Detecting autoreactive B cells in the peripheral blood of people with type 1 diabetes using ELISpot.

Type 1 diabetes mellitus (T1D) is an autoimmune disorder where T lymphocytes damage the islet beta cells but B lymphocytes also play an important role. Although changes in peripheral B cell phenotype have been observed, little is known about the B cells that secrete the autoantibodies. We developed a sensitive B cell enzyme-linked immunospot assay (ELISpot assay) to detect individual B cell antibody responses to glutamic acid decarboxylase (GAD) and islet antigen-2 (IA-2). We found that even healthy donors have B cells that secrete antibodies in response to GAD and IA-2 in the ELISpot. There was increased B cell reactivity to autoantigens in the peripheral blood of individuals with newly-diagnosed, but not long-standing, type 1 diabetes. However, no correlation with serum autoantibody levels was found, indicating that additional factors such as antigen affinity or exposure to antigens in vivo are required for antibody secretion, and that even healthy donors have potentially autoreactive B cells.

Timing of completion lymphadenectomy after positive sentinel node biopsy in patients with melanoma.

BACKGROUND: Nodal staging with sentinel node biopsy (SNB) and completion lymph node dissection (CLND) provides prognostic information to patients with melanoma and their physicians. It is not known whether the timing of CLND is associated with survival outcome and/or CLND tumour load. This study investigated whether CLND timing is associated with CLND tumour load, disease-free survival (DFS) and/or melanoma-specific survival (MSS). METHODS: A retrospective cohort of patients with SNB-positive melanoma from nine European Organisation for Research and Treatment of Cancer (EORTC) Melanoma Group centres undergoing surgery between 1993 and 2009 were examined. Patients were selected based on availability of CLND and follow-up data. The CLND interval was defined as the number of days between diagnosis and CLND. Patient and tumour characteristics were collected. Five-year DFS and MSS rates were calculated. Cox and logistic regression analysis were performed, adjusting for known prognostic/predictive indicators. RESULTS: A total of 784 patients were included in the study. Their median age was 51 (i.q.r. 40-62) years, and 418 patients (53·3 per cent) were men. Median Breslow thickness was 3·0 (i.q.r. 2·0-5·0) mm, and 148 patients (18·9 per cent) had a residual tumour load. Median CLND interval was 84 (i.q.r. 65-105) days. Five-year DFS and MSS rates were not significantly different for patients operated on with a median CLND interval of less than 84 days and those with an interval of at least 84 days (DFS: 54·2 versus 53·3 per cent respectively; MSS: 66·9 versus 65·1 per cent). In a multivariable Cox model, CLND interval was not a significant prognostic indicator. CLND interval was negatively correlated with identification of positive non-sentinel nodes, but following adjustment for known risk factors this effect was no longer found. CONCLUSION: The time interval between diagnosis of melanoma and CLND did not influence CLND tumour load, DFS or MSS.

Decision-making influence: an empirical study of social workers in nursing homes.

Health care research on social workers' organizational influence has focused almost exclusively on the hospital setting. Little is known about social workers' influence in nursing homes, in particular organizational factors that support or constrain their impact on resident care. This article presents findings from a survey of 90 social workers employed in 53 nursing homes who identified their level of influence in five spheres of organizational decision making. With the exception of personnel decisions, there were few differences between the influence levels of the directors of social services and social workers without administrative responsibilities. Number of full-time-equivalent social workers, type of ownership, and length of employment of the director of social services were significantly related to the level of perceived influence held by the director of social services. The implications of these findings for social work practice in nursing homes are discussed.

Haemodynamic, pathological, haematological and behavioural changes during endotoxin infusion in equine neonates.

The purpose of this study was to evaluate the effects of experimentally induced sublethal endotoxaemia in equine neonates. Four foals, between two and five days of age, were infused intravenously with 0.5 microgram/kg bodyweight of Salmonella typhimurium endotoxin (LPS) over a 5 h period. A four-day-old and a five-day-old foal, similarly infused with sterile isotonic saline, served as controls. Clinical signs were monitored, blood samples obtained for evaluation of selected haematological and biochemical parameters; and haemodynamic parameters were recorded hourly during the infusion, as well as 6 and 24 h post infusion. Depression, anorexia, increased rectal temperature, leucopenia followed by leucocytosis, hypoglycaemia, increased prothrombin time, partial thromboplastin time (APTT), pulmonary artery pressure, pulmonary artery wedge pressure, right atrial pressure, pulmonary and systemic vascular resistance and mild hypoxaemia were consistent findings in the foals receiving endotoxin. There was marked variation over time in the above parameters, during the infusion. Shock was not induced, and the foals appeared to be healthy shortly after the infusion was discontinued. The return to baseline values of body temperature (3 of 4 foals), APTT (1 of 4 foals) and neutrophil count (2 of 4 foals), during endotoxin infusion, suggests induction of early tolerance. The control foals remained alert and the temperature, prothrombin time and fibrinogen remained stable during the study. Hyperglycaemia, transient increased APTT and variations in selected haemodynamic parameters were recorded in the control foals during the infusion.

Systemic and pulmonary haemodynamics in normal neonatal foals.

Cardiopulmonary function was studied in 10 full-term healthy foals from birth to 14 days of age. Systemic and pulmonary haemodynamics were recorded in lateral recumbency via indwelling aortic and pulmonary artery catheters. Mean body weight increased from 45.4 +/- 2.4 kg on Day 1 to 70.6 +/- 6.1 kg on Day 14. All foals had a continuous murmur of patent ductus arteriosus for 3-6 days. From Day 1 (12 h old) to Day 14, heart rate increased (89 +/- 4 to 95 +/- 5/min), mean aortic pressure increased (87.7 +/- 1.9 to 100.3 +/- 3.2 mmHg), mean pulmonary artery pressure decreased (38.6 +/- 4.6 to 27.4 +/- 3.0 mmHg), mean right atrial pressure was unchanged (4.5 +/- 0.5 to 4.6 +/- 0.9 mmHg), mean pulmonary artery wedge pressure was unchanged (7.6 +/- 0.9 mmHg to 8.1 +/- 0.7 mmHg), cardiac output increased (8.03 +/- 0.59 to 15.88 +/- 1.90 l/min), cardiac index increased (180.5 +/- 10.3 to 222.1 +/- 21.6 ml/kg/min), stroke volume increased (90.4 +/- 5.7 to 164.2 +/- 25.9 ml), stroke volume index was unchanged (2.04 +/- 0.10 to 2.30 +/- 0.35 ml/kg), pulmonary vascular resistance decreased (314 +/- 39 to 104 +/- 21 aru), systemic vascular resistance decreased (858 +/- 70 to 497 +/- 87 aru), and pulmonary/systemic resistance ratio decreased (38 +/- 6 to 21 +/- 5%). All changes were gradual, although pulmonary artery pressure and pulmonary vascular resistance decreased rapidly in the first 24 h. Catheters were well tolerated over several days, indicating their feasibility for studying cardiovascular function in full-term or premature equine neonates.