Small-animal research imaging devices.

The scientific study of living animals may be dated to Aristotle's original dissections, but modern animal studies are perhaps a century in the making, and advanced animal imaging has emerged only during the past few decades. In vivo imaging now occupies a growing role in the scientific research paradigm. Imaging of small animals has been particularly useful to help understand human molecular biology and pathophysiology using rodents, especially using genetically engineered mice (GEM) with spontaneous diseases that closely mimic human diseases. Specific examples of GEM models of veterinary diseases exist, but in general, GEM for veterinary research has lagged behind human research applications. However, the development of spontaneous disease models from GEM may also hold potential for veterinary research. The imaging techniques most widely used in small-animal research are CT, PET, single-photon emission CT, MRI, and optical fluorescent and luminescent imaging.

A potential synergistic anticancer effect of paclitaxel and amifostine on endometrial cancer.

Although paclitaxel is one of the most effective chemotherapeutic agents, its usefulness is still limited in advanced and recurrent endometrial cancer. Amifostine protection of normal tissues against the side effects of chemotherapeutic agents has been clinically proven in cancer patients; however, its application in endometrial cancer has not been fully evaluated. We have investigated the use of paclitaxel and amifostine in controlling the growth of poorly differentiated endometrial cancer cells, Hec50co, in vitro and in vivo. Our studies show that amifostine had direct anticancer effects on endometrial cancer cells in vitro by arresting the cell cycle at the G1 phase and inducing apoptosis. Amifostine also inhibited s.c. tumor growth in athymic mice. Paclitaxel IC50 value was reduced from 14 to 2 nmol/L with pretreatment of a single dose of 178 micromol/L of amifostine for 72 hours. Amifostine also synergized with paclitaxel in the arrest of the cell cycle at the G2-M phase and in the induction of apoptosis. This two-drug regimen inhibited s.c. tumor growth as well as improved mouse survival significantly more than paclitaxel alone. Amifostine also significantly improved paclitaxel-induced cytotoxic effects on peripheral blood profiles. Our studies show that amifostine has direct anticancer effects on endometrial cancer. Our data have also shown a potential anticancer synergy between amifostine and paclitaxel in vitro and in vivo, whereas amifostine maintained a protective role in peripheral blood profiles. The dual specificity of amifostine action should be further investigated.

Age-related differences in amplification of covalently closed circular DNA at early times after duck hepatitis B virus infection of ducks.

Inoculation of 3-day-old (3D) or 3-week-old (3W) ducklings with duck hepatitis B virus results in chronic or transient infection, respectively. We previously showed that rapid production of neutralizing antibody following inoculation of 3W ducklings prevents virus from spreading in the liver and leads to a transient infection (Y.-Y. Zhang and J. Summers, J. Virol. 78:1195-1201, 2004). In this study we further investigated early events of viral infection in both 3D and 3W ducks. We present evidence that a lower level of virus replication in the hepatocytes of 3W birds is an additional factor that probably favors transient infection. We suggest that lower virus replication is due to a less rapid covalently closed circular DNA amplification, leading to lower viremias and a slower spread of infection in the liver, and that the slower spread of infection in 3W ducks makes the infection more sensitive to interruption by the host immune responses.

Selecting an external light source for fetal biophysical testing.

OBJECTIVE: To select the most intense light source that penetrates tissues and is safe for fetal biophysical testing STUDY DESIGN: A 3-step series of experiments was undertaken using a digital light meter (Extech Light Probe Meter, Extech Instruments, Waltham, Massachusetts). First, the density of light through a light filter was compared between the sun and 4 commercially available light sources. Second, penetration of light through the abdominal subcutaneous tissue (3-4 cm thick) of 6 pigs was compared between these light sources. Last, the skin reaction to the preferred light source and the distance in penetrating the uterine cavity were assessed in 50 pregnant women. RESULTS: Light emitted from a halogen bulb was significantly more intense than from a photographic flash, krypton bulb or penlight. Maximal intensity was gained with the light source placed against the skin. The halogen bulb penetrated a pig's abdominal wall more than the photographic flash or krypton bulb. The thickness of a pig's abdominal wall is similar to the distance in pregnant humans near term from the skin to the uterine cavity. No thermal injury or discomfort to the skin was observed for exposures <10 seconds. CONCLUSION: The halogen bulb was safe, penetrated tissues most effectively and was the best light source for fetal biophysical testing.

Smart intramedullary rod for correction of pediatric bone deformity: a preliminary study.

We were interested in determining if a smart intramedullary rod made of nitinol shape-memory alloy is capable of correcting deformed immature long bones. Because of limitations in our study design the process was reversed in that we examined the smart rod's ability to create a deformity rather than to correct one. Smart rods of different lengths and diameters were heat-treated to resume a radius of curvature of 30 to 110 mm. The low and high temperature phases of the smart rods were set, respectively, at 0 degrees C to 4 degrees C and 36 degrees C to 38 degrees C. The preshaped smart intramedullary rods were implanted in the cooled martensite phase in the medullary canal of the tibia in eight rabbits, where they restored their austenite form, causing a continuous bending force. On a weekly basis anteroposterior and lateral radiographs of the surgically treated tibia and the contralateral tibia were obtained for comparison. Rabbits were euthanized 6 weeks after surgery and computed tomography scans of both tibias were used for image analysis. Smart rods with a larger radius of curvature showed only minimal signs of remodeling; however, rods with a radius of curvature of 50 and 70 mm generated enough force history to create bone remodeling and deformation. The amount of bone deformation was highly magnified when unicortical corticotomy on the tension side was done. Based on this preliminary study the technology of the smart intramedullary rod may provide a valuable alternative method to correct pediatric skeletal deformities.

Single-cell analysis of covalently closed circular DNA copy numbers in a hepadnavirus-infected liver.

Hepatitis B virus (hepadnavirus) infections are maintained by the presence of a small and regulated number of episomal viral genomes [covalently closed circular DNA (cccDNA)] in the nuclei of infected cells. Although a number of studies have measured the mean copy number of cccDNA molecules in hepadnaviral-infected cells, the distribution of individual copy numbers have not been reported. Using a PCR-based assay, we examined the number of cccDNA molecules of the duck hepatitis B virus in single nuclei isolated from the liver of a chronically infected duck over the course of 131 days of infection. Nuclei were isolated from frozen serial biopsies and individually deposited into PCR microplates by flow sorting. Each nucleus was assayed by nested PCR for cccDNA and for cellular IFN-alpha genes as an internal control. We found that 90% of the nuclei assayed contained between 1 and 17 cccDNA molecules, with the remaining 10% containing more (90% confidence), and that differences in the mean number of copies and distribution of copy numbers occurred within the same animal at different times postinfection. Overall, the data suggest (i) that the number of cccDNA molecules per cell may fluctuate over time, and (ii) that, according to these fluctuations, a substantial fraction of cells may contain only one or a few copies. We infer from the results that infected hepatocytes express virus at different levels and that during cell division it is possible to segregate cells containing no cccDNA.