Celia R. Valverde

Comparisons of the intraocular tissue distribution, pharmacokinetics, and safety of 125I-labeled full-length and fab antibodies in rhesus monkeys following intravitreal administration

Access of recombinant proteins to the retina following intravitreal administration is poorly understood. A study was conducted in male Rhesus monkeys (15 to 28 mo of age; 2.8-3.3 kg) in order to compare the intraocular tissue distribution, pharmacokinetics, and safety of 125Iodine (I)-labeled full-length humanized rhuMAb HER2 antibody (148 kD) and of 125I-labeled humanized rhuMAb vascular endothelial growth factor Fab antibody (48.3 kD) following bilateral bolus intravitreal injection on day 0 (5 animals/group). The dose administered to each eye was 25 μg (9-10 μCi) in 50 μl. Animals were euthanatized on day 0 (1 hr postdose) and on days 1, 4, 7, and 14. Safety assessment included direct ophthalmoscopy, intraocular pressure measurements, clinical observations, body weight, and hematology and clinical chemistry panels. Blood and vitreous samples were collected daily (blood only) and at necropsy for pharmacokinetics and analysis for antibodies to the test materials; the ocular tissue distribution of the test material was evaluated by microautoradiography. All animals completed the study. Microautoradiography demonstrated that the full-length antibody did not penetrate the inner limiting membrane of the retina at any of the time points examined. In contrast, the Fab antibody fragment diffused through the neural retina to the retinal pigment epithelial layer at the 1-hr time point and persisted in this location for up to 7 days. Systemic exposure to test material was low but variable: the highest plasma concentration of the full-length antibody was 20.3 ng/ml, whereas plasma concentrations for the Fab antibody remained below the limit of quantitation (i.e., <7.8 ng/ml). An immune response to the test material was not evident in either treatment group. The half-life in vitreous was 5.6 days for the full-length antibody and 3.2 days for the Fab antibody. The shorter intravitreal half-life of the Fab antibody is related to its smaller size and its significant diffusion through the retinal layers. The differences in pharmacokinetics and tissue distribution that are noted between the full-length and Fab antibodies in this study identify potential therapeutic approaches that may be exploited in specific disease conditions.

Passive Immunization of Newborn Rhesus Macaques Prevents Oral Simian Immunodeficiency Virus Infection

To determine if passively acquired antiviral antibodies modulate virus transmission and disease progression in human pediatric AIDS, the potential of pre- and postexposure passive immunization with hyperimmune serum to prevent oral simian immunodeficiency virus (SIV) infection or disease progression in newborn rhesus macaques was tested. Untreated neonates became infected after oral SIV inoculation and had high viremia, and most animals developed fatal AIDS within 3 months. In contrast, SIV hyperimmune serum given subcutaneously prior to oral SIV inoculation protected 6 newborns against infection. When this SIV hyperimmune serum was given to 3 newborns 3 weeks after oral SIV inoculation, viremia was not reduced, and all 3 infants died within 3 months of age due to AIDS and immune-complex disease. These results suggest that passively acquired antihuman immunodeficiency virus (HIV) IgG may decrease perinatal HIV transmission. However, anti-HIV IgG may not impart therapeutic benefit to infants with established HIV infection.

Biological Effects of Short-Term or Prolonged Administration of 9-[2-(Phosphonomethoxy)Propyl]Adenine (Tenofovir) to Newborn and Infant Rhesus Macaques

ABSTRACT The reverse transcriptase inhibitor 9-[2-(phosphonomethoxy)propyl]adenine (PMPA; tenofovir) was previously found to offer strong prophylactic and therapeutic benefits in an infant macaque model of pediatric human immunodeficiency virus (HIV) infection. We now summarize the toxicity and safety of PMPA in these studies. When a range of PMPA doses (4 to 30 mg/kg of body weight administered subcutaneously once daily) was administered to 39 infant macaques for a short period of time (range, 1 day to 12 weeks), no adverse effects on their health or growth were observed; this included a subset of 12 animals which were monitored for more than 2 years. In contrast, daily administration of a high dose of PMPA (30 mg/kg subcutaneously) for prolonged periods of time (>8 to 21 months) to 13 animals resulted in a Fanconi-like syndrome (proximal renal tubular disorder) with glucosuria, aminoaciduria, hypophosphatemia, growth restriction, bone pathology (osteomalacia), and reduced clearance of PMPA. The adverse effects were reversible or were alleviated following either complete withdrawal of PMPA treatment or reduction of the daily regimen from 30 mg/kg to 2.5 to 10 mg/kg subcutaneously. Finally, to evaluate the safety of a prolonged low-dose treatment regimen, two newborn macaques were started on a 10-mg/kg/day subcutaneous regimen; these animals are healthy and have normal bone density and growth after 5 years of daily treatment. In conclusion, our findings suggest that chronic daily administration of a high dose of PMPA results in adverse effects on kidney and bone, while short-term administration of relatively high doses and prolonged low-dose administration are safe.

