illness; human monocytic ehrlichiosis; human granulocytic ehrlichiosis; a novel cat flea-associated typhus group rickettsiosis; bartonelloses of immunocompetent and immunocompromised persons, particularly with AIDS; and sylvatic plague. Some of these antimicrobial-treatable infections are life threatening. During the acute stage of illness when antimicrobial agents are most effective, the flulike clinical signs and symptoms and available laboratory tests frequently do not point to a particular diagnosis. Epidemiological factors determined by the ecology of the bacteria are often the most useful diagnostic clues. The recognition of these evolving problems emphasizes the need for development of better laboratory diagnostic methods, for surveillance for and tracking of disease, and for continued research into factors contributing to transmission of the organisms. The continual appearance of previously unidentified bacterial infections requires prospective national strategies for timely recognition of the syndrome, identification of the agent, establishment of criteria and methods for diagnosis, optimization of the treatment regimen, and determination of successful approaches to prevention and control.

Animals↗
PubMed1996

Serodiagnosis of Lyme disease: accuracy of a two-step approach using a flagella-based ELISA and immunoblotting.

An ELISA containing a purified flagellar antigen from Borrelia burgdorferi (FLA-ELISA) was evaluated. The FLA-ELISA, detecting IgM and IgG together, did not have adequate specificity by itself. Good accuracy was obtained, however, when the FLA-ELISA was the first step in a two-step protocol that used immunoblotting as a conditional second test. Samples that scored positive or equivocal by the FLA-ELISA were evaluated with separate IgM and IgG immunoblots. The sensitivity of the two-step process for patients with erythema migrans or with later manifestations of Lyme disease was 64% and 100%, respectively. The specificity for health blood donors was 100% and was 90% for the aggregate of all persons with illness that may cause serologic cross-reactivity (98% if the samples from relapsing fever patients were excluded). Test precision was 96% overall, 99% for Lyme disease case serum samples, 100% for specimens from blood donors, and 88% for samples from persons with other illness.

Antigens, Bacterial↗
PubMed1996

Low seroprevalence of human Lyme disease near a focus of high entomologic risk.

To investigate a low rate of reported human Lyme disease adjacent to an area where the vector tick had become well established, we performed human and canine serosurveys and gathered data on environmental factors related to the risk of transmission. In March 1993, we obtained serum samples and conducted questionnaires that included information on outdoor activities, lot size, and frequency of deer sightings from 272 individuals living within a 5-km strip extending 12 km inland from a study site in south coastal Maine where collections revealed an abundant population of deer ticks. Serologic analysis was done using a flagellin-based enzyme-linked immunosorbent assay (ELISA) followed by Western immunoblot of positive and equivocal samples. Sera from 71 unvaccinated dogs within the study area were also analyzed for anti-Borrelia antibodies by ELISA. Human seropositivity was limited to two individuals living within 1.2 km of the coast. The frequency of daily deer sightings decreased sharply outside this area. Canine seropositivity, 100% within the first 0.8 km, decreased to 2% beyond 1.5 km. Canine serology appears to correlate with the entomologic indicators of the risk of Lyme disease transmission. Possible explanations for the low human seroprevalence are offered.

Adolescent↗
PubMed1995

Import characteristics of a leucoplast pyruvate kinase are influenced by a 19-amino-acid domain within the protein.

Two cDNA clones encoding distinct forms of plastid pyruvate kinase (designated Pka and Pkg) have recently been characterized. Pkg is found in both leucoplasts and chloroplasts, whereas Pka is present only in leucoplasts. The precursors of these proteins have different in vitro import characteristics. The Pkg precursor behaves like a typical stromal protein precusor with both types of plastid. In contrast, Pka precursors accumulate on the outer envelope membrane of leucoplasts under the same assay conditions and require a higher level of ATP for import into the organelle. Interestingly, the binding of Pka precursors to chloroplasts cannot be detected at any tested level of ATP even though the precursors are imported into the organelle at higher concentrations of ATP. Various N-terminal deletions and chimeric fusions were used to examine the translocation signaling mechanism of the Pka precursor. The N-terminal 83-amino-acid segment of Pka contains a transit peptide that is capable of directing dihydrofolate reductase and the mature body of Pkg into both types of plastid. Unlike the complete Pka precursor, these fusion proteins behave like typical stromal protein precursors. The behavior of the Pka transit peptide is influenced by a 19-amino-acid domain (-P-S-S-I-E-V-D-A-V-T-E-T-E-L-K-E-N-G-F-) located immediately downstream of the N-terminal 83-residue segment. Deletion of this domain from Pka alters its import properties such that it resembles a typical stromal protein precursor. Re-introduction of the 19-residue domain into the Dhfr fusion protein alters its import characteristics to resemble that of the complete Pka precursor. This 19-amino-acid domain can also influence the function of transit sequences from other precursors when it is placed immediately behind the transit peptide. These results suggest that this 19-amino-acid domain plays an important role in governing the import characteristics of the Pka precursor. We have named this 19-residue segment the "import modifying domain."

