Polyethylene glycol (PEG)ylated (stealth) immunoliposomes directed against human gliofibrillary acidic protein (GFAP) were prepared by coupling the thiolated monoclonal anti-GFAP antibodies with a maleimide derivative of phosphatidyl ethanolamine of the liposomal membrane. Experiments with cell cultures demonstrated specific and competitive binding of these immunoliposomes to embryonic rat brain astrocytes. Administered intravenously into rats, the immunoliposomes displayed typical kinetics with elimination half-lives of 8-15 hr. Being incapable of penetrating the unimpaired blood-brain barrier (BBB), these immunoliposomes, nevertheless, may be useful in delivering drugs to glial brain tumors (which continue to express GFAP) or to other pathological loci in the brain with a partially disintegrated BBB.

No abstract

Female BALB/c mice were subcutaneously immunized with recombinant VEGF-164. After 3 immunization cycles, splenic B cells from immunized mouse were fused with immortalized myeloma culture SP2/0-Ag14 cells. Screening of hybrid cells producing anti-VEGF antibodies was performed by ELISA and immunocytochemical analysis on cultured C6 glioma cells. Subsequent cloning yielded hybridoma stably expressing monoclonal anti-VEGF antibodies recognizing recombinant and native VEGF.

Methods of GFAP purification and obtaining of hybridoma cells producing monoclonal anti-GFAP antibodies and properties of GFAP preparation were described. The immunobloting data on specificity of obtained monoclonal antibodies are presented. A new method of GFAP immunoenzyme assay based on GFAP preparation and anti-GFAP antibodies was elaborated. Standardization of the immunoenzyme system was shown in tests for specificity, accuracy, and reproducibility.

Sir: Pathological crying after stroke is characterized by outbursts of weeping and tearfulness that are not related to an underlying emotional state, but that can be very distressing to the patient.1 Disturbed serotonergic transmission is believed to play a role.2 A number of treatments have been reported as successful, including sertraline, 3 amitriptyline, 9 and nortriptyline. 10 We report the first case of venlafaxine used to successfully treat poststroke pathological crying.Case report. Mr. A, a 67-year-old right-handed man, suffered a hypertensive stroke in 1999 manifested at first by slurred speech and collapse. Computed tomography scan of the head revealed hemorrhage in the left parietal lobe and basal ganglia, compressing the left lateral ventricle, with minimal edema and no midline shift. Also noted were old, small rightsided infarcts with no apparent clinical sequelae.Mr. A was transferred to the rehabilitation service approximately 2 weeks after his stroke. We were asked to evaluate him at that time for crying spells. He denied any past psychiatric history or past untreated mood symptoms. Since the stroke, he reported having between 5 and 20 crying spells daily that were not associated with any change in mood. He did report feeling distressed by these spells but did not endorse symptoms of depression. He denied episodes of pathological laughter. The Pathological Laughter and Crying Scale (PLACS) 10 was administered, on which the patient scored 14 points.After informed consent was obtained from the patient, venlafaxine was initiated at a dose of 37.5 mg twice daily. Within 24 hours, the patient reported complete resolution of his crying spells. Follow-up at 2 weeks revealed a score of 0 on the PLACS. We continued to see him at the rehabilitation service and during his stay in the nursing home care unit. Throughout that 6-month period, he remained symptom-free and did not require any increase in dose.Several tricyclic antidepressants and selective serotonin reuptake inhibitors, as well as other medications, have been reported to dramatically reduce the frequency and severity of pathological crying spells after stroke. Venlafaxine is a serotonin-norepinephrine reuptake inhibitor that may be particularly useful in older patients due to its relatively low protein binding, low anticholinergic effects, low sedation, low orthostatic hypotension, and relatively favorable cytochrome P450 profile. 11,12 While there is the possibility of hypertension with doses over 300 mg daily, 11 which would be a concern in a stroke patient, we found that a low dose of venlafaxine was effective in providing complete relief from this often distressing and embarrassing sequela of stroke.We considered the possibility that since the patient had a hemorrhagic stroke, he might have had seizures manifested by crying. It is known that seizures occur more commonly with hemorrhagic stroke than with ischemic stroke, particularly when the stroke is cortical in location.13 Additionally, we know that venlafaxine acts to increase catechol...