REVIEW ARTICLEThrombus formation: direct real-time observation and digital analysis of thrombus assembly in a living mouse by confocal and widefield intravital microscopy

Summary.  We have developed novel instrumentation using confocal and widefield microscopy to image and analyze thrombus formation in real time in the microcirculation of a living mouse. This system provides high-speed, near-simultaneous acquisition of images of multiple fluorescent probes and a brightfield channel, and supports laser-induced injury through the microscope optics. Although this imaging facility requires interface of multiple hardware components, the primary challenge in vascular imaging is careful experimental design and interpretation. This system has been used to localize tissue factor during thrombus formation, to observe defects in thrombus assembly in genetically altered mice, to study the kinetics of platelet activation and P-selectin expression following vascular injury, to analyze leukocyte rolling on arterial thrombi, to generate three-dimensional models of thrombi, and to analyze the effect of antithrombotic agents in vivo .     

Patient knowledge and expectations for functional recovery after stroke

OBJECTIVE: Understanding the causes and outcomes of stroke is important for stroke survivors and may affect their success in rehabilitation and their risk of recurrent stroke; therefore, this study was performed to assess the knowledge and expectation of functional recovery in stroke patients undergoing acute inpatient rehabilitation. DESIGN: Survey study of 50 consecutive stroke patients undergoing inpatient rehabilitation at a single urban rehabilitation hospital. RESULTS: Forty-six percent of participants were able to correctly identify whether they had sustained a cerebral infarct or hemorrhage. Rehabilitation length of stay was, on average, 1 wk longer than anticipated by patients. Patients overestimated their functional abilities on initial assessment and at discharge compared with staff assessments, with some improvement in accuracy for discharge predictions. Ninety-four percent of participants expected to be discharged home, and most achieved this goal. Although no participant anticipated discharge to a nursing home, 10% of patients were discharged to this location. CONCLUSIONS: Knowledge of stroke and its treatment was limited, and expectations for recovery tended to exceed actual accomplishments. There are significant areas of opportunity for enhanced educational efforts for stroke patients undergoing inpatient rehabilitation.     

Dehydroepiandrosterone and biologics in the treatment of systemic lupus erythematosus

Our understanding of the immune system in health and disease has greatly increased over this past decade. An outgrowth of this burgeoning knowledge has been the discovery of novel strategies to treat a variety of autoimmune diseases. As a result, significant advances have been made in the treatment of arthritis with a half-dozen new treatments approved between September, 1998 and November, 1999. Although no new therapies have been approved for the treatment of systemic lupus erythematosus (SLE), this decade has witnessed an unprecedented amount of activity in drug development for this disease. This article specifically addresses the current status of drug development in the area of hormonal and biologic therapies for SLE.     

LJP 394 for the prevention of renal flare in patients with systemic lupus erythematosus: results from a randomized,double-blind,placebo-controlled study

OBJECTIVE: To determine whether LJP 394 delays or prevents renal flare in patients with systemic lupus erythematosus (SLE) and a history of renal disease. METHODS: In a 76-week, double-blind, placebo-controlled study, 230 SLE patients were randomized to receive 16 weekly doses of 100 mg of LJP 394 or placebo, followed by alternating 8-week drug holidays and 12 weekly doses of 50 mg of LJP 394 or placebo. An assay measuring the affinity of the serum IgG fraction for the DNA epitope of LJP 394 identified a high-affinity population of patients (189 of 213 patients; 89% taking LJP 394 and 90% taking placebo). Analyses were performed on both the intent-to-treat population and the high-affinity population. RESULTS: Anti-double-stranded DNA antibodies decreased and C3 levels tended to increase during treatment with LJP 394. In the intent-to-treat population, the time to renal flare was not significantly different between treatment groups, but patients taking LJP 394 had a longer time to institution of high-dose corticosteroids and/or cyclophosphamide (HDCC) and required 41% fewer treatments with HDCC. In the high-affinity population, the LJP 394 group experienced a longer time to renal flare, 67% fewer renal flares, longer time to institution of HDCC, and 62% fewer HDCC treatments compared with the placebo group. In patients with serum creatinine levels >/=1.5 mg/dl at study entry, those taking LJP 394 had 50% fewer renal flares; no renal flares were observed in the high-affinity group taking LJP 394. Serious adverse events were observed in 25 of the 114 LJP 394-treated patients (21.9%) and 34 of the 116 placebo-treated patients (29.3%). CONCLUSION: Treatment with LJP 394 in patients with high-affinity antibodies to its DNA epitope prolonged the time to renal flare, decreased the number of renal flares, and required fewer HDCC treatments compared with placebo. The study drug appeared to be well tolerated.     

Interferon-γ influences the composition of leukocytic infiltrates in murine lyme carditis

Interferon (IFN)-γ is present in lesions of patients with Lyme disease and positively correlates with the severity of manifestations. To investigate the role of IFNγ in the development of Lyme carditis, wild-type and IFNγ-deficient C57BL/6 mice were infected with the causative bacterium, Borrelia burgdorferi. Histological analysis revealed no change in the severity of carditis between wild-type and IFNγ-deficient mice at 14, 21, 25, and 28 days after infection. However, a distinct shift in the types of leukocytes within the hearts of IFNγ-deficient mice was observed at 25 days. In the absence of IFNγ, the number of neutrophils in the heart was increased, whereas the number of T lymphocytes was decreased. Bacterial loads within hearts were the same as in wild-type mice. Macrophages secrete chemokines that recruit immune cells, which could contribute to the accumulation of leukocytes in murine Lyme carditis. The ability of IFNγ and B. burgdorferi to activate murine macrophages was examined, and the two stimuli synergistically induced chemoattractants for mononuclear cells (ie, CXCL9, CXCL10, CXCL11, CXCL16, and CCL12) and decreased those for neutrophils (ie, CXCL1, CXCL2, and CXCL3). IFNγ and B. burgdorferi also synergistically enhanced secretion of CXCL9 and CXCL10 by murine cardiac endothelial cells. These results indicate that IFNγ influences the composition of inflammatory infiltrates in Lyme carditis by promoting the accumulation of leukocytes associated with chronic inflammation and suppressing that of cells that typify acute inflammation.