Adenosine Triphosphate↗
PubMed1995

Histidine residues 139, 363 and 500 are essential for catalytic activity of cofactor-independent phosphoglyceromutase from developing endosperm of the castor plant.

Cofactor-independent phosphoglyceromutase (PGM) from castor is inactivated by diethyl pyrocarbonate, implicating histidine residues in the catalytic mechanism. Treatment of the inhibited enzyme with 1 M hydroxylamine at pH 7.0 restores the enzyme activity. Spectroscopic data indicate that the inactivation of PGM with diethyl pyrocarbonate is the result of formation of carbethoxyhistidine derivatives. The substrate, 3-phosphoglycerate, substantially protects the enzyme against diethyl pyrocarbonate inactivation, indicating that the histidine residues important in catalysis are at or near the active site of the enzyme. There are 12 conserved histidine residues in all plant PGMs that have been sequenced. In the castor PGM, these conserved histidine residues were changed to either valine (H12V) or alanine (H41A, H65A, H84A, H127A, H139A, H163A, H363A H433A, H471A, H500A and H540A) by in vitro mutagenesis. Expression of these mutant proteins in Escherichia coli produced seven soluble mutant proteins (mutations H41A, H65A, H84A, H139A, H363A, H500A and H540A) and five insoluble mutant proteins (mutations H12V, H127A, H163A, H433A and H471A). Among the seven soluble proteins, four possessed normal PGM activity (mutations H41A, H65A, H84A and H540A) and three (mutations H139A, H363A and H500A) had no catalytic activity. Along with the in-vitro-expressed wild-type enzyme, mutant enzymes [H139A]PGM, [H363A]PGM and [H500A]PGM were purified to homogeneity. Purified wild-type PGM expressed in E. coli was active and had a Km value very close to that of the enzyme purified from castor endosperm, while the three mutant enzymes remained inactive throughout purification. Therefore, histidine residues 139, 363 and 500 appear to be essential for the catalytic activity of the cofactor-independent enzyme, and may be located at the active site. Hence, although the cofactor-dependent and cofactor-independent PGMs have no homology in their primary amino acid sequences, both enzymes appear to utilize histidine residues to mediate the transfers of proton and phospho groups in the reaction, and thus may be functionally and mechanistically convergent.

Amino Acid Sequence↗
PubMed1995

Structure of the genes encoding the alpha- and beta-subunits of castor pyrophosphate-dependent phosphofructokinase.

Full-length genomic clones encoding the alpha- and beta-subunits of the pyrophosphate-dependent phosphofructokinase (PFP) from the castor plant have been isolated and sequenced. The gene (PFP alpha) encoding PFP alpha is approx. 5.8 kb in length and contains 19 exons, which collectively encode a protein of 617 amino acids (aa) having a deduced M(r) of 67,360. PFP beta is approx. 4.6-kb long and contains 16 exons. Together, these exons encode a protein (PFP beta), of 552 aa with a deduced M(r) of 60,114. The intron-exon splice junctions in both genes contain the consensus sequences typical for plants. An alignment of intron placement in castor PFP alpha and PFP beta with introns in the 5' portion of the gene encoding the ATP-dependent phosphofructokinase (PFK) from rabbit muscle, indicates that only one intron occupies the same position in all three genes. Furthermore, within castor PFP alpha and PFP beta, only two introns are identically placed. Within the promoter regions of castor PFP alpha and PFP beta, there are short sequences having high homology to each other (up to 65%). The results demonstrate, for the first time, that there is little homology between PFP and PFK, nor are PFP alpha and PFP beta closely related. This lack of homology suggests PFP did not evolve from PFK, but rather, that PFP and PFK have probably evolved from a common ancestral gene.

