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Biomedical subjects

M D Murphy

Publications and source records attributed to M D Murphy.

At least 19 recordsLinked to original sources

Experimental vesicular stomatitis virus infection in horses: effect of route of inoculation and virus serotype.

Horses were inoculated with Vesicular stomatitis New Jersey and Indiana viruses by routes simulating contact and vector transmission. Clinical signs, lesions, antibody development, viral shedding and persistence, and viremia were monitored. Horses were infected with both viruses by all routes as confirmed by seroconversion. Salivation, primary lesions at inoculation sites, and secondary oral lesions were the most common clinical findings. Viral shedding was most often from the oral cavity, followed by the nasal cavity; titers were highest from oral cavity samples. Virus was rarely isolated from the conjunctival sac and never from feces or blood. Development of neutralizing antibody coincided with cessation of lesion development and detection of virus by isolation. Circulating virus-specific IgM, IgG, IgA, and neutralizing antibodies developed in most animals postinoculation (PI) days 6 to 12, depending on the route of inoculation. At postmortem (PI days 12 to 15), lesions were healing, were not vesicular, and did not contain detectable virus by isolation, reverse transcriptase polymerase chain reaction, or immunohistochemistry. Numerous infiltrating lymphocytes and plasma cells suggested that lesion resolution was partially due to local immunity. Detection of viral RNA from tonsil and lymph nodes of head at necropsy suggests that these tissues play a role in the pathogenesis of the disease; molecular techniques targeting these tissues may be useful for confirming infection in resolving stages of disease. The routes of inoculation used in this study reflect the diversity of transmission routes that may occur during outbreaks and can be used to further study contact and vector transmission, vaccine development, and clarify pathogenesis of the disease in horses.

Animals↗

Failure of porcine reproductive and respiratory syndrome virus to replicate in porcine endothelial cell cultures.

Clinical, gross, and microscopic pathologic and immunohistochemical findings in pigs infected with porcine reproductive and respiratory syndrome virus (PRRSV) suggest that PRRSV may replicate in endothelial cells. Endothelial cell cultures from porcine aorta and pulmonary artery were tested for susceptibility to various strains of PRRSV. Cultures were identified as endothelium by light microscopy and immunohistochemical staining for P-selectin and von Willebrand factor. Five strains of PRRSV, i.e., the National Veterinary Services Laboratories, Ames, IA PRRSV strain 130-PDV and 4 field strains isolated from pneumonic lungs, failed to replicate in these porcine large-vessel endothelial cell cultures.

Animals↗

How shared are age-related influences on cognitive and noncognitive variables?

Several theories have suggested that age-related declines in cognitive processing are due to a pervasive unitary mechanism, such as a decline in processing speed. Structural equation model tests have shown some support for such common factor explanations. These results, however, may not be as conclusive as previously claimed. A further analysis of 4 cross-sectional data sets described in Salthouse, Hambrick, and McGuthry (1998) and Salthouse and Czaja (2000) found that although the best fitting model included a common factor in 3 of the data sets, additional direct age paths were significant, indicating the presence of specific age effects. For the remaining data set, a factor-specific model fit at least as well as the best fitting common factor model. Three simulated data sets with known structure were then tested with a sequence of structural equation models. Common factor models could not always be falsified--even when they were false. In contrast, factor-specific models were more easily falsified when the true model included a unitary common factor. These results suggest that it is premature to conclude that all age-related cognitive declines are due to a single mechanism. Common factor models may be particularly difficult to falsify with current analytic procedures.

Adolescent↗

Are emotionally charged lures immune to false memory?

Using the Deese-Roediger-McDermott task and E. Tulving's (1985) remember-know judgments for recognition memory, the authors explored whether emotional words can show the false memory effect. Participants studied lists containing nonemotional, orthographic associates (e.g., cape, tape, ripe; part, perk, dark) of either emotional (e.g., rape) or nonemotional (e.g., park) critical lures. This setup produced significant false "remembering" of emotional lures, even though initially no emotional words appeared at study. When 3 emotional nonlure words appeared at study, emotional-lure false recognition more than doubled. However, when these 3 study words also appeared on the recognition test, false memory for the emotional lures was reduced. Across experiments, participants misremembered nonemotional lures more often than they did emotional lures, but they were more likely to rate emotional lures as "remembered," once they had been recognized as "old." The authors discuss findings in light of J. J. Freyd and D. H. Gleave's (1996) criticisms of this task.