Autonomic Mediation of Glucagon Secretion During Insulin-Induced Hypoglycemia in Rhesus Monkeys

Autonomic activation mediates the majority of the increase of glucagon secretion during insulin-induced hypoglycemia in several species including dogs, mice, and rats. However, the role of the autonomic nervous system to increase glucagon during hypoglycemia in humans remains controversial, and investigations in nonhuman primates have not been previously conducted. The autonomic contribution to glucagon secretion during hypoglycemia in a nonhuman primate was examined by two independent pharmacological approaches. Glucagon responses to clamped insulin-induced hypoglycemia were compared in conscious rhesus monkeys in the presence or absence of ganglionic blockade with trimethaphan, or during combined muscarinic and adrenergic receptor blockade with atropine, propranolol, and tolazoline. Insulin- induced hypoglycemia (plasma glucose = 1.9 ± 0.1 mmol/l) activated parasympathetic nerves to the pancreas as assessed by increased plasma pancreatic polypeptide (PP) levels (δ= 135.0 ± 36.8 pmol/l, P < 0.01), produced sympathoadrenal activation as assessed by elevations of plasma epinephrine (EPI) (δ = 22.3 ± 2.95 nmol/l, P < 0.0005) and norepinephrine (NE) (δ = 3.72 ± 0.77 nmol/l, P < 0.0025) and increased plasma inununoreactive glucagon (IRG) (δ = 920 ± 294 ngfl, P < 0.025). Nicotinic ganglionic blockade with trimethaphan prevented parasympathetic (δPP = 16.5 ± 16.3 pmol/l, P < 0.01 vs. control) and sympathoadrenal (δEPI = 1.52 ± 0.98 nmol/l; δNE = -0.62 ± 0.24 nmol/l, both P < 0.0025 vs. control) activation during hypoglycemia and inhibited the IRG response by 70% (δ = 278 ± 67 ng/l,P < 0.025 vs. control). Combined muscarinic and adrenergic receptor blockade reduced parasympathetic activation (δPP = 48.3 ± 16.3 pmol/l, P < 0.01 vs. control) and inhibited the IRG response by a similar degree to ganglionic blockade (δIRG = 284 ± 60 ngfl, P < 0.025 vs. control). These results demonstrate by two independent pharmacological approaches that autonomic activation makes a substantial contribution to increased glucagon secretion during hypoglycemia of ∼2.0 mmol/l in a species of nonhuman primate.

Anesthetic management in feline renal transplantation

OBJECTIVE To document perioperative and anesthetic management of 30 feline renal transplant recipients (1996-1998). STUDY DESIGN Retrospective clinical study. ANIMALS Thirty adult cats in end-stage renal failure that underwent heterotopic renal transplantation. MATERIALS AND METHODS The medical records were reviewed from 30 feline heterotopic renal transplant recipients. Cases were included only if they had been treated for hypertension using a beta-adrenergic antagonist, a calcium channel blocker or hemodialysis. Data regarding signalment, preoperative management, surgical technique, type and doses of anesthetics administered, perioperative hemodynamics and intra- and postoperative complications, postoperative analgesia, morbidity and early mortality were recorded. Data were expressed as mean ± SD. RESULTS Preanesthetic medication included a combination of an anticholinergic and an opioid (oxymorphone). Anesthesia induction was performed mostly with isoflurane and oxygen delivered by mask. Anesthesia maintenance was primarily achieved with isoflurane in 100% oxygen. Nitrous oxide was often used as part of the anesthetic technique. The mean duration of anesthesia was 4.6 hours ± 27 minutes. The mean renal allograft ischemic time was 60 minutes. During the anesthetic period, the majority of the recipient cats received either fresh whole blood (FWB) (N = 25, 83%), cross-matched packed red blood cells (PRBC) (N = 3, 10%) or fresh frozen plasma (FFP) (N = 2, 7%) combined with a balanced electrolyte solution. Blood products administered averaged 63 ± 34 mL and crystalloid 94 ± 62 mL. The most common treated intraoperative complications were hypotension (N = 14, 47%), hypothermia (N = 13, 43%), metabolic acidosis (N = 11, 37%), hypocalcemia (N = 5, 17%), hypoglycemia (N = 4, 13%), hypertension (N = 2, 7%), bradycardia (N = 1, 3%), and ventricular premature contractions (N = 1, 3%). All cats received opioid analgesics postoperatively. Complications observed in the first 24 hours postoperatively were hypertension (N = 20, 67%), hematuria (N = 14, 47%), electrolyte disturbances (N = 9, 30%), temperature imbalances (N = 5, 17%), decreased PCV requiring blood transfusion (N = 5, 17%), decreased perfusion of a foot associated with external iliac anastomosis technique (N = 5, 17%), seizures associated with hypertension (N = 3, 10%), uroabdomen (N = 2, 7%), acute graft rejection (N = 1, 3%) and, corneal ulceration (N = 1, 3%). Survival rates in the perioperative period were 100, 96.7, and 93.4% intraoperatively, at 24 hours, and 7 days following surgery. CONCLUSION Successful anesthesia can be performed in critically ill renal transplant recipients. However, for optimal graft function and patient survival, normothermia, normovolemia, normotension, and normal acid-base and electrolyte balance should be carefully maintained. Successful anesthetic management requires understanding of the pathophysiology of end-stage renal disease and the maintenance of homeostasis during the different stages of the perioperative period.

CHRONIC OTITIS EXTERNA/MEDIA WITH TOTAL EAR CANAL ABLATION AND BULLA CURETTAGE IN A NORTH AMERICAN BISON (BISON BISON)

Abstract An adult female North American bison (Bison bison) with a chronic otitis externa/media of the right ear was examined because of a 4-mo history of intermittent anorexia, apparent painful behavior, and auricular discharge from the right ear. Computerized tomography (CT) demonstrated osteolysis of the tympanic, petrous, and squamous aspects of the temporal bone with soft tissue replacement and sclerosis of the right bulla. A total ear canal ablation with bulla curettage was performed, and cefazolin-impregnated polymethacrylate beads were left within the right bulla and the remnant temporal bone. Six months after the surgery, the bison had no clinical signs of otitis media.