Ebola virus persistence and disease recrudescence in the brains of antibody-treated nonhuman primate survivors.

Effective therapeutics have been developed against acute Ebola virus disease (EVD) in both humans and experimentally infected nonhuman primates. However, the risk of viral persistence and associated disease recrudescence in survivors receiving these therapeutics remains unclear. In contrast to rhesus macaques that survived Ebola virus (EBOV) exposure in the absence of treatment, we discovered that EBOV, despite being cleared from all other organs, persisted in the brain ventricular system of rhesus macaque survivors that had received monoclonal antibody (mAb) treatment. In mAb-treated macaque survivors, EBOV persisted in macrophages infiltrating the brain ventricular system, including the choroid plexuses. This macrophage infiltration was accompanied by severe tissue damage, including ventriculitis, choroid plexitis, and meningoencephalitis. Specifically, choroid plexus endothelium-derived EBOV infection led to viral persistence in the macaque brain ventricular system. This resulted in apoptosis of ependymal cells, which constitute the blood-cerebrospinal fluid barrier of the choroid plexuses. Fatal brain-confined recrudescence of EBOV infection manifested as severe inflammation, local pathology, and widespread infection of the ventricular system and adjacent neuropil in some of the mAb-treated macaque survivors. This study highlights organ-specific EBOV persistence and fatal recrudescent disease in rhesus macaque survivors after therapeutic treatment and has implications for the long-term follow-up of human survivors of EVD.

Biological variation of thromboelastrography variables in 10 clinically healthy horses.

OBJECTIVE: To assess the utility of population-based reference intervals (PRIs) for interpreting thromboelastography (TEG) variables in horses using biological variation data. DESIGN: Prospective cohort biologic variation study conducted over a 5-week period. SETTING: Veterinary teaching hospital and research facility. ANIMALS: Ten clinically healthy horses randomly selected from a veterinary school research and teaching herd. INTERVENTIONS: Horse health was determined using physical examination, CBC, and biochemical and coagulation profiles prior to the start of the study. Subsequently, once weekly blood sampling for TEG testing was performed for 5 weeks. MEASUREMENTS AND MAIN RESULTS: The 4 TEG variables reaction time (R), clot formation time (K), angle, and maximum amplitude (MA) were measured, and coefficient of variation representing within- and between-horse biological variation (CVi and CVg , respectively) and coefficient of variation representing analytical variation (CVa ) were calculated using a nested ANOVA after removing outlier data. The CVi , CVg , and CVa for R were 26.8%, 5.2%, and 5.9%; for K were 31.0%, 0.0%, and 5.9%; for angle were 9.4%, 6.2%, and 21.7%; and for MA were 3.4%, 4.1%, and 4.4%, respectively. Index of individuality (IOI) was then calculated for each variable using the formula {( CVi² + CVa²/CVg²)}¹/². IOI for R was 5.3, for angle was 3.8, and for MA was 1.4; IOI was not assessed for K. CONCLUSIONS: PRIs are appropriate for TEG variables, R, angle, and MA when interpreting results from individual horses based on calculated IOI values equal to or greater than 1.4. PRIs are likely appropriate when interpreting K, but IOI could not be calculated for this variable.

Hypercalcemia and parathyroid hormone-related peptide expression in a dog with thyroid carcinoma and histiocytic sarcoma.

A 9.5-year-old, male castrated Walker Hound was presented for evaluation of progressive weakness, anorexia, and weight loss. Imaging revealed multiple abdominal and thoracic masses and ascites; fine-needle aspirates of mesenteric and splenic masses confirmed malignancy, most likely histiocytic sarcoma. Laboratory analyses revealed increased ionized calcium and parathyroid hormone-related peptide (PTH-rP) concentrations, and concurrent low-normal parathyroid hormone concentration, consistent with humoral hypercalcemia of malignancy. Necropsy was performed after euthanasia. The dog had disseminated histiocytic sarcoma, including sarcomatosis, as well as bilateral thyroid carcinoma. PTH-rP immunostaining was positive in the thyroid carcinoma but negative in the histiocytic neoplasm. These results suggest that thyroid carcinoma-associated hypercalcemia can be caused by tumor secretion of PTH-rP.

