A cytotoxicity assay for the detection and differentiation of two families of shellfish toxins.

There is an urgent need for an alternative to the mouse bioassay for the detection of algal toxins in shellfish on both analytical and animal welfare grounds. Several alternative methodologies have been described, but have not gained widespread acceptance to date, because each assay measures only one or a small number of related phycotoxins out of the increasing range that needs to be detected. A simple cytotoxicity assay using either the HepG2 or ECV-304 cell lines is described with two end-point measurements, which can detect and distinguish between two unrelated classes of phycotoxins. Morphological examination following 3h exposure to the sample enables the detection of the diarrhetic shellfish poisons, including okadaic acid and related toxins. Viability testing using MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide), following 24h exposure of the same cells to the sample, reveals a second class of toxin, which is most probably the newly-described toxin, azaspiracid. This assay should play an important role in shellfish monitoring in the future.

Renal allograft rejection: beta-chemokine involvement in the development of tubulitis.

BACKGROUND: Tubulitis is a defining feature of renal allograft rejection. Graft dysfunction may result from damage inflicted on tubular epithelial cells by intratubular cytotoxic T lymphocytes. Graft cells are known to produce chemokines during acute rejection, but it is not known whether changes in expression of specific chemokines can influence the composition of the intratubular lymphocyte population. We examined expression of individual chemokines in biopsy sections showing different pathological rejection grades. METHODS: Sections from Banff-graded transplant biopsies were examined for the presence of beta-chemokines (MCP-1, MIP-1alpha, MIP-1beta, and RANTES) by immunofluorescence and semiquantitative confocal laser scanning microscopy. RESULTS: Beta-chemokines were expressed predominantly at the basolateral surface of tubular epithelial cells. Expression of MCP-1 and MIP-1beta was significantly higher in sections showing grade 2 rather than grade 1 acute rejection. RANTES and MIP-1alpha showed no significant variation in level of expression between rejection grades. CONCLUSIONS: Beta-chemokines are expressed by tubular epithelial cells during acute rejection. Consistent expression of RANTES and MIP-1alpha suggests a general role in recruiting T lymphocytes. However, MCP-1 and MIP-1beta may play a more subtle role in recruitment of specific T-cell subsets, such as Th1 cells, during acute cellular rejection.

Conversion of renal transplant recipients from cyclosporin to low-dose tacrolimus for refractory rejection.

Twenty-five patients with refractory rejection following renal transplantation were converted from cyclosporin to tacrolimus in an attempt to salvage the allografts. All patients had received two or three pulses of methylprednisolone, 6 had OKT3, 14 had antithymocyte globulin (ATG) and 2 had both OKT3 and ATG prior to conversion. The median time from transplantation to conversion to tacrolimus was 32 days (range 12-322). Patients underwent a simple switch from cyclosporin- to tacrolimusbased therapy with tacrolimus administered at a median dose of 0.15 mg/kg per day. Doses were adjusted according to clinical response and trough blood levels. Twenty-one of the 25 patients (84%) with refractory rejection showed evidence of reversal of rejection as indicated by a significant reduction in serum creatinine (Student's paired t-test, P < 0.05) following conversion to tacrolimus. None of these patients had further episodes of rejection. Three patients had ongoing rejection and returned to dialysis, and 1 patient showed deteriorating renal function associated with a cytomegalovirus infection. Of 18 patients currently on tacrolimus, 15 have improved renal function and 3 have shown no further deterioration. We conclude that low-dose tacrolimus appears to be effective in salvaging renal allografts with resistant rejection.

Conversion of renal transplant recipients from cyclosporin (neoral) to tacrolimus (prograf) for haemolytic uraemic syndrome.

Five patients with cyclosporin-related haemolytic uraemic syndrome (HUS) following cadaveric renal transplantation were converted from cyclosporin- to tacrolimus-based immunosuppression. All patients had biochemical, haematological and biopsy evidence of HUS at the time of conversion. Four of the patients showed complete resolution of the syndrome within 1 week of conversion with normalisation of haemoglobin, platelets and lactate dehydrogenase levels. In the fifth patient renal function stabilised with slow resolution of the haematological and biochemical parameters. Four of the five patients are still taking tacrolimus, one having converted back to cyclosporin due to marked hair loss. We conclude that conversion to tacrolimus appears to be an effective treatment for cyclosporin-related HUS following renal transplantation.

Outcome, survival, and costs in patients undergoing intubation for carcinoma of the esophagus.

BACKGROUND: In this prospective study a consecutive series of 70 patients undergoing insertion of a Wilson-Cook endoprosthesis for palliation of esophageal carcinoma was examined. METHODS: The tube was inserted endoscopically using intravenous sedation and a pulsion technique. RESULTS: The patients had a mean (SEM) age of 70.7 (1.5) years and 44 (63%) were men. Two patients died in hospital and 2 died after discharge, giving a procedure-related mortality of 2.8% and a 30-day mortality of 5.7%. Nine patients experienced complications, giving a morbidity rate of 12.8% following the initial procedure. Twenty patients required a second or further procedure. The indications were tube migration in 22 cases, obstruction in 10, and fistula formation in 2 patients. Thirty-day mortality in this group was significantly greater than after a first procedure (7 patients, 20.1%; P <0.05). The median survival following insertion of a Wilson-Cook endoprosthesis was 16 weeks. CONCLUSIONS: This study describes a safe, effective method for insertion of an endoprosthesis, with a low morbidity and mortality. The average cost for endoscopic insertion of a Wilson-Cook endoprosthesis in this unit is $1,600, and in view of the short median survival in this group of patients, the introduction of costly self-expanding stents is not warranted without demonstrable benefits in a controlled, prospective, randomized clinical trial.

