Hemorrhagic Herpes Simplex Virus Type 1 Nephritis: An Unusual Cause of Acute Allograft Dysfunction.

Our laboratory specializes in two areas, infectious diseases, particularlyviral diseases, and ultrastructure-function relationships. Electronmicroscopy (EM) is the focus of the investigative techniques and includes preparative methods such as negative staining, thin sectioning, ultracryomicrotomy and immunolabeling of acrylic and frozen sections.

We are especially interested in methods for diagnosing viral illnesses by EM, and are involved in developing better, more sensitive and faster,methods for detection. While molecular techniques for detecting organisms
are very sensitive, they all require specific reagents, and if the correct probe is not determined a priori, the test is negative. EM offers an open view of any viruses or unsuspected organisms that may be present. We make use of concentration and enhancement methods to increase the chances of
detecting viral agents in fluid specimens. Additionally, we have described a method for selecting small focal areas of pathology in tissue by confocal microscopy to be embedded and examined by EM, increasing the
chances of visualizing organisms. Infectious diseases are the leading cause of death worldwide and the third leading cause in the US. With advanced therapies for cancer patients and many patients living longer
with their disease, a whole new population of infectious disease-susceptible patients has emerged. Chemotherapy, radiation, and bone marrow transplantation are permitting longer survival, but cause
immunosuppression and consequently, strange, unusual diseases, such as polyomavirus infections, sometimes in uncommon body sites. We work closely with physicians to detect and monitor the clearance of
polyomavirus infections in bone marrow and kidney transplant patients. We detect food-borne outbreaks on campus, and we test numerous specimens from patients with infectious diseases. We also serve on the Duke Biodefense Team due to our capability to detect and differentiate poxvirus infections
from those of herpesvirus infections rapidly (within minutes).

Several research collaborations are underway. We have worked with Dr. David Pickup on a structural protein that directs intracellular virus particle movement and maturation. A project with Drs. William Parker and
Randal Bollinger, involves looking at microbes and mucous membrane immunity. It concentrates on biofilms in appendix and lower intestine. We are collaborating with Dr. Meta Kuehn on immunostaining bacterial
vesicles possibly containing endotoxin that have been internalized by
human cells. A different project with Drs. Celia LeBranche and Brian Cullen has examined morphological differences in various retrovirus outer membranes. With Dr. Barton Haynes' laboratory, we determined that cells transfected with single retroviral genes produced subviral particles. With Dr. Michael Hauser's lab, we are examining the difference of myotilin concentration in normal muscle and muscle from muscular dystrophy patients. We worked with a postdoctoral student in the laboratory of Dr. Shirish Shinolokar on staining and examining actin and actin-bundling protein by EM. Finally, we train and assist graduate students, post doctoral students and medical residents how to use electron microscopic techniques in their own studies.

RESEARCH ABSTRACT

Morphology and pathogenesis of Pneumocystis carinii, rapid viral diagnosis, particularly in the immunocompromised host.

Most of my research efforts are of an applied nature which involves adapting new methods of rapid viral diagnosis and testing those newer approaches against standard virological procedures. Validated new tests are incorporated into the standard work-up of appropriate virological specimens. Ultimately the expanded offerings result in a chargeable test for the laboratory. For example as monoclonal antibodies have become available for common respiratory viruses causing lower respiratory tract disease, we have expanded our Respiratory Viral Battery FA and perform that routinely on appropriate specimens as opposed to only performing a RSV FA. We also now charge for Toluidine Blue O staining for Pneumocystis carinii, which I modified and standardized for our lab.

I am also doing an ongoing epidemiological study of adenovirus subtypes in cases of disseminated infection in SCIDs patients. Some of the typing was done by Molly Rivadeneira, M.D. when she rotated through our laboratory. We now have 12 patients in this study.

Also I am studying the morphology and staining characteristics of Pneumocystis carinii when exposed to Calcofluor, a fluorescent brightener that stains cellulose and chitin. In addition to examining laboratory strains, isolates of P. carinii from DUMC patient material is currently being studied. We now have 14 patient isolates in this study.

Clinical research related to infectious complications of solid organ and bone marrow transplantation, with a particular interest in the treatment and rapid diagnosis of fungal disease. Training the next generation of Transplant Infectious Disease Physicians is a special focus of mine as the Principal Investigator of our Interdisciplinary T32 Training Program funded the NIH.