Reasons to doubt the present evidence for metaphoric representation.
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Cognition↗
Biomedical subjects
G L Murphy
Publications and source records attributed to G L Murphy.
Characterization of a new isolate of Ehrlichia platys (Order Rickettsiales) using electron microscopy and polymerase chain reaction.
A mixed-breed pup approximately 3 months old obtained in north central Oklahoma by the Laboratory Animal Resources Unit of Oklahoma State University presented with platelet inclusions. The dog developed severe thrombocytopenia (< 10,000 microliters-1) following the appearance of inclusions. Blood films were monitored daily and when about 75% of platelets had inclusions, samples were collected in EDTA and processed for electron microscopic (EM) studies and polymerase chain reaction (PCR). EM studies on glutaraldehyde-fixed buffy coat revealed rickettsia-like inclusions in numerous platelets. Serologic examination, using Ehrlichia platys antigen, showed high titre suggestive of E. platys infection. PCR primers derived from a highly variable region of the 16S rRNA gene sequence of E. platys were used to specifically amplify that region of the parasite's DNA. Sequencing of the PCR product obtained by general Ehrlichia primers showed one nucleotide difference from the published sequence for E. platys which suggests possible strain variation of this intracellular parasite. Our results indicate that PCR may be a useful tool in the diagnosis of E. platys infection and that, like other Ehrlichia spp., E. platys isolates may vary.
Animals↗
Outer membrane proteins of bovine Pasteurella multocida serogroup A isolates.
The outer membrane proteins (OMPs) of P. multocida serotypes A3 (7 isolates), A4 (2 isolates), A3,4 and A2 (one isolate each) obtained from pneumonic cattle (10 isolates) and from one pig isolate were investigated to identify potential immunogens. SDS-PAGE of P. multocida OM isolated by SDG centrifugation of spheroplasts revealed eight major OMPs. Outer membranes isolated by sarcosyl extraction or SDG had similar protein composition on Coomassie blue-stained SDS-PA gel and on immunoblots. Two major OMPs (M(r)s of 35 and 46 kDa at 100 degrees C) demonstrated heat modifiability with apparent M(r)s of 30 and 34 kDa at 37 degrees C, respectively. The N-terminal aa sequences of these heat modifiable proteins revealed homology with E. coli OmpA and Hib P1 proteins, respectively. Protease treatment of whole cells followed by western immunoblots using bovine convalescent sera identified several immunogenic, surface-exposed and conserved OMPs among the eleven P. multocida isolates examined. The whole organism SDS-PAGE profiles of the eleven P. multocida isolates differed such that six patterns were seen. These patterns could potentially be used as a typing system for P. multocida bovine isolates based on the molecular weights of whole cell proteins. The above observations have potentially important implications relative to the immunity to infection.
Amino Acid Sequence↗
The effects of prior processing episodes on basic-level superiority.
Four experiments were conducted to investigate the effects of prior processing episodes on people's preference for categorizing objects at the basic level (e.g. dog) relative to their preference for categorizing at the superordinate (e.g. animal) and the subordinate (e.g. Dalmation) levels. The prior processing episode in Experiment 1 was designed to induce subjects to activate representations at the superordinate level, and those in the remaining experiments were designed to induce subjects to differentiate objects at the subordinate level. After the prior processing episodes, subjects performed either a free naming or a picture categorization task that required them to decide whether an illustrated object belonged to a specified category. Results showed that prior processing episodes modestly reduced the superiority of basic level to superordinate level and subordinate level in categorization but not in free naming. The results suggest that the basic-level advantage is subject to the effects of context, but the effects are not as strong as the context effects on other aspects of categorization behaviour (e.g. rating typicality of a category member). Hence, the preference for the basic level is a somewhat more stable, invariant aspect of conceptual representation. Possible determinations of this stability are discussed.
Cognition↗
Purification and partial characterization of the OmpA family of proteins of Pasteurella haemolytica.
