DNA vaccines encoding viral glycoproteins induce nonspecific immunity and Mx protein synthesis in fish.

Protective immunity by vaccination with plasmid DNA encoding a viral glycoprotein (G) has long been assumed to result from the induction of a specific immune response. We report here that the initial protection may be due to the induction of alpha/beta interferon, with long-term protection due to a specific response to the encoded viral G. DNA vaccines encoding the Gs of three serologically unrelated fish rhabdoviruses were used to vaccinate rainbow trout against a lethal challenge with infectious hematopoietic necrosis virus (IHNV). All three vaccines, each encoding the G gene of either IHNV (IHNV-G), snakehead rhabdovirus (SHRV) (SHRV-G), or spring viremia of carp virus (SVCV) (SVCV-G), elicited protective immunity against IHNV. Vaccinated fish were challenged at 30 or 70 days postvaccination with lethal doses of IHNV. At 30 days postvaccination, only 5% of fish that had received any of the G vaccines died, whereas more than 50% of the control fish succumbed to virus challenge. When fish were vaccinated and challenged at 70 days postvaccination, only 12% of the IHNV-G-vaccinated fish died compared to 68% for the SHRV-G- and 76% for the SVCV-G-vaccinated fish. Assays for trout Mx protein, an indicator of alpha/beta interferon induction, showed that only fish vaccinated with a G-containing plasmid produced high levels of Mx protein in the kidneys and liver. Interestingly, at day 7 after virus challenge, all of the fish vaccinated with the IHNV-G plasmid were negative for Mx, but the SHRV-G- and SVCV-G-vaccinated fish still showed detectable levels of Mx. These results suggest that DNA vaccines in fish induce an early, nonspecific antiviral protection mediated by an alpha/beta interferon and, later, a specific immune response.

Production of recombinant snakehead rhabdovirus: the NV protein is not required for viral replication.

Snakehead rhabdovirus (SHRV) affects warm water fish in Southeast Asia and belongs to the genus Novirhabdovirus by virtue of its nonvirion gene (NV). Because SHRV grows best at temperatures between 28 and 31 degrees C, we were able to use the T7 expression system to produce viable recombinant SHRV from a cloned cDNA copy of the viral genome. Expression of a positive-strand RNA copy of the 11, 550-nucleotide SHRV genome along with the viral nucleocapsid (N), phosphoprotein (P), and polymerase (L) proteins resulted in the generation of infectious SHRV in cells preinfected with a vaccinia virus vector for T7 polymerase expression. Recombinant virus production was verified by detection of a unique restriction site engineered into the SHRV genome between the NV and L genes. Since we were now able to begin examining the function of the NV gene, we constructed a recombinant virus containing a nonsense mutation located 22 codons into the coding sequence of the NV protein. The NV knockout virus was produced at a concentration as high as that of wild-type virus in cultured fish cells, and the resulting virions appeared to be identical to the wild-type virions in electron micrographs. These initial studies suggest that NV has no critical function in SHRV replication in cultured fish cells.

Interaction of steroidal alkaloid toxins with calcium channels in neuronal cell lines.

Depolarization with 50 mM K+ increased 45Ca2+ uptake into neuronal clonal cell lines NG108-15, N1E-115 and NH15-CA2. In each cell line this depolarization-induced uptake was blocked by inorganic and organic blockers of voltage sensitive calcium channels. However, tetrodotoxin (10(-6) M) was ineffective. Moreover, in the presence of tetrodotoxin, neither batrachotoxin nor veratridine inhibited the depolarization-induced uptake. The novel dihydropyridine BAY K8644 enhanced depolarization-induced 45Ca2+ uptake into each cell line in a nitrendipine reversible fashion. In the presence of tetrodotoxin, the BAY K8644/50 mM K+ stimulated uptake could be partially inhibited by batrachotoxin (10(-6) M) and veratridine (5 X 10(-5) M). These effects were not altered by the presence of scorpion venom (1 microgram/ml). The results indicate that both batrachotoxin and veratridine can modulate the effects of dihydropyridines on the gating properties of voltage sensitive calcium channels.

The gluteus maximus myocutaneous flap in the management of pressure sores.

The gluteus maximus myocutaneous flap is modified to obviate the necessity of a skin graft to the donor defect. This is a sufficiently large flap which, in the event of a recurrence, can be rerotated. The design and surgical technique of this flap for ischial and sacral pressure sores are described. This flap was used for 36 pressure sore without loss of any portion of the flap.

Alternatives to subcutaneous mastectomy.

It is our belief that subcutaneous mastectomy often leads to complications, with poor results. In addition, because a considerable amount of breast tissue may be left behind, its value as a prophylactic operation must be questioned. Two alternatives are presented. In our experience, these lead to fewer complications with a more thorough removal of the mammary gland, and achieve a better appearance. The alternatives are (1) simple mastectomy with secondary reconstruction; (2) the type of mastectomy described by Spira--with primary reconstruction using a subpectoral silicone prosthesis covered by muscle and an inferiorly-based dermal flap.

A filing system for transparencies.

A system has been developed to facilitate filing of 2 inches X 2 inches Kodachrome slides. This is coordinated with a diagnostic and therapeutic index that allows for easy retrieval. Compact metal filing cabinets are used, eliminating the need for subsequent rearrangement of slides. The system also provides a method for recall of patients. Preoperative photographs are maintained in plastic folders and checked periodically until the postoperative photographs have been taken.

Mondor's disease. An unusual complication of mammaplasty.

We present 6 cases of Mondor's disease which developed after plastic surgical operations on the breasts. All signs of the disease disappeared spontaneously after 3 or 4 months.

Experiences with the Pitangy method of correction of trochanteric lipodystrophy.

The operation for correction of trochanteric lipodystrophy has given good results in properly selected cases. It is a major procedure, however-one which has a significant postoperative morbidity.