Adult↗

Contact transmission of vesicular stomatitis virus New Jersey in pigs.

OBJECTIVE: To determine how viral shedding and development or lack of clinical disease relate to contact transmission of vesicular stomatitis virus New Jersey (VSV-NJ) in pigs and determine whether pigs infected by contact could infect other pigs by contact. ANIMALS: 63 pigs. PROCEDURE: Serologically naive pigs were housed in direct contact with pigs that were experimentally inoculated with VSV-NJ via ID inoculation of the apex of the snout, application to a scarified area of the oral mucosa, application to intact oral mucosa, or ID inoculation of the ear. In a second experiment, pigs infected with VSV-NJ by contact were moved and housed with additional naive pigs. Pigs were monitored and sampled daily for clinical disease and virus isolation and were serologically tested before and after infection or contact. RESULTS: Contact transmission developed only when vesicular lesions were evident. Transmission developed rapidly; contact pigs shed virus as early as 1 day after contact. In pens in which contact transmission was detected, 2 of 3 or 3 of 3 contact pigs were infected. CONCLUSIONS AND CLINICAL RELEVANCE: Transmission was lesion-dependent; however, vesicular lesions often were subtle with few or no clinical signs of infection. Contact transmission was efficient, with resulting infections ranging from subclinical (detected only by seroconversion) to clinical (development of vesicular lesions). Long-term maintenance of VSV-NJ via contact transmission alone appears unlikely. Pigs represent an efficient large-animal system for further study of VSV-NJ pathogenesis and transmission.

Animals↗

Misguided multiplication: creating false memories with numbers rather than words.

We built Deese (1959)/Roediger and McDermott (1995) (DRM) false memory lists composed of multiplication problems rather than words. Half these lists contained table-related, near neighbors (e.g., 3 x 7 = ??, 3 x 9 = ??) of a missing multiplication answer lure (e.g., 24). The other half contained problems unrelated to the lure (e.g., 5 x 5 = ??, 11 x 3 = ??). Participants solved each problem in a single list and then took immediate recognition (Experiment 1) or recall and then recognition tests (Experiment 2) for the answers. Many people misremembered that the lure was an answer to a study-phase problem, but only when solving the study list that contained the lure's neighbors. False memory was also greater for some list-lure combinations than others, as seen previously with words. We have thus demonstrated that numbers can also produce false memory, and we use the mental math and DRM task literatures to explain these results.

Adult↗

A beautiful day in the neighborhood: what factors determine the generation effect for simple multiplication problems?

In four experiments, we examined the generation effect for the free recall of simple multiplication answers. Large-product-size problems showed a consistent generation-effect advantage over small-product-size problems, except when each answer was generated twice, via two different sets of operands (Experiment 2). Also, measures of problem-solution time and strategy use accounted for the large-product-size advantage. Across experiments, however, small-product-size problems (but not large-product-size problems) showed considerable variation in the size of their generation effect. We discovered that solving small-product-size problems via direct memory retrieval increased the episodic recall probability of other problems that were near neighbors to the generated answer, and we attribute this result to a spreading activation mechanism in semantic memory. A measure of neighbor activations, combined with RT to solve each problem, accounted for 51% of the observed generation-effect variance.

Adolescent↗

A cytokinesis checkpoint requiring the yeast homologue of an APC-binding protein.

Checkpoint controls ensure that events of the cell-division cycle are completed with fidelity and in the correct order. In budding yeast with a mutation in the motor protein dynein, the mitotic spindle is often misaligned and therefore slow to enter the neck between mother cell and budding daughter cell. When this occurs, cytokinesis (division of the cytoplasm into two) is delayed until the spindle is properly positioned. Here we describe mutations that abolish this delay, indicating the existence of a new checkpoint mechanism. One mutation lies in the gene encoding the yeast homologue of EB1, a human protein that binds the adenomatous polyposis coli (APC) protein, a tumour suppressor. EB1 is located on microtubules of the mitotic spindle and is important in spindle assembly. EB1 may therefore, by associating with microtubules, contribute to the sensor mechanism that activates the checkpoint. Another mutation affects Stt4, a phosphatidylinositol-4-OH kinase. Cold temperature is an environmental stimulus that causes misalignment of the mitotic spindle in yeast and appears to activate this checkpoint mechanism.