This study was conducted to partially characterize and identify the purity of two major outer membrane proteins (OMPs) (with molecular weights of 32,000 and 35,000 [32K and 35K, respectively]) of Pasteurella haemolytica. The 35K and 32K major OMPs, designated Pasteurella outer membrane proteins A and B (PomA and PomB, respectively), were extracted from P. haemolytica by solubilization in N-octyl polyoxyl ethylene. The P. haemolytica strain used was a mutant serotype A1 from which the genes expressing the 30-kDa lipoproteins had been deleted. PomA and PomB were separated and partially purified by anion-exchange chromatography. PomA but not PomB was heat modifiable. The N-terminal amino acid sequences of the two proteins were determined and compared with reported sequences of other known proteins. PomA had significant N-terminal sequence homology with the OmpA protein of Escherichia coli and related proteins from other gram-negative bacteria. Moreover, polyclonal antiserum raised against the E. coli OmpA protein reacted with this protein. PomA was surface exposed, was conserved among P. haemolytica biotype A serotypes, and had porin activity in planar bilayers. No homology between the N-terminal amino acid sequence of PomB and those of other known bacterial proteins was found. Cattle vaccinated with live P. haemolytica developed a significant increase in serum antibodies to partially purified PomA, as shown by enzyme-linked immunosorbent assays, and to purified PomA and PomB, as detected on Western blots and by densitometry.
Amino Acid Sequence↗
Use of a nonradioactive DNA probe for detection of Anaplasma marginale infection in field cattle: comparison with complement fixation serology and microscopic examination.
A sensitive Anaplasma marginale-specific 409-base pair DNA probe was developed in a previous study for detection of A. marginale infection in experimentally infected cattle with a test that employed slot-blot and in situ hybridization. To test the suitability of the probe to detect A. marginale in the blood of naturally infected carrier cattle, slot-blot hybridization was used to determine the infection rate of A. marginale in cattle from 3 geographic areas in Oklahoma. For comparison, blood samples from the same cattle were also examined by light microscopy and were tested by the complement fixation test. For the DNA hybridization assay, the probe was labeled with digoxigenin 11-dUTP by polymerase chain reaction (PCR). DNA was extracted from blood using the QIAamp blood kit and then applied to a nylon membrane and hybridized with the probe. The study herds consisted of 31 beef cows in Harper County, OK, and 42 and 70 dairy cows from Payne and Pittsburg counties, OK, respectively. In the 3 herds, 80.6%, 92.8%, and 57.1% of the cows were positive for A. marginale as assessed with the DNA hybridization assay. In contrast, only 25.8% and 2.86% were complement fixation positive in 2 herds, and no complement fixation positives were found in 1 herd. Uncountable parasitemia that was too low to accurately determine (< 0.01%) from 29.0%, 4.8%, and 11.4% of the samples, respectively, was demonstrated by microscopic examination. All samples positive by complement fixation and microscopic examination had positive probe reactions in the DNA hybridization assay. Therefore, the PCR-mediated nonradioactive DNA probe described here may be useful in epidemiologic investigations and in identification of carrier cattle. This assay could be adapted for use in diagnostic laboratories because it is sensitive, specific, nontoxic, quickly executed, and inexpensive.
Anaplasma↗
Laughter in the cockpit: gelastic seizures--a case report.
We present a case of gelastic seizures in a student naval aviator. He was noted to have uncontrollable fits of laughter on several occasions, but was not referred to his flight surgeon until he had a gelastic seizure while flying in formation, which jeopardized the safety of the flight. He had an aura consisting of lack of concentration, which was then followed by 10 s or less of hysterical laughter. For the previous year and a half, he had had frequent episodes of nocturnal laughter so loud that he woke members of his household and occasionally himself. His neurological evaluation was normal, except for an electroencephalogram (EEG) and a separate video recording, which documented the ictal nature of his events. Gelastic seizures have not previously been discussed in the literature of aerospace medicine. This case illustrates a rare condition that should be considered in patients presenting with inappropriate laughter, and serves as a reminder of the need for continuous, ongoing evaluation of all aircrew by the cognizant flight surgeon.
Adult↗
Differential expression of tubulin isotypes during the cell cycle.
Microtubules play an essential role in cell division. Little is known about possible variations of total tubulin and tubulin isotype expression during the cell cycle. We analyzed the total tubulin content, tubulin polymerization status and tubulin isotype content in resting and dividing human K562 leukemic cells and human MES-SA sarcoma cells. Although the total cellular tubulin content increases as the cells progress toward mitosis, the total tubulin/total protein ratio is stable during the cell cycle. Reverse transcriptase-polymerase chain reaction was applied to analyze the levels of expression of alpha, beta, and gamma-tubulin isotypes. Whereas alpha-tubulin isotype and gamma-tubulin transcripts were found to be expressed at constant levels throughout the cell cycle, some of the beta-tubulin isotype transcripts were found to be more highly expressed in dividing then in resting cells. Both of the class IV beta-tubulin isotype transcripts (human 5 beta and beta 2, Class IVa and IVb, respectively) were expressed in dividing K562 and MES-SA cells at twice the levels found in resting cells. Increased expression of the class IV isotype proteins in dividing cells was confirmed by immunoblotting, both in K562 and in MES-SA cells. A larger fraction of total cell tubulin was found to be polymerized in dividing cells (36-40%) than in resting cells (27-30%). The degree of polymerization of class IV tubulin in dividing and resting cells was similar to that of total tubulin. These results show that total tubulin is expressed as constant levels throughout the cell cycle but that the degree of polymerization is increased as cells are committed to division. The relative overexpression of the two class IV beta-tubulin isotypes in dividing cells suggests functional specificity for these isotypes and a regulatory role of these isotypes on the microtubule network during mitosis.