Tumour bed biopsy detects the presence of multifocal disease in patients undergoing breast conservation therapy for primary breast carcinoma.

This study prospectively examined tumour bed biopsies in 135 consecutive patients undergoing conservative surgery for breast carcinoma. All had wide resection of the primary tumour and histologically clear margins. Twelve patients (9%) had positive tumour bed biopsies. Two subgroups of patients had positive bed biopsies; those with ductal carcinoma in situ, and a second group with more aggressive disease characterized by lymph node involvement, vascular invasion and a higher grade and mitotic count. As the majority of recurrences from breast carcinoma occur in the region of the primary tumour, bed biopsy may aid in the identification of a group of patients with multifocal or aggressive disease who are at increased risk of local recurrence.

Developing an information systems strategy for nursing.

With the rapidly changing health care environment and information technology advances, organizations need to engage in strategic, planned change in order to allocate limited resources, achieve the organization's goals, and fulfill its mission [1]. One of the most important aspects of the organization's planned strategies for change concerns the information systems. The involvement of the nursing department in this process is critical. This poster presentation will communicate how nurses can develop an information systems strategic plan that will enable them to play an active role as contributors and vital participants in the strategic and business planning processes for information systems. This information systems strategy for nursing will: a) provide direction and purpose, b) guide nursing in identifying the kinds of information technology needed, c) assist in timely implementation of a system that supports nursing, and d) identify desired outcomes and benefits of an information system. The nursing information systems plan must be built on, and support, the organization's mission and business plan and integrate into the over-all information systems plans [2]. Components of the nursing strategic plan include the nursing mission statement and vision, an assessment of the current environment to identify supporting technology needed to achieve the nursing vision, expectations/anticipated outcomes, environmental considerations, and special staffing/expertise considerations. The nursing vision and mission statement is an articulation of the overall direction and purpose of the nursing organization. An assessment of the nursing organization, problem areas, opportunities for growth, the physical environment, existing systems, communications requirements, and resources is carried out to help identify areas where new technologies and automated methods of managing information could be applied. Special staffing and expertise not currently available in the organization, but necessary to the successful implementation of the plan, should be identified, and plans for filling those needs should be included in the planning and prioritization process. Based on the mission and assessment findings, goals or anticipated outcomes are developed. These goals must be realistic, financially feasible, and logistically achievable; they should also provide direction for action and decision-making [3]. Measurable objectives and detailed action plans can then be developed from these goals when implementation of this aspect of the strategic plan is begun. It is especially important, even at a strategic planning level, to consider change management techniques, including specific steps to involve individuals who will be affected by the change and to ensure open communication throughout the process. Efforts to collaborate with all affected departments and to offer input and educational opportunities to the various members of the health care team should be included in the strategic plan. A business plan describing the mission, goals, and objectives for a specific system implementation is the final step in the strategic planning process. The business plan includes expected outcomes and cost justification and may be done in cooperation with other departments (in the organization) that will be involved with this system. The business plan is used to communicate the information system's needs to the administration and governing board of the organization. With a good information systems strategy, nursing will be prepared to make more timely and better informed decisions related to applying information technology within the nursing department. The end results of this planning should be evident in the improved utilization of information technology to support the nursing vision and mission.

Digestive enzyme levels and histopathology of pancreas disease in farmed Atlantic salmon (Salmo salar).

1. The pancreatic digestive enzyme activities of trypsin and chymotrypsin were assessed in vitro and were found to correlate well with the histological changes characteristic of pancreas disease (PD) in farmed Atlantic salmon (Salmo salar). 2. Pancreatic enzyme activity was assessed in vivo using the chymotrypsin specific substrate N-benzoyl-L-tyrosyl-p-aminobenzoic acid. In all cases, significantly less p-aminobenzoic acid was excreted by fish later found to be suffering from PD. 3. It is concluded that the in vitro digestive enzyme assay was effective in diagnosing PD with the in vivo method, indicating further promise for assessing exocrine pancreatic function.

Differential effects of D600, nifedipine and dantrolene sodium on excitation-secretion coupling and presynaptic beta-adrenoceptor responses in rat atria.

The stimulation-evoked release of tritium was measured from rat atria labelled with [3H]-noradrenaline. The calcium dependence of evoked release and the facilitation of this release via activation of presynaptic beta-adrenoceptors were examined using D600 (methoxyverapamil), nifedipine and dantrolene sodium. Both D600 and nifedipine at dose levels of 20 and 100 microM inhibited evoked release. Dantrolene (20, 100 microM) reduced release by 25%, the effect being maximal at 20 microM. In the presence of 20 nM isoprenaline, a facilitation of evoked release occurred, which was blocked by 0.1 microM (-)-propranolol. The facilitatory action of isoprenaline was abolished by omission of calcium from the buffer, or by D600 or nifedipine, (100 microM). In contrast, the response to isoprenaline was not modified by dantrolene (20, 100 microM). It is concluded that the evoked release of noradrenaline (NA) utilizes Ca from both intra- and extracellular sources and that isoprenaline increases NA secretion by promoting the depolarization-induced influx of Ca.