The safety of high-dose buprenorphine administered subcutaneously in cats.

The safety of a proprietary formulation of buprenorphine hydrochloride administered subcutaneously (SC) to young cats was investigated in a blinded, randomized study. Four cohorts of eight cats aged approximately 4 months were administered saline, 0.24, 0.72 or 1.20 mg/kg/day buprenorphine SC for nine consecutive days, representing 0×, 1×, 3× and 5× of the intended dose. Cats were monitored daily for evidence of clinical reactions, food and water intake and adverse events (AEs). Physical examinations, clinical pathology, vital signs and electrocardiograms (ECGs) were evaluated at protocol-specified time points. Complete necropsy and histopathologic examinations were performed following humane euthanasia. Four buprenorphine-treated cats experienced AEs during the study, two unrelated and two related to study drug administration. The two cats with AEs considered related to drug administration had clinical signs of hyperactivity, difficulty in handling, disorientation, agitation and dilated pupils in one 0.24 mg/kg/day cat and one 0.72 mg/kg/day cat. All of these clinical signs were observed simultaneously. There were no drug-related effects on survival, injection response, injection site inspections, body weight, food or water consumption, bleeding time, urinalysis, respiration rate, heart rate, ECGs, blood pressures, body temperatures, macroscopic examinations or organ weights. Once daily buprenorphine s.c. injections at doses of 0.24, 0.72 and 1.20 mg/kg/day for 9 consecutive days were well tolerated in young domestic cats.

γ134.5-deleted HSV-1-expressing human cytomegalovirus IRS1 gene kills human glioblastoma cells as efficiently as wild-type HSV-1 in normoxia or hypoxia.

Pathophysiological hypoxia, which fosters the glioma stem-like cell (GSC) phenotype, is present in high-grade gliomas and has been linked to tumor development, invasiveness and resistance to chemotherapy and radiation. Oncolytic virotherapy with engineered herpes simplex virus-1 (HSV-1) is a promising therapy for glioblastoma; however, the efficacy of γ(1)34.5-deleted HSVs, which have been used in clinical trials, was diminished in hypoxia. We investigated the ability of a chimeric human cytolomegalovirus (HCMV)/HSV-1 virus, which expresses the human CMV protein kinase R evasion gene IRS1 and is in preparation for clinical trials, to infect and kill adult and pediatric patient-derived glioblastoma xenografts in hypoxia and normoxia. Infectivity, cytotoxicity and viral recovery were significantly greater with the chimeric virus compared with the γ(1)34.5-deleted virus, regardless of oxygen tension. The chimeric virus infected and killed CD133+ GSCs similarly to wild-type HSV-1. Increased activation of mitogen-activated protein kinase p38 and its substrate heat-shock protein 27 (Hsp27) was seen after viral infection in normoxia compared with hypoxia. Hsp27 knockdown or p38 inhibition reduced virus recovery, indicating that the p38 pathway has a role in the reduced efficacy of the γ(1)34.5-deleted virus in hypoxia. Taken together, these findings demonstrate that chimeric HCMV/HSV-1 efficiently targets both CD133+ GSCs and glioma cells in hypoxia.

Laminar development of the mouse barrel cortex: effects of neurotoxins against monoamines.

The rodent somatosensory cortex is characterized by a unique cellular organization in the field of representation of the whiskers, called the barrelfield, which develops in layer IV during the 1st postnatal week in parallel with the establishment of the thalamo-cortical connections. This area is transiently densely innervated by serotonergic afferents during this period. Serotonin depletion delays the formation of barrels in the rat somatosensory cortex. However, no information is available to date on the time-course of the laminar differentiation of the cortex after monoaminergic depletion and the relative contribution of different monoaminergic inputs to this process. To address these issues, newborn mice were treated with selective neurotoxins (6-hydroxydopamine or 5,7-dihydroxytryptamine) at birth to destroy the catecholaminergic and monoaminergic cortical innervation, respectively. The parietal cortex of these animals was examined in Nissl-stained coronal sections prepared on different days of postnatal development (between P2 and P30). Compared with the controls, delayed growth and differentiation of the cortical layers II-IV were observed in the treated animals, most prominently between P2 and P16. From the 3rd postnatal week, no cytoarchitectonic difference could be detected. Although neonatal depletion of the cortical monoaminergic innervation does not affect the laminar organization of the adult mouse barrelfield, it significantly delays the time-course of development of several cortical layers. This delay generates a mismatch in the degree of maturation between cortical neurons and their afferents at a time when neuronal interactions are critical for the establishment of local circuitry.

Monoaminergic afferents to cortex modulate structural plasticity in the barrelfield of the mouse.

Electrolytic lesions of the follicles of a set of mystacial vibrissae, and their innervation, of the mouse placed during the early postnatal period result in a modification in appearance of the corresponding and of adjacent barrels in the somatosensory cortex of the adult animal. These changes can be evoked during the first 6 days of postnatal life--the so-called critical period. The pattern of these modifications varies with the age of the animal at which the lesion was placed. In order to evaluate the contribution of the monoaminergic cortical input to this type of plasticity, the noradrenergic and/or serotonergic afferents to the cerebral cortex of newborn mice were destroyed by systemic administration of various selective neurotoxic drugs (6-hydroxydopamine, 5,7-dihydroxytryptamine, N-(2-chloroethyl)-N-ethyl-2-bromobenzylamine). The animals were then subjected, on postnatal day 3 (P3; P0 = day of birth), to a lesion of the follicles of the large, caudal mystacial vibrissae of row C. Control animals were injected with vehicle solution only but had the same follicles lesioned. Compared with animals with intact monoaminergic afferents, those treated with neurotoxins showed a different changed barrel pattern, i.e. one that corresponded to a pattern normally obtained after a lesion placed at an earlier stage of development, i.e. at P2 or P1. Thus, monoaminergic depletion of the cortex results in a retardation of the maturation of the parietal cortex as defined by its plastic response to peripheral nerve injury.(ABSTRACT TRUNCATED AT 250 WORDS)