Diagnosis and surgical treatment of an intracranial cyst in an alpaca cria.

CASE DESCRIPTION: A 7-day-old female alpaca was examined because of an acute onset of diffuse central neurologic deficits. CLINICAL FINDINGS: Diagnostic imaging with CT and MRI identified an intracranial cyst occupying approximately one-third to one-half of the dorsal portion of the cranial cavity, markedly displacing the cerebral hemispheres bilaterally. TREATMENT AND OUTCOME: Initial surgical management via trephination and needle drainage was only transiently effective at resolving the neurologic signs. Craniotomy and drainage and removal of the cyst lining resulted in a sustained improvement in neurologic status, and the cria remained clinically normal and well grown at follow-up 5 months after surgery. CLINICAL RELEVANCE: This report represented the first description of the successful treatment of an intracranial cyst in a New World camelid.

Characterization of equine P-selectin glycoprotein ligand-1 by using a specific monoclonal antibody.

P-selectin glycoprotein ligand (PSGL-1) is a widely distributed adhesion molecule that plays a critical role in regulating lymphocyte homing and leukocyte trafficking during inflammation. The lack of specific reagents for equine PSGL-1 (ePSGL-1) has prevented mechanistic studies regarding its function and regulation in the horse. We synthesized a ePSGL-1 peptide to generate a monoclonal antibody (mAb), ePL1. Using flow cytometry and Western blot, we showed that ePL1 binds specifically to ePSGL-1 in transfected mammalian cells. We also demonstrated that ePL1 binds to equine leukocytes and recognized a protein with molecular weight 165 and 280kDa under reducing and non-reducing condition, respectively, likely corresponding to ePSGL-1. Seventy percent of equine monocytes bound by both ePL1 and HECA-452, an antibody defining sLex-like carbohydrate epitope. Both ePL1 and HECA-452 recognized ePSGL-1 protein precipitated by equine P-selectin-IgG chimera. Neuraminidase treatment increased ePL1 binding and the molecular weight of ePSGL-1, O-sialoglycoprotein endopeptidase digestion and tyrosine mutation abolished ePL1 staining and recognition. The ePL1 specific binding epitope appears to be the polypeptide backbone of ePSGL-1 in the presence of tyrosine but the process is independent of sialylation modification. In conclusion, we provide evidence that this antibody can be used for cell surface staining and immune-blot analyses.

Hepatocellular carcinoma in the woodchuck model of hepatitis B virus infection.

The Eastern woodchuck ( Marmota monax ) harbors a DNA virus (Woodchuck hepatitis virus [WHV]) that is similar in structure and replicative life cycle to the human hepatitis B virus (HBV). Like HBV, WHV infects the liver and can cause acute and chronic hepatitis. Furthermore, chronic WHV infection in woodchucks usually leads to development of hepatocellular carcinoma (HCC) within the first 2-4 years of life. The woodchuck model has been important in the preclinical evaluation of safety and efficacy of the antiviral drugs now in use for treatment of HBV infection and continues to serve as an important, predictive model for innovative forms of therapy of hepatitis B using antiviral nucleosides and immune response modifiers alone or in combination. Almost all woodchucks that become chronic WHV carriers after experimental neonatal inoculation develop HCC with a median HCC-free survival of 24 months and a median life expectancy of 30-32 months. The woodchuck model of viral-induced HCC has been used effectively for the development of new imaging agents for enhancement of detection of hepatic neoplasms by ultrasound and magnetic resonance imaging. The chemoprevention of HCC using long-term antiviral nucleoside therapy has been shown in the woodchuck, and "proof of principal" has been established for some of the innovative, molecular methods for treatment of HCC. The model is available for fundamental investigations of the viral and molecular mechanisms responsible for hepatocarcinogenesis and should have substantial value for future development of innovative methods for chemoprevention and gene therapy of human HCC.

Evaluation of cytotoxicity and antiviral activity of recombinant human interferon alfa-2a and recombinant human interferon alfa-B/D hybrid against bovine viral diarrhea virus, infectious bovine rhinotracheitis virus, and vesicular stomatitis virus in vitro.

OBJECTIVE: To evaluate cytotoxicity and antiviral activity of recombinant human interferon alfa-2a and recombinant human interferon alfa-B/D hybrid against cytopathic and noncytopathic bovine viral diarrhea virus (BVDV), infectious bovine rhinotracheitis virus (IBRV), and vesicular stomatitis virus (VSV) in vitro. SAMPLE POPULATION: Primary bovine testicular cells and Mardin Darby bovine kidney cells. PROCEDURES: To evaluate cytotoxicity, cells were added to serial dilutions of each interferon. To evaluate antiviral activity of each interferon, interferons were serially diluted 1:10, and tissue culture cells were added; virus was then added at 3 time points. Prevention of viral infection by interferon was defined as failure to induce cytopathologic effect for VSV, IBRV, and cytopathic BVDV and failure to detect virus immunohistochemically for cytopathic and noncytopathic BVDV. RESULTS: No evidence of cytotoxicity in either cell line was detected after incubation with interferon alfa-2a or interferon alfa-B/D. However, reduced growth rates of tissue culture cells were detected for each interferon when undiluted interferon was tested. Comparable and profound antiviral activities against cytopathic and noncytopathic BVDV were evident for each interferon. Interferon alfa-2a and interferon a-B/D had comparable antiviral activities against VSV. Neither interferon had antiviral activity against IBRV. CONCLUSIONS AND CLINICAL RELEVANCE: The safety and marked in vitro antiviral activity against noncytopathic BVDV, cytopathic BVDV, and VSV suggest that interferons alfa-2a and alfa-B/D may be useful for treatment of natural disease after infection with these viruses.

Evaluation of antiviral activity and toxicity of recombinant human interferon alfa-2a in calves persistently infected with type 1 bovine viral diarrhea virus.

OBJECTIVE: To evaluate antiviral activity and toxicity of recombinant human interferon alfa-2a in calves persistently infected with noncytopathic type 1 bovine viral diarrhea virus (BVDV). ANIMALS: 5 Holstein heifers, 4 to 12 months of age. PROCEDURES: Calves persistently infected with noncytopathic type 1 BVDV were treated with recombinant human interferon alfa-2a every other day for 12 weeks. Viral loads were measured during the treatment period and compared with pre- and post-treatment values. Complete physical examinations were performed weekly, and calves were observed daily for signs of systemic illness. Complete blood counts and serum biochemical analyses were performed before, during, and after the treatment period. Because calves developed anemia during the treatment period, bone marrow biopsy specimens were collected. Antirecombinant human interferon alfa-2a antibody concentrations in serum samples obtained before, during, and after the treatment period were measured by use of an ELISA. RESULTS: Recombinant human interferon alfa-2a had no antiviral activity against noncytopathic type 1 BVDV in persistently infected calves. All calves developed microcytic anemia during the treatment period that persisted for up to 13 weeks after cessation of treatment. Anti-interferon antibodies were detected during the treatment period and persisted for at least 2 weeks after cessation of treatment. CONCLUSIONS AND CLINICAL RELEVANCE: Because of lack of in vivo antiviral activity against BVDV, recombinant human interferon alfa-2a has little promise as a therapeutic agent for the treatment of BVDV infection, at least in persistently infected cattle. Furthermore, treatment was associated with adverse immunologic and hematologic effects.