1-Phosphatidylinositol 4-Kinase↗

Blastopore formation in the animal hemisphere: functional inversion of gastrulation by centrifugation of Xenopus laevis eggs.

Eggs of Xenopus laevis and many other amphibians contain a gradient of yolk platelets along the animal-vegetal axis. Small platelets predominate in the animal hemisphere, and a boundary between medium and large yolk platelets exists near the equator. The blastopore forms at this boundary at the beginning of gastrulation, in the vegetal hemisphere. Does this boundary have a role in determination of the position of the blastopore, or does cortical information predominate? Past experiments using 1g to invert the egg showed a distinct tendency to form the blastopore in the original vegetal hemisphere. Our experiments, however, have used 20g centrifugation to achieve a more complete inversion of the yolk gradient. The blastopore formed in the original animal hemisphere in >95% of surviving gastrulae, if centrifugation was begun at normalized time 0.20-0.25 of the first cell cycle. Nearly normal larvae (DAI grades 4 and 5) form in 64% of cases (ave. DAI = 3.2). These data support the idea that the position of the blastopore depends on the position of the egg's internal contents rather than cortical determinants.

Animals↗

Determining the requirements for cooperative DNA binding by Swi5p and Pho2p (Grf10p/Bas2p) at the HO promoter.

SW15 encodes a zinc finger DNA binding protein required for the transcription of the Saccharomyces cerevisiae HO gene, and PHO2 encodes a homeodomain DNA binding protein. In vitro biochemical studies using purified Swi5p and Pho2p proteins have demonstrated that Swi5p and Pho2p bind cooperatively to the HO promoter. In this report we investigate the regions of the Swi5p and Pho2p proteins required for cooperative DNA binding. The analysis of each protein gives a similar result: the zinc finger or homeodomain DNA binding domains are each sufficient for in vitro DNA binding, but additional regions of each protein are required for cooperative DNA binding. In vitro and in vivo experiments were conducted with promoters with altered spacing between the Pho2p and Swi5p binding sites. Mutations that increased the distance between the two binding sites had minimal effects on either in vitro cooperative DNA binding or in vivo upstream activating sequence activity. These observations suggest that the interaction domains of Swi5p and Pho2p are flexible and can tolerate an increase in distance between the two binding sites. The mechanism of the cooperative DNA binding by Swi5p and Pho2p is discussed.

Base Sequence↗

Resting energy expenditures measured by indirect calorimetry are higher in preadolescent children with cystic fibrosis than expenditures calculated from prediction equations.

OBJECTIVE: This study compared measured resting energy expenditures to resting energy expenditures calculated using Harris-Benedict equations (HBEs) and the Cystic Fibrosis Consensus Committee equations (CFCCEs). DESIGN: We studied 31 preadolescent boys and girls with cystic fibrosis who ranged in age from 3.25 to 12.75 years old. The patients were afebrile and not in pulmonary distress. Measured resting energy expenditures were determined using a portable metabolic measurement cart with fully automated calibration and data management. The measured resting energy expenditures obtained were compared with values obtained using HBEs and CFCCEs. RESULTS: For each patient, the measured resting energy expenditure value was above the predicted resting energy expenditure values derived from HBEs (P < or = .0001) and CFCCEs (P < or = .01). APPLICATIONS: The HBEs and the CFCCEs underestimated the energy expenditures of the study population by 13% and 8%, respectively. These findings support the usefulness of the measurement of energy expenditures in determining the energy needs of preadolescent patients with cystic fibrosis. In clinical practice, the resting energy expenditures would be multiplied by activity coefficients to determine the total daily energy expenditures of this population.

Basal Metabolism↗

S-laminin expression in adult and developing retinae: a potential cue for photoreceptor morphogenesis.