Cell Cycle↗
On metaphoric representation.
The article discusses claims that conceptual structure is in some part metaphorical, as identified by verbal metaphors like LOVE IS A JOURNEY. Two main interpretations of this view are discussed. In the first, a target domain is not explicitly represented but is instead understood through reference to a different domain. For example, rather than a detailed concept of love per se, one could make reference to the concept of a journey. In the second interpretation, there is a separate representation of love, but the content of that representation is influenced by the metaphor such that the love concept takes on the same structure as the journey concept. It is argued that the first interpretation is not fully coherent. The second interpretation is a possible theory of mental representation, but the article raises a number of empirical and theoretical problems for it. It is concluded that many of the data cited as evidence for metaphoric representations can be accounted for by structural similarity between domains.
Concept Formation↗
Genetic and immunological analyses of a 38 kDa surface-exposed lipoprotein of Pasteurella haemolytica A1.
Pasteurella haemolytica serotype A1 is the bacterial pathogen most frequently isolated from the lungs of cattle with bovine respiratory disease. As part of a study to characterize P. haemolytica antigens which are important in eliciting resistance to pneumonic pasteurellosis, we have cloned and sequenced the gene encoding a 38 kDa lipoprotein, Lpp38. The deduced amino acid sequence of Lpp38 is similar to those of the Escherichia coli polyamine transport proteins PotD (70%) and PotF (33%). P. haemolytica Lpp38 is present in both inner membrane and outer membrane fractions of the cell envelope. Susceptibility of Lpp38 to cleavage by extracellular proteases indicates that portions of the protein are surface-exposed. A protein of similar molecular mass in P. haemolytica strains from all 12 serotypes of biotype A and in an untypeable strain was detected by an anti-Lpp38 monoclonal antibody. Lpp38 is recognized by sera from calves resistant to infection after natural exposure to P. haemolytica and by sera from calves protected against infection by vaccination with P. haemolytica A1 outer membranes or with live bacteria. These data suggest a role for this protein in the development of immunity to P. haemolytica infection.
Amino Acid Sequence↗
Developmental studies of Anaplasma marginale (Rickettsiales:Anaplasmataceae) in male Dermacentor andersoni (Acari:Ixodidae) infected as adults by using nonradioactive in situ hybridization and microscopy.
The development of Anaplasma marginale Theiler was studied in ticks using a nonradioactive in situ hybridization method developed in our laboratory. Male Rocky Mountain wood ticks, Dermacentor andersoni Stiles, were infected intrastadially by allowing them to feed for 7 d on an infected calf (acquisition feeding). The ticks were then removed and held in a humidity chamber for 5 d before being fed on a 2nd susceptible call for 10 d (transmission feeding). Two groups of 10 ticks were collected daily during the 22-d experiment. In one group one-half of each tick was processed and embedded in paraffin and in the other group one-half of each tick was embedded in LR White for in situ hybridization. The companion tick halves from each group were fixed and embedded in Dow Epoxy Resin resin for routine light and electron microscopy. As detected by in situ hybridization on LR White- and paraffin-embedded sections and by microscopy, initial infection of A. marginale in ticks occurred in gut tissues either on the 7th d of acquisition feeding or the 1st d of the held period and infection persisted throughout transmission feeding. The highest number of ticks with gut infection was observed on the 5th d of transmission feeding. Salivary glands became infected with A. marginale on the 1st day of transmission feeding and remained infected throughout the transmission feeding period. Peak infection was observed on day 4 of transmission feeding. After the beginning of transmission feeding, A. marginale infection was also observed in interstitial, reproductive, skeletal muscle, fat body, and Malpighian tubule tissues. Although A. marginale infection of ticks clearly originates in midgut epithelial cells, many tissues eventually become infected during transmission feeding, resulting in a generalized infection. The infection of multiple tissues may contribute to the ability of A. marginale infection to persist in intrastadially infected male ticks.