Identification of blue staining vaccine-derived material in inflammatory lesions using cultured canine macrophages.

BACKGROUND: Vaccine reactions are described in cytology textbooks as having eosinophilic to magenta colored globules within and admixed with inflammatory cells. Recently, we have seen increased numbers of inflammatory lesions containing blue to blue-gray globular material, with historical information suggesting an association with rabies vaccination. OBJECTIVES: The purpose of the study was to confirm the blue-gray and the eosinophilic material observed microscopically in some inflammatory lesions as being vaccine-derived. METHODS: Three different vaccines were cytocentrifuged and Wright stained. Vaccine aliquots were also added to the culture media of canine-derived macrophages for 24 hours and the cells subsequently harvested, cytocentrifuged, and Wright stained. The globular material present in both preparations was compared to that observed in vaccine-induced inflammatory lesions. Morin staining was used to identify metal within vaccine material in both in vitro- and in vivo-derived cytology samples. RESULTS: Vaccine-derived material has a characteristic color and appearance. Appearance of the material was consistent in cytologic samples, in cells incubated with the vaccine, and in cytocentrifuged preparations of the vaccine vial contents. The blue-gray globules stained positively for Morin stain, while the eosinophilic material did not stain. CONCLUSIONS: Vaccine-induced inflammatory lesions may contain blue to blue-gray or magenta stained globular material. Blue-gray material was associated with administration of rabies vaccine Imrab 3 TF and the observed material may be metal-containing adjuvant. Magenta material was associated with other vaccines and negative for Morin stain, suggesting a metal-free adjuvant.

Effects of freezing on tissue factor activity in a thromboelastography assay.

BACKGROUND: Human recombinant tissue factor (TF) can be used to activate viscoelastic coagulation assays. Although the package insert indicates that TF should not be frozen, published scientific data are not available. Ability to store frozen aliquots of TF would increase laboratory efficiency and decrease costs associated with performing TF-activated assays. OBJECTIVE: The objectives of this study were to determine the effects of freezing and storage time on TF's ability to activate commercially available quality control material in thromboelastography (TEG). METHODS: TF was diluted and frozen at -20°C and -70°C for 72 hours, one week, 2 weeks, 3 months, and 6 months, and used for TEG after thawing. TF activation of control material was also assessed after TF storage at room temperature for 0, 24, and 48 hours. Time to fibrin formation (R), rate of clot formation (K and angle α), and clot strength (MA) were measured, and ANOVA used to identify differences. RESULTS: There were no significant differences in mean α and MA regardless of storage time or temperature. Means for R were not significantly increased at any time point after storage at room temperature or -70°C, but significant increases in mean R were observed after storage at -20°C starting after one week and continuing up to 6 months. CONCLUSION: TF can be stored at room temperature for at least 48 hours, stored at -20°C for 72 hours, and stored at -70°C for up to 6 months without significant loss of activity for TEG.

The hallucinogen 1-[2,5-dimethoxy-4-iodophenyl]-2-aminopropane (DOI) increases cortical extracellular glutamate levels in rats.

Activation of the cerebral cortex is seen during hallucinations. The 5-HT(2A/C) agonist 1-[2,5-dimethoxy-4-iodophenyl]-2-aminopropane (DOI) is a potent hallucinogen that has been proposed to act by targeting 5-HT(2A) heteroceptors on thalamocortical neurons and eliciting release of glutamate from these cells, which in turn drives cortical neurons. We used in vivo microdialysis to determine if DOI increases extracellular glutamate levels. Systemic administration of DOI significantly increased extracellular glutamate levels in the somatosensory cortex of the freely-moving rat. Similarly, intracortical administration of DOI by reverse dialysis increased cortical extracellular glutamate levels. No consistent changes in either extracellular GABA or glycine levels were observed in response to DOI. The increase in glutamate levels elicited by intracortical DOI was blocked by treatment with the selective 5-HT(2A) antagonist MDL 100,907. These data are consistent with the hypothesis that 5-HT(2A) receptor-mediated regulation of glutamate release is the mechanism through which hallucinogens activate the cerebral cortex.