Amino Acid Sequence↗
PubMed1995

Molecular characterization of plastid pyruvate kinase from castor and tobacco.

Clones encoding two different forms of plastid pyruvate kinase (PKp; EC 2.7.1.40) have been isolated from both castor and tobacco seed cDNA libraries. One form, designated PKpA, from castor was described in a previous report, and the tobacco homologue of PKpA has now been isolated. In addition, a second cDNA, designated PKpG, has been identified and sequenced in both species. Western blot analysis, using antibodies raised against protein overexpressed from these clones, indicates that they encode the two predominant polypeptides of plastid pyruvate kinase from developing castor endosperm. In castor, both PKpA and PKpG are encoded by single genes. In the allotetraploid Nicotiana tabacum, there are two copies of each, one derived from each of the progenitors of this species. The expression of the genes for PKpA and PKpG was examined in various tissues from both castor and tobacco. In castor, both forms are expressed in developing and germinating endosperm and in the root but neither is expressed in the leaf. In tobacco, both forms are expressed in developing seeds but in mature tissues, PKpA is most abundant in roots and PKpG in leaves.

Amino Acid Sequence↗
PubMed1995

Biliary complications in the treatment of unsubstantiated Lyme disease.

Treatment of unsubstantiated Lyme disease has led to serious complications in some cases. Two case-control studies, based on information in clinical records of patients discharged with a diagnosis of Lyme disease during 1990-1992, were conducted at a central New Jersey hospital. Twenty-five patients with biliary disease were identified, and 52 controls were selected from 1352 patients with suspected Lyme disease. Only 3% of 71 evaluatable subjects met the study criteria for disseminated Lyme disease. Patients with biliary disease were more likely than were antibiotic controls to have received ceftriaxone and more likely than ceftriaxone controls to have received a daily ceftriaxone dose > or = 40 mg/kg and to be < or = 18 years old. Fourteen of 25 biliary case-patients underwent cholecystectomy; all had histopathologic evidence of cholecystitis and 12 had gallstones. Thus, treatment of unsubstantiated diagnoses of Lyme disease is associated with biliary complications.

Adolescent↗
PubMed1995

Epidemiologic and diagnostic studies of patients with suspected early Lyme disease, Missouri, 1990-1993.

A retrospective case-control study investigated 45 Missouri outpatients with annular rashes meeting a surveillance case definition for erythema migrans and with onset in 1990-1991. Risk factors included being male, living near a body of water, and hunting. Twenty patients (44%) associated their rash with the bite of a tick; of these, 5 described an adult Amblyomma americanum. A typical rash was described as expanding over time and measuring 8 cm in diameter at 4 days after onset. Mild constitutional symptoms were common but fever was uncommon. Serologic tests failed to incriminate Borrelia burgdorferi or selected other arthropodborne pathogens. Skin specimens from suspected erythema migrans lesions of 23 Missouri patients sampled prospectively in 1991-1993 were culture-negative for B. burgdorferi. Thus, tick bite-associated annular rashes in Missouri remain idiopathic. Possible causes include infection with a novel A. americanum-transmitted pathogen and an atypical toxic or immunologic reaction to tick-associated proteins.

Adolescent↗
PubMed1995

Lyme disease.

A clear understanding of the epidemiology of Lyme disease is needed to bring about its effective treatment, prevention, and control. Strategies for effective intervention must be based on quantitative descriptions of the complex interactions of B. burgdorferi with its human and other vertebrate hosts, tick vectors, and the environment. Sound decision making on diagnosis and patient management requires an understanding of the prior probabilities of infection and disease, the performance characteristics of laboratory tests, and the cost-benefits of various treatments. Lyme disease is still a relatively new disease for which there are many unanswered questions; it is an exciting and challenging model of an emerging vector-borne zoonotic disease.

Animals↗