Anaplasma↗
Category-based predictions: influence of uncertainty and feature associations.
Four experiments examined how people make inductive inferences using categories. Subjects read stories in which 2 categories were mentioned as possible identities of an object. The less likely category was varied to determine if people were using it, as well as the most likely category, in making predictions about the object. Experiment 1 showed that even when categorization uncertainty was emphasized, subjects used only 1 category as the basis for their prediction. Experiments 2-4 examined whether people would use multiple categories for making predictions when the feature to be predicted was associated to the less likely category. Multiple categories were used in this case, but only in limited circumstances; furthermore, using multiple categories in 1 prediction did not cause subjects to use them for subsequent predictions. The results increase the understanding of how categories are used in inductive inference.
Adult↗
Antibody responses of cattle to outer membrane proteins of Pasteurella multocida A:3.
OBJECTIVE: To quantify the serum antibody responses to Pasteurella multocida A:3 outer membrane proteins (OMP) for cattle vaccinated with the homologous serogroup and to correlate those responses with the extent of experimentally induced pneumonia. ANIMALS: 29, 5- to 8-month-old beef-type calves. PROCEDURE: Calves were vaccinated SC or by aerosal exposure on days 0 and 7 with live or killed P multocida or phosphate-buffered saline solution (control) and subsequently challenge exposed with virulent P multocida. Antibody responses to P multocida A:3 outer membranes were quantified, using an ELISA. Antibody responses to individual OMP were detected by immunoblot analysis (western blot) and were quantified by densitometry. Antibody responses were compared among groups of calves and for various times after vaccination. Regression analyses were used to determine whether significant correlations existed between lesion scores and antibody responses to either whole outer membranes or to individual OMP. RESULTS: By ELISA, antibody responses to outer membranes for calves aerosol vaccinated with live P multocida were significantly (P < 0.05) greater than those for control calves or for killed P multocida vaccinates. There was a significant (P < 0.05) correlation between lesion score and antibody responses to outer membranes. By western blotting and densitometry, antibodies to 11 prominent OMP (100, 97, 90, 85, 74, 53, 46, 35, 32, 21, and 16 kd) were identified and quantified. In experiment 1, SC vaccination with live P multocida increased antibody binding to all protein bands except 85-, 74-, and 35-kd bands. Aerosol vaccination with live P multocida stimulated increases in antibody binding to all bands except 100 and 16 kd. Antibody responses to the 97-, 90-, 74-, and 35- kd bands were significantly (P < 0.05; greater for live aerosol vaccinates than for control calves. In experiment 2, antibody responses were not different between the killed P multocida vaccinates or control calves Antibody responses for live P multocida aerosol vaccinates were significantly (P < 0.05) greater than those for control calves for the 100-, 90-, 85-, 74-, 53-, 35-, and 16-kd bands. Regression analyses indicated significant correlations (P < 0.05) between lesion score and antibody responses to the 100-, 90-, 53-, 46-, 35-, and 32-kd OMP. CONCLUSIONS: Several OMP of P multocida type A:3 may be important for stimulating immunity to the organism in cattle.
Aerosols↗
Attempted transmission of Ehrlichia canis by Rhipicephalus sanguineus after passage in cell culture.
OBJECTIVES: To compare the transmissibility by the brown dog tick, Rhipicephalus sanguineus, of a recent isolate of Ehrlichia canis (Ebony) with that of another isolate (Oklahoma) that had been passaged in cell culture, and to assess the genetic similarity of the 2 isolates as reflected in the nucleotide (NT) sequence of 16S rDNA. ANIMALS: 13 healthy dogs of various ages and breeds. PROCEDURE: Larval and nymphal ticks were acquisition fed on acutely infected dogs, and, after molting, they were transmission fed as nymphs and adults, respectively, on Ehrlichia-naive dogs. All dogs were monitored daily by blood smear evaluation for evidence of parasitized leukocytes and by physical examination for clinical signs of ehrlichiosis. Serologic and hematologic values were measured weekly. Using a nested polymerase chain reaction, the 16S rDNA was amplified, and the NT sequence of the template DNA was determined. RESULTS: The Ebony isolate of E canis was successfully transmitted to dogs by nymphal and adult ticks. In contrast, no ticks that fed on dogs harboring the cell-cultured isolate (Oklahoma) transmitted it to dogs. On the basis of 16S rDNA sequence, the 2 isolates were 99.9% similar, with only 1 NT difference. CONCLUSIONS: These results reconfirm the vector potential of R sanguineus for E canis. Passage of the Oklahoma isolate of E canis in cell culture apparently adversely affected its transmissibility by ticks, raising the possibility that cell-cultured isolates of this rickettsia may lose their affinity for ticks. Determination of 16S rDNA sequence suggests minor strain variation within the species E canis.