The development of the neural retina follows a stereotyped time course that begins with an undifferentiated neuroepithelium populated by multipotential progenitor cells and ends with a highly differentiated tissue containing diverse cell types. The identities of the factors that guide this differentiation have remained elusive; a likely location for such factors, however, is the extracellular environment. Here, we show that the extracellular matrix component s-laminin is present in the neural retina, that s-laminin expression parallels the differentiation of rod photoreceptors, that photoreceptors interact with s-laminin in vitro, and that antibodies to s-laminin profoundly reduce the appearance of cells that express rhodopsin in vitro. These data suggest that s-laminin plays a role in the differentiation of the neural retina and provide evidence that the composition of the extracellular matrix may be an important determinant of retinal differentiation.

Animals↗

Adult age comparisons in the processing of event frequency information.

The usual superiority in frequency-of-occurrence judgments of younger vs. older subjects was hypothesized to result from greater strategic encoding of the materials conveying frequency information. A subject-paced, visual search task was designed to control nontarget word encoding. Relative frequency judgments for the nontarget word pairs were equally accurate for younger and older subjects, and performance of both groups was above chance. Results suggest that strategic cover-task encoding can induce age differences in incidental frequency processing. Consistent with a nonoptimal as opposed to an optimal view, automatic processes require only minimal capacity for above-chance performance, but additional strategic resources can increase performance. When such additional resources are used more by younger than by older subjects, the former are better in frequency performance.

Adult↗

Pseudomonas aeruginosa flagellar antibodies in patients with cystic fibrosis.

An enzyme-linked immunosorbent assay specific for flagellum type (a or b) of Pseudomonas aeruginosa was used to detect serum immunoglobulin antibodies in 98 random outpatients and 14 colonized cystic fibrosis patients. Antibodies were detected to both types of flagella in addition to M-2 lipopolysaccharide. Titers to both flagellar antigens (FlAg) were 10 to 100 times higher in cystic fibrosis patients than in random outpatients of a comparable age group. Mean antibody titers against b-type FlAg were 454 for outpatients (ages newborn to 21 years), whereas the mean titer for cystic fibrosis patients (ages 6 to 21 years) was 51,520. Titers against a-type FlAg were generally lower, with mean outpatient titers of 68 and mean cystic fibrosis patient titers of 34,323. Differences were also seen in antibody titer against M-2 lipopolysaccharide, but these differences did not correspond to M-2 FlAg titers. In 98 random outpatients (ages newborn to 86 years), FlAg titers generally increased with age. To demonstrate further specificity of the enzyme-linked immunosorbent assay for flagellum antibody, Western blots were performed with selected high-titer cystic fibrosis patient sera. Sera that had a high titer (greater than 25,600) for b- or a-type FlAg showed a corresponding reactive band. These results demonstrate that flagellum antibodies are produced in humans in response to P. aeruginosa infection.

Adolescent↗

Evaluation of a monoclonal antibody pool for rapid diagnosis of respiratory viral infections.

A pool of monoclonal antibodies (MAbP) was evaluated both as a method of cell culture confirmation and as a rapid diagnostic screen for viral infection in respiratory secretions. The MAbP was used in a two-step fluorescent staining procedure on cells harvested from cultures (phase 1) and on exfoliated nasopharyngeal or tracheal cells (phase 2). Antibodies in the MAbP were directed against respiratory syncytial virus, adenoviruses, parainfluenza virus types 1, 2, and 3, and influenza viruses A and B. Individual antiviral antibody stains were used to identify specific viruses from MAbP-positive specimens. In phase 1 (cell culture confirmation only), 241 respiratory specimens were tested. MAbP sensitivity and specificity were 91 and 94%, respectively. In phase 2, 376 respiratory specimens were evaluated by direct staining of exfoliated cells, and these results were compared with results of cell culture isolation. The sensitivity and specificity of the MAbP used in direct specimen testing were 69 and 97%, respectively. These results indicate that the MAbP is highly specific and fairly sensitive for detection of seven different respiratory viruses in one testing procedure. The MAbP is a rapid screening technique for respiratory secretions and is potentially a cost-effective approach to viral detection.

Adenoviruses, Human↗