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Untersuchungen zu Diagnostik und Therapie des Buruli Ulkus mittels tropenadaptierter Labormethoden in endemischen Regionen
Intracapillary leucocyte accumulation as a novel antihaemorrhagic mechanism in acute pancreatitis in mice
Background: Pancreatic infiltration by leucocytes represents a hallmark in acute pancreatitis. Although leucocytes play an active role in the pathophysiology of this disease, the relation between leucocyte activation, microvascular injury and haemorrhage has not been adequately addressed.Methods: We investigated intrapancreatic leucocyte migration, leucocyte extravasation and pancreatic microperfusion in different models of oedematous and necrotising acute pancreatitis in lys-EGFP-ki mice using fluorescent imaging and time-lapse intravital microscopy.Results: In contrast to the current paradigm of leucocyte recruitment, the initial event of leucocyte activation in acute pancreatitis was represented through a dose- and time-dependent occlusion of pancreatic capillaries by intraluminally migrating leucocytes. Intracapillary leucocyte accumulation (ILA) resulted in dense filling of almost all capillaries close to the area of inflammation and preceded transvenular leucocyte extravasation. ILA was also initiated by isolated exposure of the pancreas to interleukin 8 or fMLP, demonstrating the causal role of chemotactic stimuli in the induction of ILA. The onset of intracapillary leucocyte accumulation was strongly inhibited in LFA-1-/- and ICAM-1-/- mice, but not in Mac-1-/- mice. Moreover, prevention of intracapillary leucocyte accumulation led to the development of massive capillary haemorrhages and transformed mild pancreatitis into lethal haemorrhagic disease.Conclusions: ILA represents a novel protective and potentially lifesaving mechanism of haemostasis in acute pancreatitis. This process depends on expression of LFA-1 and ICAM-1 and precedes the classical steps of the leucocyte recruitment cascade.
Interference microscopy delineates cellular proliferations on flat mounted internal limiting membrane specimens.
Aim: To demonstrate that interference microscopy of flat
mounted internal limiting membrane specimens clearly
delineates cellular proliferations at the vitreomacular
Methods: ILM specimens harvested during vitrectomy
were fixed in glutaraldehyde 0.05% and paraformaldehyde
2% for 24 h (pH 7.4). In addition to interference
microscopy, immunocytochemistry using antibodies
against glial fibrillar acidic protein (GFAP) and neurofilament
(NF) was performed. After washing in phosphatebuffered
saline 0.1 M, the specimens were flat-mounted
on glass slides without sectioning, embedding or any
other technique of conventional light microscopy. A cover
slide and 49,6-diamidino-2-phenylindole (DAPI) medium
were added to stain the cell nuclei.
Results: Interference microscopy clearly delineates
cellular proliferations at the ILM. DAPI stained the cell
nuclei. Areas of cellular proliferation can be easily
distinguished from ILM areas without cells.
Immunocytochemistry can be performed without changing
the protocols used in conventional microscopy.
Conclusion: Interference microscopy of flat mounted ILM
specimens gives new insights into the distribution of
cellular proliferations at the vitreomacular interface and
allows for determination of the cell density at the ILM.
Given that the entire ILM peeled is seen en face, the
techniques described offer a more reliable method to
investigate the vitreoretinal interface in terms of cellular
distribution compared with conventional microscopy.
Comparison of CT colonography, colonoscopy, sigmoidoscopy and faecal occult blood tests for the detection of advanced adenoma in an average risk population.
Definite multiple system atrophy in a German family
A variable neurodegenerative phenotype with polymerase gamma mutation