Animals↗
Predicting features for members of natural categories when categorization is uncertain.
An important function of concepts to allow prediction of unseen features. A Bayesian account of feature prediction suggests that people will consider all the categories an object could belong to when they judge the likelihood that the object has a feature. The judgment and decision literature suggests that they may instead use a simpler heuristic in which they consider only the most likely category. In 3 experiments, no evidence was found that participants took into account alternative categories as well as the most likely one when they judged feature probabilities for familiar objects in meaningful contexts. These results, in conjunction with those of Murphy and Ross (1994), suggest that although people may consider alternative categories in certain limited situations, they often do not. Reasons for why the use of alternative categories may be relatively rare are discussed, and conditions under which people may take alternative categories into account are outlined.
Bayes Theorem↗
Hemolytic activity of the Pasteurella haemolytica leukotoxin.
A Pasteurella haemolytica mutant incapable of producing leukotoxin was created by allelic replacement. Concentrated culture supernatants from wild-type P. haemolytica, but not from the mutant, contained the 102-kDa leukotoxin protein and lysed bovine lymphoma cells and sheep erythrocytes. Wild-type P. haemolytica demonstrated the typical beta-hemolytic phenotype on sheep and rabbit blood agar, whereas the mutant did not.
Animals↗
Detection of Anaplasma marginale DNA in bovine erythrocytes by slot-blot and in situ hybridization with a PCR-mediated digoxigenin-labeled DNA probe.
A 409-base pair (bp) DNA fragment derived from the msp-1 beta gene of Anaplasma marginale was amplified and simultaneously labeled with digoxigenin-11-dUTP by a polymerase chain reaction (PCR) assay. The resulting digoxigenin-labeled 409-bp PCR product was used as a probe for slot-blot and in situ hybridization to detect A. marginale DNA from experimentally infected bovine erythrocytes. The hybrid formation was detected with alkaline phosphatase-conjugated anti-digoxigenin antibody and substrates 5-bromo-4-chloro-3-indolyl phosphate and nitroblue tetrazolium salt. In slot-blot hybridizations, the probe detected A. marginale DNA from approximately 1,000-10,000 infected erythrocytes in 1.25 ml of whole blood, which is equivalent to a parasitemia level of 0.00001%. The probe proved to be a A. marginale-specific when tested with 17 species of microorganisms. The applicability of the probe for diagnosis was tested by screening A. marginale infections in 2 experimentally infected splenectomized cattle before microscopically detectable parasitemias and after acute infection. After inoculation of infected blood, A. marginale infections were detected with the probe 14 days prior to detection in stained smears. Microscopically inapparent parasitemias were also detected with the probe for 2 months after acute disease. When the probe was used for in situ hybridization on methanol-fixed blood smears, probe reaction could be visualized with light microscopy on A. marginale inclusions within infected erythrocytes. The probe reaction was not observed on leukocytes and uninfected erythrocytes from infected blood smears, on erythrocytes from uninfected blood samples, or on samples infected with A. ovis, Babesia bovis, or B. bigemina. This PCR-mediated nonradioactive probe appears to be a sensitive diagnostic test for A. marginale.
Anaplasma↗
Construction of isogenic mutants of Pasteurella haemolytica by allelic replacement.
We describe methods for the mutagenesis of cloned Pasteurella haemolytica (Gram-) genes and for the construction of P. haemolytica mutants by allelic exchange. We used these methods to construct isogenic mutants of P. haemolytica which no longer synthesize three membrane lipoproteins (Lpp). A single genetic locus, consisting of three tandemly arranged genes encoding 28-30-kDa membrane Lpp, was replaced with a mutated locus which carries the beta-lactamase-encoding ApR gene from a 4.2-kb P. haemolytica plasmid. The inactivated locus was introduced into P. haemolytica by electroporation of a plasmid which carries the mutated locus, but is incapable of replicating in P. haemolytica. Southern and Western blot analyses indicate that the wild-type locus was replaced by the mutated locus through a double-crossover recombination event and that the membrane Lpp were no longer produced by the mutant strain. These methods should be useful in constructing mutant loci which can be used to analyze the roles for various P. haemolytica proteins in the pathogenesis of bovine pneumonic pasteurellosis.
Alleles↗