Clients' interpretation of risks provided in genetic counseling.

Clients in 544 genetic counseling sessions who were given numeric risks of having a child with a birth defect between 0% and 50% were asked to interpret these numeric risks on a five-point scale, ranging from very low to very high. Whereas clients' modal interpretation varied directly with numeric risks between 0% and 15%, the modal category of client risk interpretation remained "moderate" at risks between 15% and 50%. Uncertainty about normalcy of the next child increased as numeric risk increased, and few clients were willing to indicate that the child would probably or definitely be affected regardless of the numeric risk. Characteristics associated with clients' "pessimistic" interpretations of risk, identified by stepwise linear regression, included increased numeric risk, discussion in depth during the counseling session of whether they would have a child, have a living affected child, discussion of the effects of an affected child on relationships with client's other children, and seriousness of the disorder in question (causes intellectual impairment). Client interpretations are discussed in terms of recent developments in cognitive theory, including heuristics that influence judgments about risks, and implications for genetic counseling

Histological Observation of Islet Hemorrhage Induced by Diagnostic Ultrasound with Contrast Agent in Rat Pancreas

Contrast enhanced diagnostic ultrasound CEDUS has been shown to induce capillary hemorrhage in heart and kidney. This study characterized the capillary hemorrhage induced in rat pancreas. The pancreata of anesthetized hairless rats were accessed by laparotomy. A 1.5 MHz diagnostic ultrasound probe with 2.3 MPa peak rarefactional pressure amplitude and 1 s intermittent trigger was used to scan the pancreas, located at the focus (3.8 cm), through saline coupling. The probe was swept to expose the entire organ in 5 min during infusion of Definity® contrast agent at 10 µL/kg/min, and this was repeated in a reverse sweep. The entire pancreas was removed, spread flat for fixation and histological slides were prepared from the mid-plane. Slides were scored blind for islet hemorrhage over the entire area of the organ. Intra-islet microlesions were evident and hemorrhage surrounded many islets. The hemorrhage often impacted nearby acini, and expanded into inter-lobular septa. In CEDUS pancreata removed soon after scanning, 76.2±11.8% (n = 6) of islets had evidence of hemorrhage and/or islet microlesions compared to 1.1±2.5% (n = 5) for sham CEDUS (P<0.001). In pancreata removed after 4 hr, fibrin formation was detected by immunohistology in the hemorrhage and intra-islet microlesions. Diagnostic ultrasound with contrast agent induced substantial capillary hemorrhage in rat pancreas, concentrated particularly in the islets

Efficient Cellular Release of Rift Valley Fever Virus Requires Genomic RNA

The Rift Valley fever virus is responsible for periodic, explosive epizootics throughout sub-Saharan Africa. The development of therapeutics targeting this virus is difficult due to a limited understanding of the viral replicative cycle. Utilizing a virus-like particle system, we have established roles for each of the viral structural components in assembly, release, and virus infectivity. The envelope glycoprotein, Gn, was discovered to be necessary and sufficient for packaging of the genome, nucleocapsid protein and the RNA-dependent RNA polymerase into virus particles. Additionally, packaging of the genome was found to be necessary for the efficient release of particles, revealing a novel mechanism for the efficient generation of infectious virus. Our results identify possible conserved targets for development of anti-phlebovirus therapies

Gn packages N.

<p><b>A.</b> BSR-T7/5 cells were transfected with genome and all of the structural proteins (WT), or one or more of the components was replaced with an equivalent amount of empty vector, (-Gn/-Gc, -Gn, or -Gc) or a plasmid expressing an allele of Gc that lacks the entire cytoplasmic tail (GcW1). Transfected cells were analyzed for protein expression by immunoblot. RVF-VLPs were immune precipitated from the clarified media from transfected cells and analyzed by immunoblot. The numbers below the immunoblots indicate the ratio of glycoprotein to N signal. <b>B</b>. BSR-T7/5 cells were transfected with the indicated plasmids and proteins were cross-linked at 48 h post-transfection. Mouse monoclonal anti-Gn antibodies were used to immune-precipitate Gn containing complexes. The cross-links were cleaved and then Gn-containing complexes were identified by immunoblot using rabbit anti-Gn or rabbit anti-N antibodies.</p

RVFV and RVF-VLPs have similar protein composition.

<p>Lysates from transfected (RVF-VLP) or RVFV-infected cells (C), and pelleted particles (P) were analyzed by immunoblot. The numbers below the immunoblots indicate the ratio of glycoprotein to N signal.</p

Gn recruits RdRp.

<p><b>A.</b> BSR-T7/5 cells were transfected with pRdRp or pGn, and the proteins were visualized with anti-RdRp and anti-Gn, respectively (green channel). Cellular resident Golgi apparatus proteins, GS-28 or β-COP were also labeled (red channel). Percentage of cells displaying co-localization of viral proteins with resident Golgi proteins is indicated with the number of cells counted in parentheses. <b>B.</b> BSR-T7/5 cells were transfected with pRdRp and either pGn/pGc, pGc, pGn, or pGnK48. Cells were incubated with anti-RdRp (green channel) and anti-Gn or anti-Gc (red channel), and then analyzed by immunofluorescence microscopy. Percentage of cells displaying co-localization of RdRp with Gn or Gc is indicated with the number of cells counted in parentheses.</p

Viral components required for efficient RVF-VLP release.

<p>BSR-T7/5 cells were transfected with genome and all of the structural proteins (WT), or one or more of the components was replaced with an equivalent amount of empty vector (-RNPCs, -Gn/Gc, -N, -genome, -RdRp). RNPCs refer to ribonucleoprotein complexes and are defined as genome, N, and RdRp. Transfected cells were analyzed for protein expression by immunoblot. RVF-VLPs were immune precipitated from the clarified media from transfected cells and analyzed by immunoblot. The numbers below the immunoblots indicate the ratio of glycoprotein to N signal.</p

Gc cytosolic tail is dispensable for infectivity.

<p>*Values are significantly different from -Gn/-Gc, p<0.01.</p

Encapsidated genome required for efficient cellular release.

<p>*Values are significantly different from WT RVF-VLP release efficiency, p<0.005.</p><p>†Value is significantly different from WT RVF-VLPs release efficiency, p<0.05.</p><p>ΔYields were below the limit of detection.</p><p>‡Insufficient number of replicates for std. dev. and p-value calculations.</p

RVFV and RVF-VLPs have similar morphology.

<p>RVFV and RVF-VLPs were harvested by ultracentrifugation and analyzed by transmission electron microscopy with negative staining. The particle sizes were measured, and the values listed are the mean sizes of particles with standard deviation.</p