PCR detection of North American and Central African isolates of epizootic hemorrhagic disease virus (EHDV) based on genome segment 10 of EHDV serotype 1

PCR amplification technology for the detection of epizootic hemorrhagic disease virus (EHDV) ribonucleic acid in cell culture and clinical specimens was developed. With oligoribonucleotide primers selected from genome segment 10 of EHDV serotype 1 (EHDV-1), which codes for two nonstructural proteins (NS3 and NS3a), the PCR-based assay resulted in a 535-bp PCR product. RNAs from North American EHDV-1 prototype, EHDV-2 prototype, and a number of EHDV field isolates, including the Central African isolates of EHDV-5 and EHDV-318 propagated in cell cultures, were detected by this PCR-based assay. The specific 535-bp PCR products were visualized onto agarose gels, and the identity of the PCR products was confirmed by chemiluminescent hybridization with a 352-bp internal probe. The sensitivity of the EHDV PCR assay was increased by chemiluminescent hybridization; by this EHDV-NS3 PCR, 10 fg of EHDV RNA was detected (equivalent to 600 viral particles). Amplification product was not detected when the PCR-based assay was applied to RNAs from North American bluetongue virus prototype serotypes 2, 10, 11, 13, and 17; total nucleic acid extracts from uninfected BHK-21 cells; or unfractionated blood from calves and deer that were EHDV seronegative and virus isolation negative. The described EHDV PCR-based assay with primers derived from segment 10 of EHDV-1 resulted in detection of EHDV RNA from blood and tissues collected from calves and deer with natural and experimental EHDV infections and provides a valuable tool to study the epidemiology of EHDV infection in susceptible ruminants.     

Isolation of cell cycle-dependent gamma ray-sensitive Chinese hamster ovary cell

A technique for the isolation of gamma ray-sensitive Chinese hamster ovary (CHO) cell mutants is described, which uses nylon cloth replica plating and photography with dark-field illumination to directly monitor colonies for growth after gamma irradiation. Two gamma ray-sensitive mutants were isolated using this method. One of these cells (XR-1) had a two-slope survival curve: an initial steep slope and then a flattening of the curve at about 10% survival. Subsequently, it was found that this cell is sensitive to gamma irradiation in G1, early S, and late G2 phases of the cell cycle, whereas in the resistant phase (late S phase) its survival approaches that of the parental cells. The D37 in the sensitive G1 period is approximately 30 rads, compared with 300 rads of the parental cell. This mutant cell is also sensitive to killing by the DNA breaking agent, bleomycin, but is relatively insensitive to UV light and ethyl methane sulfonate, suggesting that the defect is specific for agents that produce DNA strand breakage.     

Arrest of B lymphocyte terminal differentiation by CD40 signaling: mechanism for lack of antibody-secreting cells in germinal centers

Dendrimers represent a new class of highly branched polymers whose interior cavities and multiple peripheral groups facilitate potential applications in biomedicine and bio-organic chemistry. Major advances in the past year were made in the synthesis and study of new carbohydrate, nucleic acid, and peptide dendrimers, as well as in the use of dendrimers as magnetic resonance imaging contrast agents, as agents for cellular delivery of nucleic acids, and as scaffolds for biomimetic systems.     

A 1957 outbreak of Legionnaires' disease associated with a meat packing plant

Retrospective study shows that a 1957 outbreak of pneumonia in Austin, Minnesota, was Legionnaires' disease. Between June 7 and August 9, 1957, 78 persons were hospitalized with acute respiratory disease of unknown cause. Most had fever, headache, cough, and pneumonitis; two died. Ages ranged from 14-83 years; half of the patients were aged 55 years or older. Eighty-seven per cent were men. There were no secondary cases. Forty-six (59%) of the 78 patients were employees at a local meat packing plant, in distinction to the area's total working population (32%). Serosurvey of 15 of the 1957 outbreak cases and 30 controls matched for age, sex, and either occupation or residence was carried out in 1979. Antibody titers were determined for Legionella pneumophila serogroups 1-4 by means of indirect immunofluorescence. Twelve (80%) of the 15 cases and 13 (43%) of the 30 controls had antibody titers of 1:64 or greater to one or more of the L. pneumophila serogroups. Significant differences in L. pneumophila antibody titers (prevalence and level) were found between cases and control groups matched for residence (serogroups 1-3) or occupation (serogroups 2 and 3). Only three of 20 Austin residents with pneumonia diagnosed between 1978 and 1980 had L. pneumophila antibody titers of 1:128 or greater (p less than 0.001), in comparison to cases. These serologic data and the 1957 clinical and epidemiologic observations support the contention that this is the earliest documented outbreak of Legionnaires' disease.     

Randomized trial of an oral platelet glycoprotein IIb/IIIa antagonist, sibrafiban, in patients after an acute coronary syndrome: results of the TIMI 12 trial. Thrombolysis in Myocardial Infarction

In fMRI studies, Gaussian filtering is usually applied to improve the detection of activated areas. Such lowpass filtering enhances the signal to noise ratio. However, undesirable secondary effects are a bias on the signal shape and a blurring in the spatial domain. Neighboring activated areas may be merged and the high resolution of the fMRI data compromised. In the temporal domain, activation and deactivation slopes are also blurred. We propose an alternative to Gaussian filtering by restoring the signal using a spatiotemporal Markov Random Field which preserves the shape of the transitions. We define some interaction between neighboring voxels which allows us to reduce the noise while preserving the signal characteristics. An energy function is defined as the sum of the interaction potentials and is minimized using a simulated annealing algorithm. The shape of the hemodynamic response is preserved leading to a better characterization of its properties. We demonstrate the use of this approach by applying it to simulated data and to data obtained from a typical fMRI study.     

Phenoxybenzamine-a mediator of blood oxygen affinity

Studies of the effect of phenoxybenzamine on oxygen transport have been concentrated on flow and resistance changes. Previous studies have ignored the possible effects of phenoxybenzamine on the ability of red blood cells to offload oxygen itself. After an initial inhibitory effect phenoxybenzamine greatly enhances the ability of the red cell to offload oxygen under conditions seen at the tissue level. The disparity between P50std and organic phosphate changes found in this and other studies warrants extreme caution to be used in interpreting results where P50 itself is not measured.     

Secundum atrial septal defect. Nonoperative closure during cardiac catheterization

A 17-year-old girl had clinical and cardiac catheterization findings compatible with a secundum atrial septal defect. During cardiac catheterization, the atrial septal defect was sized and closed using a transvenous umbrella technique.